Hexarelin, Ghrelin, GHRP-6/2

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    Hexarelin, Ghrelin, GHRP-6/2


    Lately I have been doing some casual research on the use of these peptides for increasing GH. As of now, these are all legal for research purposes (Hex and GHRP-6 being the actives in IBE's Hexatropin). Does anybody have any experience with these peptides as an injectible? Have any of you clever fellas (you know who you are) come across any information regarding this? Thoughts, Feelings?

    Here are some of the key things I have found:

    -Research has shown that all three Compounds substantially increase HGH production in human subjects between the ages of 30-45 years old. (older subjects required actual HGH replacement)
    -The degree of increase is dependent on the subjects natural GHRH production (which explains the age thing above)
    -Dosages used in research are around 2mcg per pound of lean body weight.
    -At these dosages, Ghrelin and GHRP-2 are pretty inexpensive (Don't know if I'm allowed to post the prices I found, so I'm gonna err on the side of caution)
    -GHRP-6 has been shown to increase appetite signifigantly. GHRP-2 is the same compound without the appetite increase.


    Sorry for the incomplete nature of this post, it was kinda thrown together...I will post some links and hopefully more information later, if anyone is interested. Just wanted to get some thoughts.

    Looking forward to some feedback.....

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    hmm, good work. do you mind emailing me? i have a comment i want to run past you before i post it.

    reeses@safe-mail.net
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    Thanks for the reply...e-mail sent brotha. Looking forward to some input.
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    Go ahead and post prices. I am interested. I will take the blame if anything is said, which I dont think it will be.
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    Hey LMD,

    GHRP-6 seems to be around 1mg/$50...the rest are a little more expensive, but with the GHRP being so cheap I was thinking about a little ****tail, I just need more research to work out the details.
    Hexarelin is 1mg/$165 (@2mcg/#of lbm could get pretty expensive)
    Ghrelin is .1mg/$90-$100 (ouch)

    (Mods...please delete if this is inapropriate, I wasn't sure)
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    Quote Originally Posted by myfathersboy
    Hey LMD,

    GHRP-6 seems to be around 1mg/$50...the rest are a little more expensive, but with the GHRP being so cheap I was thinking about a little ****tail, I just need more research to work out the details.
    Hexarelin is 1mg/$165 (@2mcg/#of lbm could get pretty expensive)
    Ghrelin is .1mg/$90-$100 (ouch)

    (Mods...please delete if this is inapropriate, I wasn't sure)

    Seems a lil too expensive for my taste lol.
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    Actually....um, heres a little edit to the original post:
    2mcg/kilogram of LBM....sorry, my mistake.

    LMD, that's not really that bad if you followed a 5on/2off dosing protocol, depending on your lean body weight (for the GHRP-6 anyway). As for the Hex... yeah, a little pricey, which actually kinda makes me wonder how IBE is able to sell Hexatropin (containing 2mg total Hexarelin) for $130...they must be getting some good pricing on the stuff. Also there is a good possibility that lower doses would be effective too(once again, to use Hexatropin as an example the dosages are 100mcg of each per dose...then again there is the mysterious cell mediated carrier to factor in). And hell, It's infinately less than HGH. Maybe I'll see if IBE can source these items for a more managable price (but only for research purposes, of course).
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    Quote Originally Posted by myfathersboy
    Actually....um, heres a little edit to the original post:
    2mcg/kilogram of LBM....sorry, my mistake.

    LMD, that's not really that bad if you followed a 5on/2off dosing protocol, depending on your lean body weight (for the GHRP-6 anyway). As for the Hex... yeah, a little pricey, which actually kinda makes me wonder how IBE is able to sell Hexatropin (containing 2mg total Hexarelin) for $130...they must be getting some good pricing on the stuff. Also there is a good possibility that lower doses would be effective too(once again, to use Hexatropin as an example the dosages are 100mcg of each per dose...then again there is the mysterious cell mediated carrier to factor in). And hell, It's infinately less than HGH. Maybe I'll see if IBE can source these items for a more managable price (but only for research purposes, of course).
    Guess I can start a new research quest and be an expert in another field haha.

    Yeah they get good prices on stuff from China. Large bulk quantities of coures..........for research purposes only.
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    Quote Originally Posted by LakeMountD
    Guess I can start a new research quest and be an expert in another field haha
    Well, I'm gonna try to shed some light on this one...but as far as research and being an 'expert' go...you've set the bar pretty high with all that MGF/IGF-1 info Not sure if I can even come close (where do you find the time anyway??) Of course any contributions would, as always, be appreciated.
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    Quote Originally Posted by myfathersboy
    Lately I have been doing some casual research on the use of these peptides for increasing GH.

    Here are some of the key things I have found:

    -Research has shown that all three Compounds substantially increase HGH production in human subjects between the ages of 30-45 years old. (older subjects required actual HGH replacement)
    -The degree of increase is dependent on the subjects natural GHRH production (which explains the age thing above)
    Have not seen anything to indicate GHRP non-function in subjects over 45; studies out of Torino (U. Turin) found the opposite IIRC.

    My own experience has been pretty compelling so far: indicators reflect increased GH output.
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    BW,

    I appreciate your input, your log is an excellent read and very inspirational. The point I was trying to make (albeit, it could have been a bit clearer) is that researchers have found that subjects over 45 have more pronounced results from HGH than from any of the GH releasing peptides, which is true, I imagine, of subjects at any age...just that the difference is more pronounced. Did that make sense? I wasn't insinuating(nor was the study, that I can tell) that these compounds aren't effective for certain age groups.

    On a different note, do you have any other thoughts on this?
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    Thanks IBE!!! Please keep us posted on your progress in getting these items...I always have great experiences when dealing with you guys, and quite frankly I am very interested in experimenting with these compounds and would rather buy from a known and trusted entity.

    What do you think about dosages?

