miquelangeles

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For the last ~6 months I've been experimenting with pregnenolone in different dosages, frequencies, schedule and routes of administration.

There are already numerous threads about it's effects (they are very different depending on all those factors), but I want to mention two things that I haven't seen discussed here:

1. Some metabolites like DHEA and DHEA-S have a half life of 12-24 hours, which means they might keep accumulating for 5 days until reaching steady state concentration. It may explain why sometimes the effects are not consistent every day. Dosing every other day or once a week has been mentioned here, and it might help in this case.

2. I noticed that I feel best when taking it on an empty stomach, usually first thing in the morning. Taken at night, it is hit and miss. Sometimes it would cause insomnia, other nights I would sleep great and wake up feeling amazing. Taking it too close to dinner might also be a factor. But I noticed that taking it at night 1-2 hours before going to bed feels very different than taking it right at bedtime when lying down. This made me think that perhaps the preg reacts with the hydrochoric acid, since it stays for much longer in the stomach when in a lying position. So I did a quick search on "pregnenolone" + "hydrochloric acid" and this (rat) study came up instead.

If this study translates to humans, it looks like there is another route of pregnenolone or progesterone administration for androgenic effects, and that is trans-stomachal. The rate of conversion depends on the number of parietal cells exposed, so it's best to swallow the powder directly and chase it with a little bit of water just enough to distribute the preg to a larger surface of the stomach and not send it too far down in the duodenum or small intestine. Maybe this is also how Ray was taking it, since he was using bulk powder and I doubt he was bothering to encapsulate it.


Androgen biosynthesis in the stomach: expression of cytochrome P450 17 alpha-hydroxylase/17,20-lyase messenger ribonucleic acid and protein, and metabolism of pregnenolone and progesterone by parietal cells of the rat gastric mucosa

Dehydroepiandrosterone (DHEA) and its conjugates persist in the rat brain, for up to 1 month after ablation of both adrenals and gonads. Since DHEA synthesis in brain from pregnenolone (PREG) was excluded, we have considered other tissular sources including the digestive tract. In situ hybridization with specific oligonucleotide probes showed that the parietal cells of the gastric mucosa, contrary to other cell types, strongly expressed P450(17) alpha messenger RNA. Expression of the enzyme in the parietal cells was confirmed by immunocytochemistry with specific antibodies. An intense reaction was observed in the stomach of adult males and of cyclic or pregnant females. Access to food did not influence the intensity of immunostaining. It appeared at postnatal days 16-21, then the number of positive cells increased rapidly and leveled off at adult age. Parietal cells were released by pronase digestion of everted stomachs from adult male and female rats and were purified by density gradient centrifugation on Nycodenz. 5 x 10(4) to 1.6 x 10(6) cells were incubated with either 1 microM 14C-PREG or 14C-progesterone (14C-PROG) at 37 C under 95% O2-5% CO2, for 10-180 min. PREG was converted to 17-OH PREG and to androstenediol, whereas PROG was converted to 17-OH PROG and to testosterone. Only minute amounts of either DHEA or androstenedione, respectively, were detected at any incubation time, indicating their fast conversion to the corresponding 17 beta-hydroxysteroids. 3H-25-OH cholesterol was not metabolized to 3H-PREG, and 14C-PREG was not converted to 14C-PROG, in accordance with negative immunocytochemical results with antibodies to cytochrome P450scc and 3 beta-hydroxysteroid dehydrogenase delta 5-->4-isomerase (3 beta-HSD). In conclusion, the parietal cells, which are known as the source of gastric acid secretion, can synthesize testosterone from PROG and androstenediol from PREG. The physiological relevance of such conversions remains to be established.

...

This report was initiated by the search of the origin of brain DHEA (as the sulfate and fatty acid esters) because it persists in rats deprived of steroidogenic glands. DHEA cannot be formed in the rat brain from PREG because the P450 17alpha enzyme is lacking.
...
The P450 17alpha enzyme is expressed in the rat stomach at the mRNA, protein, and activity level. Both in situ hybridization and immunocytochemistry have shown that P450 17alpha is expressed
solely in the parietal cells of the gastric mucosa; no labeling occurred either in the muscular layer or in the zymogenic chief cells or in mucous cells of the glandular mucosa.
...
The P450 17alpha protein was detected in the parietal cells of both male and female rats with no obvious differences in signal intensity. It was not expressed in the parietal cells of the rat until the third week of life, which is the weaning period. Whether the expression of l’4501za is under dietary influence, or merely corresponds to a functional maturation of the parietal cells, remains to be determined.
It is particularly intriguing that the gene
...
DHEA formed from PREG was shown in the present to be readily converted into androstenediol. The reverse conversion from androstenediol to DHEA might occur in brain cells.
...

