TRT Effect on 5 Alpha Reductase and Upstream Hormones- Cause of Low Mood and Anxiety?

[Gianluca]

Pharmacology of pregnenolone and progesterone:

"Oral administration of 50 or 100 mg pregnenolone has been found to have minimal or negligible effect on urinary levels of testosterone and testosterone metabolites, including of androsterone, etiocholanolone, 5-androstanediol, androstadienol, and androstenol (and/or their conjugates), and this suggests that only a small amount of pregnenolone is converted into testosterone.[17][18] This is in accordance with findings on the conversion of DHEA into testosterone, in which only 1.5% of an oral dose of DHEA was found to be converted into testosterone.[17] In contrast to the androstanes, 50 or 100 mg oral pregnenolone has been found to significantly and in fact "strongly" increase urinary levels of the progesterone metabolites pregnanediol and pregnanolone (and/or their conjugates), whereas pregnanetriol was unaffected.[17][18] Unlike the case of oral administration, transdermal administration of 30 mg/day pregnenolone cream has not been found to affect urinary levels of metabolites of any other steroids, including of progesterone.[18]"

Pregnenolone - Wikipedia

Pregnenolone and Progesterone for Men: Pharmacokinetics and Studies

"Pregnenolone can be administered orally, subcutaneously, intravenously, intranasally, and topically/transdermally. Oral pregnenolone has high metabolism and low bioavailability.28 It is lipophilic and readily crosses the blood brain barrier. There is very limited data on the pharmacokinetics...

www.excelmale.com

Great report! This confirm my personal anecdotal of Pregnenolone cream alone not increasing Progesterone level.

 

[Fortunate]

Thank you.

I have been reluctant to supplement upstream hormones since I have been using hCG plus TRT for years. But I am now in the mood to actually experiment to see if taking pregnenolone plus or minus progesterone can improve mood and sleep without any adverse effects. You have been leading the way on this subject, so I am learning from your experiences.

I have a strong feeling that the TRT field has become complacent (researchers, physicians, etc) with men that do not respond well to TRT (those with increased anxiety on TRT in particular). It would be nice to see if we can collectively do our own open label pilot on these upstream neuro-steroids that are dramatically decreased by TRT and anabolic steroids, specially as hCG access becomes more problematic with FDA restrictions on compounding pharmacies.

Completely agree with looking at these other hormones. I will be watching your guys' response closely and will try to pitch in to the collective knowledge.

My goals:

• Stop hCG if possible (less injections)
• Get TRT benefits without headaches (literal headaches)
• Get sustained TRT benefit without eventual neuropsychological issues like anxiety

I bought some Pregnenolone, but will likely wait until I see more feedback from you guys.

 

[M.J]

Major issue in trt is libido i hope this can help, I have lowered my hcg and for some improvement in libido. Not consistent but still experimenting with it.

 

[MarcoFL]

Thank you.

I have been reluctant to supplement upstream hormones since I have been using hCG plus TRT for years. But I am now in the mood to actually experiment to see if taking pregnenolone plus or minus progesterone can improve mood and sleep without any adverse effects. You have been leading the way on this subject, so I am learning from your experiences.

I have a strong feeling that the TRT field has become complacent (researchers, physicians, etc) with men that do not respond well to TRT (those with increased anxiety on TRT in particular). It would be nice to see if we can collectively do our own open label pilot on these upstream neuro-steroids that are dramatically decreased by TRT and anabolic steroids, specially as hCG access becomes more problematic with FDA restrictions on compounding pharmacies.

One thing I would highly recommend you guys that are interested in your sleep performance is the Oura ring! I recently bought one for the sole purpose of my sleep monitoring. I really helps you to see as you adjust or add things how it impacts your stages of sleep. I have learned a lot with it! I have also just started adding pregnenelone 5mg orally at night to see how that impacts my deep sleep. When I have tested my levels in the past they are very low.

 

[Nelson Vergel]

@Mark Saur you are the third person that mentions the Oura ring. Is there a subscription fee to access data?

 

[MarcoFL]

@Mark Saur you are the third person that mentions the Oura ring. Is there a subscription fee to access data?

I do not recall for sure but was not very much if there was a charge. It is very easy to use and I do not wear it during the day only for sleep. I have learned a lot from it the last few months! I cannot recommend it enough if you are obsessed like I am and want to perfect sleep as much as possible.