    Anything else to contribute?

    Thanks again....

    Oh...and because I am pretty sure someone will mention it...yes I'm planning to do a log.
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    Quote Originally Posted by IBE
    that company that sell's it for 165 per 1mg is a rip off, I know what company you are talking about. hexarelin is now hard to find do to one chem needed...2methyl which alot of companys stopped making 2methyl. we get all our peptides from USA. we will be selling the injectable GHRP-6 again soon and also inject. hexarelin (hopefully)
    cool. i'd be interested. what about GHRP-2?
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    IBE sent me hexatropin and injectible GHRP-6 to compare. I ran the oral first with good resutls (note I'm an experienced GH user). when I came to reconsitute the GHRP it was empty. IE nothing was inside the ampule. I still have it in my fridge and can upload a pic if need be.
    I forgot all about it until I saw this thread. basically I said at the times I liked the thought of injecting GHRP better but IBE said the oral is longer acting and offerend a trial. sorry I couln't complete the trial, the oral stuff is good **** though and I do wonder how the injectible will rank up to it.
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    Quote Originally Posted by judge-mental
    IBE sent me hexatropin and injectible GHRP-6 to compare. I ran the oral first with good resutls (note I'm an experienced GH user). when I came to reconsitute the GHRP it was empty. IE nothing was inside the ampule. I still have it in my fridge and can upload a pic if need be.
    I forgot all about it until I saw this thread. basically I said at the times I liked the thought of injecting GHRP better but IBE said the oral is longer acting and offerend a trial. sorry I couln't complete the trial, the oral stuff is good **** though and I do wonder how the injectible will rank up to it.

    I imagine the vial was not empty....some peptides are...umm, well **** I don't really have the vocabulary to describe it but...The peptide itself is so insubstantial that it is just sprayed into the vial and clings to the glass...it's in there, but nearly invisible to the naked eye. look at your vial again and see if you notice a slight 'fog' on the inside of the vial. I really can't imagine IBE sending you an empty vial...then again, stranger things have happened.

    I too am interested in the comparison between the cell mediated oral and the injectible, cost and effectiveness...but especially continued availability, as these compounds really need continued use to realize maximum results.

    Thanks for contributing, and let us know about that vial, if you still have it.
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    Just to chime in... I have been running Hex-6 for 3 1/2 months and it ROCKS!

    Plan to start a 3 month Ecto run soon
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    very intresting
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    Quote Originally Posted by EEmain
    Just to chime in... I have been running Hex-6 for 3 1/2 months and it ROCKS!

    Plan to start a 3 month Ecto run soon
    Thanks EE, did you do a log?

    I am hoping that the injectible versions of these compounds will be a little more cost effective for running long-term, and possibly show even more pronounced effect than the cell-mediated oral.
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    Quote Originally Posted by myfathersboy
    Thanks EE, did you do a log?

    I am hoping that the injectible versions of these compounds will be a little more cost effective for running long-term, and possibly show even more pronounced effect than the cell-mediated oral.
    Not a paperwork kinda guy Not! just lazy I plan on keeping a log of the Ecto cut however.
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    Quote Originally Posted by EEmain
    Not a paperwork kinda guy Not! just lazy I plan on keeping a log of the Ecto cut however.
    looks like you got a nice deal on that ecto!!!
    Looking forward to your cutting log.....
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    Quote Originally Posted by EEmain
    Just to chime in... I have been running Hex-6 for 3 1/2 months and it ROCKS!

    Plan to start a 3 month Ecto run soon
    how so? what effects have you noticed? what else are you running?
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    Quote Originally Posted by myfathersboy
    looks like you got a nice deal on that ecto!!!
    Looking forward to your cutting log.....
    And the CissusRx too
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    Quote Originally Posted by same_old
    how so? what effects have you noticed? what else are you running?

    First 2 months just Hex... besides all of BW observations in his log I (and others) have seen a noticable increase in muscle size along with a loss in bodyfat. However my weight has remained steady. That is after I firgured out my salt intake was causing fluctuations. Deep dreams, vivid almost like awakened dreaming. Lethargy only when complex carbs were low. The higher the better... spread through the day. Constant SWOLE feeling and pumps from hell at times.

    Added Mohn and my strength and muscle hardness took on new meaning.
    Messed up my Mohn dosing schedule but haven`t really seen any fall back yet. 4 more days of Mohn at 32mg and 10on of Hex 5on-2off.

    Something else I never mentioned before. I`ve been taking 2 GXR with PWO shake ( 2 cups oats/ 2 scoops WPI) throughout.
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    Quote Originally Posted by myfathersboy
    Turkey Burgers ummmmm
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    Quote Originally Posted by EEmain
    First 2 months just Hex... besides all of BW observations in his log I (and others) have seen a noticable increase in muscle size along with a loss in bodyfat. However my weight has remained steady.

    Added Mohn and my strength and muscle hardness took on new meaning.
    Messed up my Mohn dosing schedule but haven`t really seen any fall back yet. 4 more days of Mohn at 32mg and 10on of Hex 5on-2off.
    have you measured anything? taken BF%? what were you trying to accomplish with hex?
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    Quote Originally Posted by myfathersboy
    I imagine the vial was not empty....some peptides are...umm, well **** I don't really have the vocabulary to describe it but...The peptide itself is so insubstantial that it is just sprayed into the vial and clings to the glass...it's in there, but nearly invisible to the naked eye. look at your vial again and see if you notice a slight 'fog' on the inside of the vial. I really can't imagine IBE sending you an empty vial...then again, stranger things have happened.

    I too am interested in the comparison between the cell mediated oral and the injectible, cost and effectiveness...but especially continued availability, as these compounds really need continued use to realize maximum results.

    Thanks for contributing, and let us know about that vial, if you still have it.
    I will take a close up photo and upload it.
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    Here's the pic of the vial. Note that I shot IGF and GH before and know how tiny doses look like.