The capacity of the stomach to produce testosterone from PROG is intriguing and merits further studies in search for an eventual endocrine function, at least in absence of testicular secretion. Another possibility would be that parietal cells or other type(s) of gastric mucosal cells contain androgen receptors and that testosterone synthesized in situ from circulating PROG would exert an autocrine or paracrine function.
...

time-course-C14-PROG.jpg
@Makrosky @Mauritio
 

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miquelangeles

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Extraglandular hormonal steroidogenesis in aged rats

Abstract​

We have examined the metabolism in vitro of [4-14C]pregnenolone by the following organs of 2.4-year-old rats: submandibular gland, stomach, duodenum, liver, lung, heart, spleen, kidney, skin, prostate, testis and adrenal. All tissues converted pregnenolone to progesterone, the highest yields being observed with adrenal, testis and skin. Androgen formation was intense in the testis and absent in the adrenal. Moreover, 17α-hydroxylation of pregnenolone occurred moderately in kidney, skin and submandibular gland and markedly in duodenum and stomach, which also produced high amounts of dehydroepiandrosterone and/or 5-androstene-3β,17β-diol. Extratesticular synthesis of androstenedione and testosterone was very low. A significant formation of 20α-dihydropregnenolone was observed in all tissues but stomach, duodenum and steroidogenic endocrines. Corticosteroids were not synthesized extraadrenally, except a small amount of 11-deoxycorticosterone in the testis. These results indicate that key steroid-biosynthetic enzymes, such as 3β-hydroxysteroid dehydrogenase/Δ5′Δ4 isomerase, 17β- and 20α-hydroxysteroid dehydrogenases and steroid 17α-monooxygenase/17,20-lyase are also expressed extraglandularly in the rat.
 

miquelangeles

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Occurrence of cytochrome P450c17 mRNA and dehydroepiandrosterone biosynthesis in the rat gastrointestinal tract

Abstract​

We have investigated 17α-hydroxylase and C17,20-lyase activities and the presence of cytochrome P450c17 mRNA in the esophagus, stomach, duodenum, and colon of adult rats of both sexes. All tissues converted [4-14C]pregnenolone mainly to dehydroepiandrosterone (DHEA) through the 5-ene-3β-hydroxysteroid route as opposed to the 4-ene-3-ketosteroid pathway in a control testicular incubate. Synthesis of dehydroepiandrosterone was particularly high in the duodenum and was found to be lower in the stomach, colon and esophagus, in decreasing order. 20α-Hydroxypregnenolone and progesterone were also formed primarily by the esophagus and colon, respectively. P450c17 mRNA was demonstrated by ribonuclease protection assay in the stomach and duodenum, but not in esophagus and colon. However, a 335 bp-long cDNA fragment, whose sequence corresponded to that of rat P450c17 cDNA, was amplified by reverse transcription (RT) and polymerase chain reaction (PCR) from the poly(A)+ RNAs of all four tissues. This result was further confirmed by Southern blotting using a 794-bp testicular probe. The complete sequence of P450c17 cDNA in the stomach and duodenum was identical to that reported for rat testis P450c17 cDNA. No amplification and no positive signal in Southern blotting were observed with the total RNAs from adult male adrenal and spleen, which were taken as negative controls since they had been previously found unable to form androgens from pregnenolone. Although the levels of transcription in gonads, duodenum and stomach were found to be equivalent, as indicated by the RNase protection assay and semiquantitative RT-PCR assay, P450c17 enzyme activity was much higher in the testis, pointing at a possible dissimilarity in the respective rates of mRNA translation. Thus, P450c17 is differentially expressed in the rat gastrointestinal tract, where it leads to the synthesis of the sex steroid precursor DHEA, especially in the duodenum and stomach.
 

Makrosky

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Very interesting miquelangeles, thanks!

I think though the differences you experience are more like related to the cyrcadian rhythms? Like different times of the day/night favor certain conversion routes or others?

Also, even if it is not about that, let's take the pregnenolone example assuming it lowers cortisol. Cortisol follows a pattern during the day, if you lower it too much or leave it unoposed at different times of the day, it will have different subjective effects.

Btw could you try intranasal DHEA?

@haidut
 

miquelangeles

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Very interesting miquelangeles, thanks!

I think though the differences you experience are more like related to the cyrcadian rhythms? Like different times of the day/night favor certain conversion routes or others?

Also, even if it is not about that, let's take the pregnenolone example assuming it lowers cortisol. Cortisol follows a pattern during the day, if you lower it too much or leave it unoposed at different times of the day, it will have different subjective effects.

Btw could you try intranasal DHEA?