 

[DixieWrecked]

I'm right there with you guys on trying to figure this out. Do you ever think the elevated estrogen is causing issues? I bit the bullet about a week ago, a little more, and started arimidex. I feel a whole lot better. Way less anxiety. I notice my conversations are calmer. My penis is functioning way better.

I know the general consensus on ais is to avoid them but I am willing to accept it if thats what it takes.

 

[Nelson Vergel]

I know the general consensus on ais is to avoid them but I am willing to accept it if thats what it takes.

I think if anything works for anyone, it does not matter really what the data says. We only know one thing: sustained low estradiol (under 12 pg/mL in one well done study) can eventually cause loss of bone density, increased fat mass, and loss of libido. We do not know what happens with high estradiol, although we know that a low testosterone-to-estradiol ratio may impair fertility and increase the chance for gynecomastia (specially in the presence of high IGF-1).

 

[Anonymon]

Excellent! I’m deciding to stick with the cream in the am since it helps me with energy prior my work out better than the capsule, and I will take the 50mg capsule at night since it quickly converts into Progesterone, I think it makes more sense this way. I really love the cream, I hope it works well for you as well.

Empower had to make a special order for my 5mg Progesterone, because in fact they don’t carry this strength

I didn’t notice any testicular volume improvement.

You certainly know and thought all of us how to use and manage hormones for health, but I still feel obligated to say this: if you change to cream/cap protocol, I would wait a week or two before introducing/trying Progesterone cap

Since Post Finasteride Syndrome seemingly lead me to all this, I’m going to try your protocol to the extent I can, though I’ll be limited to what’s on Amazon starting off. The cream gives you energy? I’ve tried progesterone cream, not pregnenolone cream, off and on in the evening and it always calms me and makes me more tired. Pregnenolone cream is the opposite then I take it?

I’ve also found I do better when I have a high amount of saturated fat in my diet—particularly MCT oil as of late—though I have more of a tendency to put on unwanted weight when doing that.

 

[Charliebizz]

I should be getting results from a full work up this week and can share my results also. On test e only. My ferritin has dropped from 157 natural to 50. Will see if it has dropped further. I also experience headaches and low mood on trt.

Got my labs back my ferritin actually went back up to 70 with really no change in protocol other then a minor dose decrease from 90 to 84mg a week.

My one issue seems to be with free t. I have low shbg. Ive tested 8 hours after injection and 28 hours after. I inject m-w-f. 28-30mg an injection. 8 hours after injection my tt was 900 and ft 200 (35-155) and 28nhoura after injection my tt drops to 500-550. But ft only droped to 160. I do notice the headaches and mood issues more on injection day so maybe the high free t is the cause. I wonder how low I can drop my dose to still get the benefits ( libido and recovery) without the headaches. Seems I need to forget about total t and focus on free t only.

 

[Gianluca]

Since Post Finasteride Syndrome seemingly lead me to all this, I’m going to try your protocol to the extent I can, though I’ll be limited to what’s on Amazon starting off. The cream gives you energy? I’ve tried progesterone cream, not pregnenolone cream, off and on in the evening and it always calms me and makes me more tired. Pregnenolone cream is the opposite then I take it?

I’ve also found I do better when I have a high amount of saturated fat in my diet—particularly MCT oil as of late—though I have more of a tendency to put on unwanted weight when doing that.

Saturated Fats are healthy, even because they help to raise Cholesterol, as we understand the steroid hormones pathways. The cream gives me more mental and physical energy, I feel better on my work outs when I use it, but it is also calming at same time. The energy it gives me it is not like caffeine type, I take Pregn cream some time at night as well, and it helped to fall a sleep faster and deeper. Some people can't really take Pregnenolone during the day as they get drowsy, try see how it works for you

 

[Gianluca]

I'm right there with you guys on trying to figure this out. Do you ever think the elevated estrogen is causing issues? I bit the bullet about a week ago, a little more, and started arimidex. I feel a whole lot better. Way less anxiety. I notice my conversations are calmer. My penis is functioning way better.

I know the general consensus on ais is to avoid them but I am willing to accept it if thats what it takes.

have you checked your Progesterone level before taking an AI?