    Sorry for the condition of the label, it was in a bag with a transdermal that spilled. You can still see the label though.
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    Quote Originally Posted by same_old
    have you measured anything? taken BF%? what were you trying to accomplish with hex?
    No. I don`t give a **** about numbers. Not being ignorant I just don`t care. Bodyweight and lifts plus visuals give me all I need to know.

    What am I trying to accomplish? The fountain of youth Seriously Hgh interested me but I am pinaphobic, always have been and probably will be. So it came down to PGHT or Hex. And Hex won. My goals are that I started 2 years ago at nearly 300lbs at 30%bf (see attached) and have been working back into shape to improve my lifestyle and maybe extend my stay on this rock. When I am through with all this maybe PGHT is in the future. Who knows?
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    Quote Originally Posted by judge-mental
    Here's the pic of the vial. Note that I shot IGF and GH before and know how tiny doses look like.

    Sorry for the condition of the label, it was in a bag with a transdermal that spilled. You can still see the label though.
    Sorry bro, I wasn't insinuating any lack of experience....but some (dare I say many?) peptides are nearly invisible to the naked eye...literally 'sprayed' into the vial...not like IGF-1 where you can actually SEE 1mg of crystal in the bottom of the vial...I was just wondering if this was the case with GHRP-6 (as I have never actually seen it)....That being said, that vial does look pretty empty doesn't it?????
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    Quote Originally Posted by IBE
    if you are referring to the GHRP-6 vial? this comes fromm a big peptide company so i think they would of sent us an empty vial. do you still have the unopened vial?
    Did you mean to say that you do or don't think they would have sent you an empty vial? (the sentence was just worded kind of strange...)

    I see on the site (nice new digs, by the way) that you have GHRP-6 listed as 'coming soon'....any word on the hexarelin? and on behalf of Same old, how about the GHRP-2?

    Thanks for your responses in this thread, you have been extremely helpful.
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    GHRP-6 now available at IBE.....So

    Bump to this ?????
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    Quote Originally Posted by myfathersboy
    Actually....um, heres a little edit to the original post:
    2mcg/kilogram of LBM....sorry, my mistake.

    LMD, that's not really that bad if you followed a 5on/2off dosing protocol, depending on your lean body weight (for the GHRP-6 anyway). As for the Hex... yeah, a little pricey, which actually kinda makes me wonder how IBE is able to sell Hexatropin (containing 2mg total Hexarelin) for $130...they must be getting some good pricing on the stuff. Also there is a good possibility that lower doses would be effective too(once again, to use Hexatropin as an example the dosages are 100mcg of each per dose...then again there is the mysterious cell mediated carrier to factor in). And hell, It's infinately less than HGH. Maybe I'll see if IBE can source these items for a more managable price (but only for research purposes, of course).
    Bro, if you got the right connections and the necessary credentials you can buy GHRP-6 for as low as $1/MG, $2/GHRP-2MG and $4/MG of Hexarelin..those are the prices i get those anyways. I am sure IBE gets good prices as well. Those who knwo me here know what i do and I am not bsing and those who don't I am sorry I meant no disrespect but since he posted the prices I thought I post the real prices outhere..sure even my supplier's catalog offers GHRP-6 for like $30/MG but when you buy BULK you get huge discounts and I mean huge!

    Thanks,

    Carlito
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    Quote Originally Posted by myfathersboy
    Did you mean to say that you do or don't think they would have sent you an empty vial? (the sentence was just worded kind of strange...)

    I see on the site (nice new digs, by the way) that you have GHRP-6 listed as 'coming soon'....any word on the hexarelin? and on behalf of Same old, how about the GHRP-2?

    Thanks for your responses in this thread, you have been extremely helpful.
    Please make no mistake to note that not all lyophilized peptides look just like IGF-1LR3! I just received 500 vials of MGF and each one came in small plastic vials with screw on caps and each had 2mgs of MGF,m these were almost half the size of the regular IGF-1 you see ( i do know that IGF-1 has something else added to ) at first I was a bit worried as i expected more powder volume...anyways, I pour the lyophilized powder which looks like a large rice grain and a bit solid sort of like a spider's eggs...so I pout it into an empty vial and crimped it then added 2mls of STERILE WATER and it dissolved pretty fast...been using it for 3 days now and i see some results . I am using it by itself and 200mcgs daily.

    Later,

    Carlito
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    Quote Originally Posted by Carlito
    Bro, if you got the right connections and the necessary credentials you can buy GHRP-6 for as low as $1/MG, $2/GHRP-2MG and $4/MG of Hexarelin..those are the prices i get those anyways. I am sure IBE gets good prices as well. Those who knwo me here know what i do and I am not bsing and those who don't I am sorry I meant no disrespect but since he posted the prices I thought I post the real prices outhere..sure even my supplier's catalog offers GHRP-6 for like $30/MG but when you buy BULK you get huge discounts and I mean huge!

    Thanks,

    Carlito
    I have read some of your posts in the past....so I sort of have a vague idea of what it is you do, thanks for posting, your comments are appreciated....

    Hex for $4???.....Seriously not doubting you but, there must be some kind of huge catch to get these prices, I have done quite a bit of searching for information on these peptides and the lowest price I have found on Hex is like $135/mg!!!! and GHRP-6 and 2 for around $35.....What kind of volume and credentials are we talking about here?
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    Thank you,

    What I did before is now in the past though, no more of that!
    My current business is legitimate and I have it registered as well and yes I meant $4/mg but of course if I buy it by the gram...not to mention packaging and other stuff.

    I have website now and more but for obvious reasons I can't post, however I am doing it in my homeboard OL under a new name...all i can say about that.