@haidut
Yes, circadian rhythms should definitely be taken into account. That's why I tried to take it at night, since the majority of the daily T release occurs during sleep. Plus the supposed cortisol lowering effect.
I believe the favoring of certain routes depending on time of day probably applies to circulating plasma levels mostly. Because it seems that as soon as it gets in contact with the mouth, esophagus, stomach, liver or skin it readily gets converted to other metabolites depending on which enzymes are expressed in the respective tissue. And this seems to happen when incubated with isolated cells too, irrespective of time of the day.
So like you said, it is more about aligning the timing/dose/route with the existing circadian rhythms.
I tried intranasal DHEA and didn't like it, felt just like sublingual which slows down my thinking and makes me feel weird. I feel best with encapsulated DHEA bypassing the stomach so I probably get benefit from the DHEA-S. Intanasal preg on the other hand, feels great. I haven't done it often because of the rice flour filler in capsules, but I'm planning to get some from Cayman Chemical Europe, their price is 100 euro for 10 grams.
 

Makrosky

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Yes, circadian rhythms should definitely be taken into account. That's why I tried to take it at night, since the majority of the daily T release occurs during sleep. Plus the supposed cortisol lowering effect.
I believe the favoring of certain routes depending on time of day probably applies to circulating plasma levels mostly. Because it seems that as soon as it gets in contact with the mouth, esophagus, stomach, liver or skin it readily gets converted to other metabolites depending on which enzymes are expressed in the respective tissue. And this seems to happen when incubated with isolated cells too, irrespective of time of the day.
So like you said, it is more about aligning the timing/dose/route with the existing circadian rhythms.
I tried intranasal DHEA and didn't like it, felt just like sublingual which slows down my thinking and makes me feel weird. I feel best with encapsulated DHEA bypassing the stomach so I probably get benefit from the DHEA-S. Intanasal preg on the other hand, feels great. I haven't done it often because of the rice flour filler in capsules, but I'm planning to get some from Cayman Chemical Europe, their price is 100 euro for 10 grams.
Interesting. Does Cayman sell retail style to private individuals? Looks like a cheaper version of Sigma Aldrich. 100 euro for 10 grams is kind of okish if the purity is really USP. Will last a long time if used only intranasal.

Edit: Not sure how those 10 grams would drgrade over time.
 

miquelangeles

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Interesting. Does Cayman sell retail style to private individuals? Looks like a cheaper version of Sigma Aldrich. 100 euro for 10 grams is kind of okish if the purity is really USP. Will last a long time if used only intranasal.

Edit: Not sure how those 10 grams would drgrade over time.
They only sell to companies, but I’m ordering through a friend who has a medical supply business. There is also FSA Chemicals in Belgium who sell 5g for 45 euro, but their minimum order is 250euro.
 

Mauritio

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Interesting.

Looks like like oral progesterone seems especially good for increasing T.

I get good results from tiny amounts of Prog and preg (less than 1mg) taken orally.
And I always thought they had a more liver boosting / androgenic effect taken orally . I ascribed it to their anti-endotoxin effect,but your theory makes a lot of sense too!
 

miquelangeles

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Interesting.

Looks like like oral progesterone seems especially good for increasing T.

I get good results from tiny amounts of Prog and preg (less than 1mg) taken orally.
And I always thought they had a more liver boosting / androgenic effect taken orally . I ascribed it to their anti-endotoxin effect,but your theory makes a lot of sense too!
But the tiny amount you take orally can actually be considered sublingual? It doesn’t get processed during the first pass of the liver. I sometimes put a small amount of prog in a capsule and swallow it, and it feels better than taking it sublingually/through the gums. The liver preferentially converts it to allo and it’s not as sedating as taking the same amount sublingually. And I tried prog intranasally and it didn’t feel sedating. I wonder what enzymes are expressed sublingually. Similarly, a study found sublingual androstenediol raises serum T, but swallowing does not.
 

Makrosky

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Fascinating. It never occurred to me that some of the sublingual route effects (not only for hormones but anything) which are different from oral, could be in part because of different enzymes expressed on the tissues of that area.
 

miquelangeles

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Fascinating. It never occurred to me that some of the sublingual route effects (not only for hormones but anything) which are different from oral, could be in part because of different enzymes expressed on the tissues of that area.
Well it's in the same way as you pointed it out first in the "skin as steroidogenic organ" thread. This doesn't mean there isn't direct absorption through oral mucosa as well, or other mechanisms involved, but there is evidence at least for esophagus and stomach that they metabolize pregnenolone. So it's likely the oral mucosa does too.
 

haidut

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For the last ~6 months I've been experimenting with pregnenolone in different dosages, frequencies, schedule and routes of administration.