    Thanks for your reply

    Carlito



    Quote Originally Posted by myfathersboy
    I have read some of your posts in the past....so I sort of have a vague idea of what it is you do, thanks for posting, your comments are appreciated....

    Hex for $4???.....Seriously not doubting you but, there must be some kind of huge catch to get these prices, I have done quite a bit of searching for information on these peptides and the lowest price I have found on Hex is like $135/mg!!!! and GHRP-6 and 2 for around $35.....What kind of volume and credentials are we talking about here?
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    yes,igf has manitol as excipient material for lyophilization all that powder is not pure igf.it ranges from 400-800 or so mg total volume in a 1mg lr3 vial

    please keep us posted on your hex experiment.what are the main benefits supposed to be of that?

    Quote Originally Posted by Carlito
    Please make no mistake to note that not all lyophilized peptides look just like IGF-1LR3! I just received 500 vials of MGF and each one came in small plastic vials with screw on caps and each had 2mgs of MGF,m these were almost half the size of the regular IGF-1 you see ( i do know that IGF-1 has something else added to ) at first I was a bit worried as i expected more powder volume...anyways, I pour the lyophilized powder which looks like a large rice grain and a bit solid sort of like a spider's eggs...so I pout it into an empty vial and crimped it then added 2mls of STERILE WATER and it dissolved pretty fast...been using it for 3 days now and i see some results . I am using it by itself and 200mcgs daily.

    Later,

    Carlito
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    Quote Originally Posted by Carlito
    Thank you,

    What I did before is now in the past though, no more of that!
    My current business is legitimate and I have it registered as well and yes I meant $4/mg but of course if I buy it by the gram...not to mention packaging and other stuff.

    I have website now and more but for obvious reasons I can't post, however I am doing it in my homeboard OL under a new name...all i can say about that.

    Thanks for your reply

    Carlito
    Carlito, you will be an asset to the community for sure, but you REALLY need to become a paying sponsor on a non-source board like this one. you're stepping on some big toes in here.
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    Quote Originally Posted by same_old
    Carlito, you will be an asset to the community for sure, but you REALLY need to become a paying sponsor on a non-source board like this one. you're stepping on some big toes in here.
    I understand your point and I have tried before and Bobo said he had no room for me so...

    One of my products is already advertise in 30 boards thru a friend's new research company and we are moving hundreds of units monthly...soon he'll carry MGF , GHRP-6, GHRP-2, Hexarelin, Sermorelin Acetate and much more to come...

    I do not mean no disrespect so I will never mention my business here again.

    Thank you,

    Carlito
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    Quote Originally Posted by IBE
    Your MGF will not sell once we come out with the PEGylated version. and for GHRP-6 they all really need to be in a delivery system to be most effective because of there short half life. there is not point selling GHRP-2 because it is no more potent than GHRP-6 or hexarelin so it will be a waste of time.
    SUre bro

    Thanks for the LOVE

    The MGF is working fine on me and though I am using it by itself I've noticed strength increase ( slight ) but mainly I've noticed my metabolism has increased and also more stamina, I am on nothing else and using 400mcgs ( now ) daily divided in 4 injections...injecting is not a problem as i dilluted my 2mgs in one ml od sterile water so each 10 mark on a 1cc slinpin is 200mcgs so that is a very small amount to inject and it makes multiple injects a breeze.

    Lyophilized GHRP-6 dilluted in sterile water ( shelf life of one month once dilluted under 32 degrees ) and ready to be injected sub q . Injections are the way to go and most know that!
    Oral delivery though ideal is just not the best choice and YOU KNOW IT bud.

    Later !

    Carlito

    PS: SERMORELIN ACETATE
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    Commentary
    Signal transduction systems employed by synthetic GH-releasing
    peptides in somatotrophs
    C Chen, D Wu and I J Clarke
    Introduction
    Around the time when the endogenous hypothalamic
    releasing factor for growth hormone (GH) was isolated, a
    new range of synthetic peptides were also shown to be
    specific releasors of GH. These peptides were originally
    developed in the late 1970s by Bowers et al. (1980) who
    synthesized the enkephalin analogue Tyr-D-Trp-Gly-
    Phe-Met-NH2 and found that it specifically stimulated
    GH secretion. Although this pentapeptide was a weak GH
    secretagogue it was the first non-GH-releasing hormone
    (GHRH) found to act on the anterior pituitary to specifi-
    cally release GH and was used as a model to design
    more potent forms of GH-releasing peptides (GHRPs)
    (Momany et al. 1981). The most potent of the first
    generation analogues was GHRP-6 (Bowers et al. 1984,
    Momany et al. 1984), a hexapeptide which stimulates GH
    release in a variety of animal species and in man (Bowers
    et al. 1984, 1991, Wu et al. 1994a). Since then, a range of
    compounds has been developed with varying potency
    including GHRP-1 and GHRP-2. In spite of their
    obvious eYcacy and specificity, no endogenous homologue
    has been found for these synthetic peptides and the
    receptor through which they exert their eVect has not
    been identified. Nevertheless, some progress has been
    made in defining the signal transduction systems that
    mediate responses to GHRPs. Some of the peptides act in
    a synergistic or additive way with endogenous GHRH and
    this has provided a basis for their clinical use. Since the
    GHRPs are short peptides (5–7 amino acid residues), they
    are synthesized easily and are not as readily degraded in
    plasma as GHRH; these features ameliorate their clinical
    potential. Because of their chemical nature the GHRPs are
    eYcacious when administered orally or intravenously
    (Bowers et al. 1990, 1992).
    If endogenous GHRPs do exist then how could they be
    rationally integrated into the picture for the control of GH
    that has been developed on the basis that GHRH stimulates
    and somatostatin (SRIF) inhibits secretion (Brazeau
    et al. 1973, Rivier et al. 1982, Guillemin et al. 1982)? A
    regulatory model has been established in the rat which
    dictates that GH release is the sum of the combined eVects
    of GHRH and SRIF (Tannenbaum & Ling 1984). There
    is some experimental evidence for this, since reciprocal
    alterations in the hypophysial portal blood levels of GHRH
    and SRIF seem to exist (Plotsky & Vale 1985) and relate
    to the pulsatile secretion of GH. On the other hand,
    studies in sheep (Frohman et al. 1990, Thomas et al. 1991,
    Magnan et al. 1994) do not indicate such a close relationship
    between GHRH, SRIF and GH, calling into question
    the general applicability of the rat model. Perhaps
    there is a role for an endogenous GHRP in the regulation
    of GH secretion in sheep and other species.
    Given the range of GHRPs that have now been
    synthesized and studied, it seems timely to review what is
    known about their mechanism of action. An excellent
    recent review by Korbonits & Grossman (1995) discussed
    in vivo and in vitro actions of GHRPs on GH secretion and
    the possible clinical application of GHRPs. The present
    article focusses on the intracellular signal transduction
    systems in somatotrophs employed by GHRPs with a brief
    overview of the site of action, chemical structure and
    possible physiological roles.
    Site of action of GHRPs
    It is likely that GHRPs stimulate GH release by both
    direct action on pituitary somatotrophs and hypothalamus
    (Bowers et al. 1991, Fletcher et al. 1994, Guillaume et al.
    1994, Fletcher et al. 1995).
    Hypothalamic action GHRPs do not appear to inhibit
    SRIF release (Hao et al. 1988, Guillaume et al. 1994,
    Korbonits & Grossman 1995). The issue of whether or not
    these peptides have any hypothalamic action on GHRH or
    other factors is a little unclear. GHRP-6 has been shown
    to activate neurones of the arcuate nucleus that project to
    the median eminence (****son et al. 1995), but infusion in
    the third ventricle had a minor eVect on the pulsatile
    release of GHRH in conscious ewes and no eVect on SRIF
    secretion (Fletcher et al. 1995). Moreover, GHRPs did not
    acutely stimulate GHRH release with perifusion of rat
    hypothalamus in vitro (Korbonits & Grossman 1995).
    381
    Journal of Endocrinology (1996) 148, 381–386 ? 1996 Journal of Endocrinology Ltd Printed in Great Britain
    0022–0795/96/0148–0381 $08.00/0
    Hexarelin, a more potent peptide, elevated GHRH levels
    in the hypophysial portal blood of rams (Guillaume et al.
    1994) with no eVect on SRIF secretion. GHRPs are able
    to potentiate GH release in response to a maximally
    stimulating dose of GHRH (Bowers et al. 1990) and
    anti-GHRH antibody infusion cannot abolish the eVect
    of GHRP-6 on GH release from rat pituitary glands