There are already numerous threads about it's effects (they are very different depending on all those factors), but I want to mention two things that I haven't seen discussed here:

1. Some metabolites like DHEA and DHEA-S have a half life of 12-24 hours, which means they might keep accumulating for 5 days until reaching steady state concentration. It may explain why sometimes the effects are not consistent every day. Dosing every other day or once a week has been mentioned here, and it might help in this case.

2. I noticed that I feel best when taking it on an empty stomach, usually first thing in the morning. Taken at night, it is hit and miss. Sometimes it would cause insomnia, other nights I would sleep great and wake up feeling amazing. Taking it too close to dinner might also be a factor. But I noticed that taking it at night 1-2 hours before going to bed feels very different than taking it right at bedtime when lying down. This made me think that perhaps the preg reacts with the hydrochoric acid, since it stays for much longer in the stomach when in a lying position. So I did a quick search on "pregnenolone" + "hydrochloric acid" and this (rat) study came up instead.

If this study translates to humans, it looks like there is another route of pregnenolone or progesterone administration for androgenic effects, and that is trans-stomachal. The rate of conversion depends on the number of parietal cells exposed, so it's best to swallow the powder directly and chase it with a little bit of water just enough to distribute the preg to a larger surface of the stomach and not send it too far down in the duodenum or small intestine. Maybe this is also how Ray was taking it, since he was using bulk powder and I doubt he was bothering to encapsulate it.



@Makrosky @Mauritio

Yep, the GI tract is a steroidogenic organ (actually all cells are since they all express at least the side-chain cleavage enzyme and aromatase). I remember seeing a few studies saying the GI tract synthesizes primarily androgens and cortisol. The latter is probably done to keep inflammation low from the constant endotoxin assault the GI tract experiences. Aspirin metabolizes into salicylic acid, which blocks the final step of cortisol formation so taking pregnenolone with aspirin may work even better. Adding some niacinamide to raise NAD levels (a cofactor for many of the steroidogenic enzymes) may help even more and amplify the androgenic pathway/effects while inhibiting the estrogenic ones.

@Makrosky
 
Last edited:

haidut

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Thanks @haidut I mentioned you as I thought you would find the studies miquelangeles posted interesting.

Thanks, I was not complaining in any way that you tagged me:):
Btw, I added one more link (last one) to my post in support of the pregnenolone + niacinamide suggestion.
 

miquelangeles

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Yep, the GI tract is a steroidogenic organ (actually all cells are since they all express at least the side-chain cleavage enzyme and aromatase). I remember seeing a few studies saying the GI tract synthesizes primarily androgens and cortisol. The latter is probably done to keep inflammation low from the constant endotoxin assault the GI tract experiences. Aspirin metabolizes into salicylic acid, which blocks the final step of cortisol formation so taking pregnenolone with aspirin may work even better. Adding some niacinamide to raise NAD levels (a cofactor for many of the steroidogenic enzymes) may help even more and amplify the androgenic pathway/effects while inhibiting the estrogenic ones.

@Makrosky
Thank you for replying. I am often taking the pregnenolone with niacinamide, but I never tried it with aspirin because I usually don't feel much from aspirin alone.
But I've seen the combination mentioned here several times and I read that it was actually Ray who recommended it to someone over email? What dose of aspirin do you think is good?
 

haidut

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Thank you for replying. I am often taking the pregnenolone with niacinamide, but I never tried it with aspirin because I usually don't feel much from aspirin alone.
But I've seen the combination mentioned here several times and I read that it was actually Ray who recommended it to someone over email? What dose of aspirin do you think is good?

He recommended aspirin and pregnenolone. The niacinamide as another addition is my guess based on the studies above. As far as aspirin, I think he said 2-3 tablets in cases of severe stress/inflammation may be needed. There is a human study showing 800mg twice a day, by itself, raised T levels and lowered cortisol in human males, which seems to corroborate the other study showing effects on the 11b-HSD1 enzyme. When combined with pregnenolone, it is plausible that even 1 tablet (325mg) twice daily would have similar effects as the higher dose in the study below.
 

Mauritio

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But the tiny amount you take orally can actually be considered sublingual? It doesn’t get processed during the first pass of the liver. I sometimes put a small amount of prog in a capsule and swallow it, and it feels better than taking it sublingually/through the gums. The liver preferentially converts it to allo and it’s not as sedating as taking the same amount sublingually. And I tried prog intranasally and it didn’t feel sedating. I wonder what enzymes are expressed sublingually. Similarly, a study found sublingual androstenediol raises serum T, but swallowing does not.
Yes that's a good point, its probably more sublingual than oral, especially since I use a powder. But I dont find it sedating at all. These tiny doses are more stimulating for me than larger ones.
 
Progesterone - Pregnenolone - DHEA- Salicylic Acid - Urea - Cholesterol Powder - Artemisia

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