    (Locatelli et al. 1994). These latter observations suggest
    action at the pituitary level or, possibly, through a non-
    GHRH hypothalamic factor yet to be identified (Bowers
    et al. 1990).
    Pituitary action All GHRPs act directly on the pituitary
    gland in vitro (Akman et al. 1993, Bowers 1993, Cheng
    et al. 1993, Bowers et al. 1994, Wu et al. 1994a,b, Chen &
    Clarke 1995b) and in vivo (rat; Mallo et al. 1993, sheep;
    Fletcher et al. 1994) to increase GH secretion. Long-term
    (days) GHRP-6 treatment of GHRH-deprived infant rats
    stimulated GH gene expression which counteracted the
    GHRH deprivation (Locatelli et al. 1994). This action of
    GHRP-6 on GH synthesis is, however, abolished when
    the pituitary is anatomically disconnected from the hypothalamus
    which suggests an involvement of unknown
    hypothalamic factors. The eVect of GHRPs on GH
    synthesis requires further study before any conclusions can
    be drawn.
    Structure and potency of GHRPs
    Since the development of the first peptide in the GHRP
    series (GHRP-6), a range of more potent compounds
    including non-peptidergic GH secretagogues have been
    developed (Deghenghi et al. 1992, Akman et al. 1993,
    Bowers 1993, Smith et al. 1993). Because it has been
    demonstrated that non-peptidergic GH secretagogues act
    in a similar way to GHRP-6 (Cheng et al. 1993, Smith
    et al. 1993), these non-peptidergic compounds are not
    separately considered below.
    The structure and relative potencies of the most wellcharacterized
    of the GHRPs is given in Table 1. GHRP-2
    is the most potent, being equivalent to GHRH in terms of
    its ability to stimulate GH release from sheep pituitary cells
    in vitro (Wu et al. 1994b). GHRP-1 has been reported to
    release GH in the rat with the same potency as GHRH
    (Akman et al. 1993), but this is not true for ovine cells (Wu
    et al. 1994a). GHRP-6 was found to stimulate GH
    secretion in most human acromegalic tumours in vitro
    whereas GHRH stimulated GH release in only some
    (Renner et al. 1994). A mutation in the subunit of the Gs
    protein has been reported in a proportion of such tumours
    as being responsible for constitutive activation of adenylyl
    cyclase activity in response to GHRH (Harris et al. 1992);
    this may explain the latter finding. The adenylyl cyclase
    pathway may not necessarily be stimulated by GHRPs (see
    below), whereas this is a recognized second messenger
    pathway for GHRH (Harwood et al. 1984).
    Intracellular second messenger systems employed by GHRPs
    Intracellular free Ca2+ and membrane ion channels
    GH secretion is directly related to the intracellular free
    calcium concentration ([Ca2+]i) (Lussier et al. 1991). Ca2+
    influx through voltage-dependent Ca2+ channels is stimulated
    by GHRH and reduced by SRIF (Chen et al.
    1989a,b, 1990, 1992, Lussier et al. 1991, Chen & Clarke
    1995a). There is no clear evidence, however, that GHRH
    or SRIF mobilizes Ca2+ from intracellular Ca2+ storage
    sites. On the other hand, GHRP-6 causes the release of
    intracellular Ca2+ as well as Ca2+ influx through the
    cell membrane (Bresson-Bepoldin & Dufy-Barbe 1994,
    Herrington & Hille 1994). In isolated rat somatotrophs,
    GHRP-6 evoked dual-phase increases in [Ca2+]i; an
    initial phase transient increase in [Ca2+]i due to Ca2+
    release which was not blocked by the Ca2+ channel
    blocker, and a second long-lasting phase that was due
    to Ca2+ influx (Bresson-Bepoldin & Dufy-Barbe 1994,
    Herrington & Hille 1994). In ovine pituitary cells,
    GHRP-1 caused a subtle and transient increase in [Ca2+]i
    even when extracellular Ca2+ was chelated to zero (K
    Katoh, C Chen, D Wu, J Zhang, I J Clarke, C Y Bowers
    & D Engler, unpublished observations). This probably
    involves the generation of inositol trisphosphate (InsP3)
    (Adams et al. 1995, Lei et al. 1995) but this has not yet
    been proven. In spite of the mobilization of Ca2+ from
    intracellular stores, the major contribution to the elevation
    of [Ca2+]i is caused by Ca2+ influx via Ca2+ channels. It
    would appear that this is an integral factor in the release of
    GH in response to GHRPs because Ca2+ channel blockade
    abolishes secretion (Akman et al. 1993, Wu et al.
    1994a,b).
    In somatotrophs, the major Ca2+ channels are the
    voltage-gated T- and L-types (Chen et al. 1990, Chen &
    Clarke 1995a). Studies of rat and sheep cells have respectively
    shown that GHRP-6 and GHRP-2 depolarize the
    cell membrane potential leading to the opening of these
    channels (Herrington & Hille 1994, Chen & Clarke
    1995b). Since this depolarization can only be recorded
    with the nystatin-perforated patch clamp configuration
    Table 1 Chemical structure and the relative potency of GHRPs in
    terms of ability to stimulate GH release. Potency is presented in
    relation to the potency of GHRH (unity) in rat and ovine pituitary
    cells in vitro
    Chemical structure
    In vitro
    potency
    (relative to
    GHRH)
    Name
    GHRP-6 His-D-Trp-Ala-Trp-D-Phe-Lys-NH2 0·01–0·1
    GHRP-1 Ala-His-D-‚Nal-Ala-Trp-D-Phe-Lys-NH2 0·1–1
    GHRP-2 D-Ala-D-‚Nal-Ala-Trp-D-Phe-Lys-NH2 1

    Commentary 382
    Journal of Endocrinology (1996) 148, 381–386
    (which does not disturb intracellular second messenger
    systems), this implies that an intact intracellular second
    messenger signalling system is required. In addition, recent
    data indicate that GHRP-2 acts on voltage-gated Ca2+
    channels via second messenger systems, leading to an
    increase in transmembrane L- and T-type Ca2+ currents
    (Chen & Clarke 1995b). All these eVects of GHRPs on
    electrophysiological properties of the membrane of somatotrophs
    resemble the eVects of GHRH but are opposite to
    the eVects of SRIF.
    The ion channels that are involved in depolarization of
    the somatotroph cell membrane are not defined. Studies to
    date show that depolarization caused by GHRPs does not
    involve the Na+ channel in a major way (Herrington &
    Hille 1994, Chen & Clarke 1995b). It is thought that the
    voltage-gated Ca2+ channel activation is partly responsible
    for the depolarization caused by GHRP-2 (Chen & Clarke
    1995b). K+ channels may also be involved but these have
    not been fully characterized (Pong et al. 1991, Chen &
    Clarke 1995b).
    In summary, it is proposed that GHRPs first releases
    intracellular Ca2+ and then causes Ca2+ influx by an
    increase in membrane Ca2+ permeability. The latter is due
    to membrane depolarization and the action of second
    messengers on Ca2+ channel proteins. Possible second
    messenger pathways involved in the action of GHRPs are
    discussed in the following sections.
    The cyclic AMP (cAMP)/protein kinase A (PKA)
    pathway It is well established that GHRH activates the
    cAMP/PKA pathway in somatotrophs and this is fundamental
    to the release of GH (Harwood et al. 1984,
    Frohman et al. 1992). Part of the eVect of SRIF is via its
    inhibition on cAMP formation (Schonbrunn 1990). In
    contrast, GHRP-6 and GHRP-1 have no direct eVect on
    intracellular cAMP levels in rat and ovine somatotrophs
    (Cheng et al. 1989, Akman et al. 1993, Wu et al. 1996).
    Nevertheless, GHRP-6 may synergize with GHRH to
    increase intracellular cAMP levels (Cheng et al. 1989).
    Similar results were also found for the non-peptidergic
    analogue L-692,429 (Cheng et al. 1993), but synergy
    could not be demonstrated with GHRP-1 and GHRH
    (Akman et al. 1993). This discrepancy suggests that an
    increase in cAMP levels is not the primary signalling
    pathway for GHRP-6 and GHRP-1.
    GHRP-2 increased intracellular cAMP levels in ovine
    (but not rat) somatotrophs (Wu et al. 1996). GHRP-2-
    stimulated GH secretion was also blocked in these cells by
    an inhibitor of adenylyl cyclase and a cAMP-binding
    antagonist. Thus, in sheep cells at least, GHRP-2 activates
    adenylyl cyclase leading to an increase in cAMP levels and
    activation of cAMP-dependent PKA. Thus, PKA could
    phosphorylate transmembrane Ca2+ channels to modify
    their properties in the manner observed by electrophysiological
    studies (see above); further work is needed to
    clarify this point. A significant species diVerence is seen
    in sheep and rat somatotrophs in terms of GH release in
    response to GHRP-2 (Bowers 1993, Wu et al. 1994b).
    The increase in adenylyl cyclase activity is obtained in
    ovine pituitary cells with GHRP-2 but not GHRP-6 or
    GHRP-1 (Wu et al. 1996). We propose therefore that, in
    ovine somatotrophs, the pathway employed by GHRP-2,
    resulting in an increase in cAMP, is diVerent from that
    employed by GHRP-6 or GHRP-1 in rat or ovine
    pituitary cells (Cheng et al. 1989, 1993, Akman et al. 1993,
    Wu et al. 1994a).
    The mechanism of activation of adenylyl cyclase by
    GHRP-2 is not clear. Although some subtypes of adenylyl
    cyclase can be activated by Ca2+ (Cooper et al. 1995), the
    subtype of adenylyl cyclase that is involved in the response
    to GHRP-2 may not be stimulated by Ca2+ because
    blockade of Ca2+ influx did not aVect the increase in
    cAMP levels in response to GHRP-2 (Wu et al. 1996).
    Although it is clear that GHRH elevates cAMP levels in
    ovine somatotrophs, it may act through a cyclase which is
    diVerent from that used by GHRP-2, since GHRP-2 and
    GHRH have an additive eVect on both cAMP accumulation
    and GH secretion when both secretagogues are
    applied at maximal doses (Wu et al. 1994b, 1996).
    In summary, GHRP-2 promotes cAMP accumulation
    in ovine but not in rat somatotrophs via activation of
    adenylyl cyclase. This appears to be the pathway responsible
    for stimulation of GH secretion by GHRP-2.
    GHRP-6 and GHRP-1 do not, however, increase cAMP
    levels in ovine, rat or human somatotrophs but do amplify
    the cAMP response to GHRH in rat pituitary cells. This
    amplification may explain the synergistic action of
    GHRP-6 or GHRP-1 and GHRH on GH secretion in
    this species.
    Protein kinase C (PKC) It has been suggested that the
    action of GHRP-6 and the non-peptidergic analogue
    L-692,429 to stimulate GH release from rat pituitary cells
    is mediated by PKC (Cheng et al. 1991, 1993). The
    synergistic eVect of GHRP-6 and GHRH on cAMP
    accumulation and GH secretion in rat pituitary cells may
    also be mediated by PKC (Cheng et al. 1991). It should be
    noted, however, that the specificity of the inhibitor
    (phloretin) used in the latter study has not been widely
    tested and its eVect on other kinase systems is not defined.
    In particular, over the same dose range (10–200 ΅M),
    phloretin increased the opening probability of Ca2+-
    activated K+ channels (Koh et al. 1994) which can
    hyperpolarize cell membrane potential and prevent the
    stimulation of GHRP-6 on GH secretion. Downregulation
    of PKC with long-term treatment by
    phorbol,12-myristate,13-acetate (PMA; 1 ΅M) partially
    blocked the eVect of GHRP-6 on GH secretion (Cheng
    et al. 1991), suggesting some involvement of PKC. It was
    shown by Akman et al. (1993) however, that GHRP-1
    causes GH release following maximal stimulation of cells
    with PMA. In ovine pituitary cells, GHRP-6 does not
    Commentary 383
    Journal of Endocrinology (1996) 148, 381–386
    cause PKC translocation (Wu et al. 1995). Furthermore,
    down-regulation of PKC with PMA does not block GH
    release in response to GHRP-6 whereas PMA-stimulated
    GH release is totally abolished by the same treatment (Wu
    et al. 1995).
    In contrast, GHRP-2 stimulates PKC translocation
    from cytosol to membrane in ovine somatotrophs in
    primary culture (Wu et al. 1995). PKC inhibitors (calphostin
    C, chelerythrine, staurosporine) and down-regulation
    of PKC by phorbol-12,13-dibutyrate causes partial attenuation.
    It seems likely therefore, that PKC is at least partially
    involved in the action of GHRP-2 (but not GHRP-6) in
    sheep cells. It is interesting to note that GHRH also causes
    PKC translocation in ovine somatotrophs (Wu et al.
    1995). GH secretion in response to GHRH and GHRP-2
    can be accounted for by activation of the cAMP/PKA
    pathway in ovine somatotrophs, and the concomitant
    activation of PKC may play a role in the regulation of GH
    synthesis.
    InsP3 GHRP-6 and GHRP-1 increase [Ca2+]i through
    mobilization of intracellular stores through a mechanism
    that probably involves InsP3 (Bresson-Bepoldin & Dufy-
    Barbe 1994, Herrington & Hille 1994). Indeed, it has been
    reported that GHRP-6 and non-peptidergic GH secretagogues
    increased phosphatidylinositol (PI) turnover via
    an activation of phospholipase C (PLC) in human acromegalic
    tumour cells (Adams et al. 1995, Lei et al. 1995).
    Activation of PLC will produce both InsP3 which leads to
    the release of Ca2+ from intracellular Ca2+ storage sites,
    and diacyglycerol (DAG) which activates PKC. Whether
    PLC (or an increase in PI turnover) is activated in ovine or
    rat somatotrophs by any GHRP is still an open question.
    Investigation of this issue would be useful given the
    diVerence in the response of sheep cells to GHRP-2 and
    other versions of GHRPs.
    Receptor/s for GHRPs
    It seems likely that a novel endogenous receptor exists
    for GHRPs although there is, as yet, no indication of
    its/their identity. It is certainly clear that GHRPs do not
    act through the GHRH receptor. Evidence for this is
    as follows. (1) A GHRH receptor antagonist inhibits
    GHRH-stimulated GH release but not GHRP-6- or
    GHRP-1-stimulated GH release (Thorner et al. 1994, Wu
    et al. 1994a). A putative GHRP receptor antagonist does
    not aVect GH release in response to GHRH (Cheng et al.
    1989, Thorner et al. 1994). (2) Use of a radioreceptor assay
    for GHRH indicates that GHRP-6 does not compete with
    the GHRH-binding sites (Thorner et al. 1994). (3) Functionally,
    there is an additive eVect on GH release when
    both GHRH and the GHRPs are co-administered at a
    maximal dosages (Wu et al. 1994a,b). (4) There is no
    cross-desensitization between GHRH and GHRPs
    whereas homologous desensitization occurs (Wu et al.
    1994a,b). Because GHRPs stimulated GH release in vitro
    in the absence of SRIF, it is unlikely that GHRPs act as
    an inverse agonist on SRIF receptor. All of this evidence
    strongly suggests that there is a novel receptor for
    GHRPs.
    It should, however, be emphasized that the GHRH
    antagonist ([Ac-Tyr1,D-Arg2]-GHRH1–29) did prevent
    GH release in response to GHRP-2 in sheep pituitary cells
    (Wu et al. 1994b) which suggests some relationship
    between the GHRP-2-binding sites and the GHRH
    receptor. Previously, GHRP-6 was reported to interact
    with a novel low-aYnity GHRH-binding site in rat
    anterior pituitary cell membranes (Lau et al. 1991). Meanwhile,
    Sethumadhavan et al. (1991) have identified two
    classes of GHRP-binding sites using [125I]Tye-Ala-
    GHRPs as a ligand. It seems possible therefore that a
    receptor exists that has a low aYnity for GHRH and a high
    aYnity for GHRP-2. Whether there is some homology
    between receptors for GHRH and GHRPs is not known.
    Another possibility that we considered was that GHRPs
    could bind to a site on the GHRH receptor diVerent from
    that employed by GHRH. This was tested using a GC cell
    line with over-expression of the GHRH receptor that was
    kindly provided by Professor S Melmed (C Chen, J
    Zhang, P Farnworth, B Canny, S Petersenn & I J Clarke,
    unpublished observations). In these cells, GHRH increased
    cAMP levels but GHRP-2 or GHRP-6 did not do
    Figure 1 The proposed signalling pathways for GHRPs in
    somatotrophs. The binding of GHRPs to a putative receptor
    activates the PLC and adenylyl cyclase pathways via G-proteins,
    leading to an increase in InsP3 and the activity of PKC and PKA.
    InsP3 then releases Ca2+ from the InsP3-sensitive Ca2+ pool and
    protein kinases phosphorylate ion channels to increase Ca2+
    influx. All of these events would lead to an increase in [Ca2+]i and
    GH secretion. G=GTP-binding proteins; PIP2=phosphatidylinositol
    (4,5)-bisphosphate.
    Commentary 384
    Journal of Endocrinology (1996) 148, 381–386
    so; the GHRH eVect was blocked by a GHRH receptor
    antagonist. This suggests that GHRPs do not act on
    GHRH receptors. Nevertheless, it remains possible that
    there is more than one GHRP receptor, type one
    (GHRP-R1) that is not blocked by the GHRH antagonist
    ([Ac-Tyr1,D-Arg2]-GHRH 1–29) and type two (GHRPR2)
    that may be blocked. There is a possibility that
    GHRP-R1 has a similar binding aYnity for GHRP-6,
    GHRP-1 and GHRP-2 whereas GHRP-R2 would have
    a higher aYnity for GHRP-2 than for GHRP-6 or
    GHRP-1. GHRP-R2 may also have a low aYnity for
    GHRH-binding which would explain the eVects of
    blockade with [Ac-Tyr1,D-Arg2]-GHRH 1–29.
    The GHRP receptors (both GHRP-R1 and -R2) are
    most likely to be coupled to G-proteins based on the
    evidence of activation of adenylyl cyclase by GHRP-2
    (Wu et al. 1996), activation of PKC (Cheng et al. 1991,
    Wu et al. 1995), release of intracellular Ca2+ (Bresson-
    Bepoldin & Dufy-Barbe 1994, Herrington & Hille 1994)
    and an increase in PI turnover by GHRP-6 or nonpeptidergic
    secretagogues (Adams et al. 1995, Lei et al.
    1995). Because both adenylyl cyclase and phospholipase C
    (PLC) are activated by GHRPs, Gs and Gq are most likely
    to be involved in the response.
    Conclusion
    As discussed above, two major signal pathways have been
    identified in somatotrophs employed by diVerent forms of
    GHRP in diVerent animal species. The diVerences in
    signalling systems may reflect two subtypes of receptor for
    GHRPs. Because of cross-talk between diVerent signalling
    systems, Fig. 1 illustrates the signalling pathways employed
    by both possible GHRP receptors with GHRP-R1 mainly
    coupled to PKC pathway and GHRP-R2 mainly to PKA
    pathways.
    The therapeutical use of GHRP is quite obvious but its
    physiological role is still a mystery. Although the roles of
    GHRH and SRIF are well established as endogenous
    regulators of GH, the model of reciprocal secretion that
    was proposed for the rat (Tannenbaum & Ling 1984)
    cannot fully explain the pattern of GH secretion pattern in
    sheep and human. Studies on the mechanism of the action
    of synthetic GHRPs has revealed a novel regulatory system
    for the manipulation of GH secretion. The potency of
    GHRPs to release GH is higher in human and sheep than
    in the rat. DiVerences in signal transduction pathways have
    also been demonstrated between sheep and rat. A heterogeneity
    of function is therefore suggested among diVerent
    species. It seems unlikely that nature would create a
    redundant system or a system that abets therapeutical use,
    so we must conclude that an endogenous ligand exists for
    GHRP receptors. This endogenous ligand for GHRP
    receptors may play an important role in the regulation of
    GH secretion in concert with GHRH and SRIF. The
    identification of endogenous GHRPs ligands and receptors
    provides a challenge for the future.
  

  
 

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