 

[Fortunate]

for Pregnenolone I use 50mg cap plus 50mg cream (empower) I have tried the cream at night for the past two days, it worked great for sleep. Progesterone 5mg cap generally once per day at night or as needed

@Gianluca, thanks for the information. I have watched most of Dave Lee's two presentations on Pregnenolone and DHEA. I have to admit I am getting caught in analysis paralysis. I am not sure what to make of the information out there and how to proceed. For example, I have noticed very definite fatigue and sleepiness when taking DHEA. I have also noticed it darkness my mood. Yet, he purports DHEA does the exact opposite: take it in the morning, as it gives you energy and it is a strong antidepressant. He does provide the disclaimer that both DHEA and Pregnenolone can have opposite effects in different individuals, but I have a hard time making sense of it. He also says Pregnenolone has a sedative affect, but if you do a little mini experiment on Amazon and read user reviews of OTC Pregnenolone, many say the opposite: Be careful, it amps you up!

I have a particular interest in this because I have previously experienced "The Wall" - after doing very well on Xyosted for months, I started feeling anxious, overstimulated, overall bad. I am now back on Xyosted and doing fine, but would like something to combat that feeling if it happens again. This time, I am trying it without hCG (I am still undecided on whether or not hCG causes me problems). However, if I hit the Wall again, it would be nice to know there are options to address it. I have been around long enough to know there are typically no silver bullets out there for any one problem. But, I became intrigued when I saw Pregnenolone being discussed specifically as a way to address the anxiety issues one gets after a while on TRT (The Wall).

In any case, can you give your thoughts time of day? Do you use DHEA in the morning or evening? Have you noticed any sedation? I see you are taking Pregnenolone in the AM as both topical and oral from Empower. Have you ever tried an OTC version?

Thanks again for your thoughts and would love to hear others' experience.

 

[Willyt]

@Gianluca, thanks for the information. I have watched most of Dave Lee's two presentations on Pregnenolone and DHEA. I have to admit I am getting caught in analysis paralysis. I am not sure what to make of the information out there and how to proceed. For example, I have noticed very definite fatigue and sleepiness when taking DHEA. I have also noticed it darkness my mood. Yet, he purports DHEA does the exact opposite: take it in the morning, as it gives you energy and it is a strong antidepressant. He does provide the disclaimer that both DHEA and Pregnenolone can have opposite effects in different individuals, but I have a hard time making sense of it. He also says Pregnenolone has a sedative affect, but if you do a little mini experiment on Amazon and read user reviews of OTC Pregnenolone, many say the opposite: Be careful, it amps you up!

I have a particular interest in this because I have previously experienced "The Wall" - after doing very well on Xyosted for months, I started feeling anxious, overstimulated, overall bad. I am now back on Xyosted and doing fine, but would like something to combat that feeling if it happens again. This time, I am trying it without hCG (I am still undecided on whether or not hCG causes me problems). However, if I hit the Wall again, it would be nice to know there are options to address it. I have been around long enough to know there are typically no silver bullets out there for any one problem. But, I became intrigued when I saw Pregnenolone being discussed specifically as a way to address the anxiety issues one gets after a while on TRT (The Wall).

In any case, can you give your thoughts time of day? Do you use DHEA in the morning or evening? Have you noticed any sedation? I see you are taking Pregnenolone in the AM as both topical and oral from Empower. Have you ever tried an OTC version?

Thanks again for your thoughts and would love to hear others' experience.

Why not just start at lowest dose of 5mg and work your way up depending on how you feel?
I am one of those who can only take DHEA in the morning because of how amped I get even with low doses

 

[madman]

try stopping HCG, and focus to supplement with Pregnenolone first then DHEA, there is no need for HCG to increase Pregnenolone levels. To my understanding ACTH also activate some enzymatic process that converts Pregnenolone into downstream metabolites, but it is possible that LH is needed also, paraphs ACTH activates some, and LH activates some other,

maybe someone who has a better understanding of this can explain, but if Mark Gordon doesn't know yet how to reactivate fully 5AR1, I doubt there is a easy answer to it

Really?

hCG mimics LH which stimulates Leydig cells leading to an increase in both ITT (intratesticular testosterone) and circulating testosterone levels.

LH is critical as it stimulates cAMP production and cholesterol transport to and metabolism within mitochondria.



post#37

Playing with HCG gave me libido

I am thinking more like maybe 500 hcg and 250 is too much and increases something very high where you don’t get this libido maybe low than 250 is a key ? I am checking this now I will see how long it will last M.J., what you're finding is also being discussed on this thread as well...

www.excelmale.com

My reply:

The main purpose of adding hCG to trt is to preserve/maintain fertility and prevent/minimize testicular atrophy.

The use of exogenous testosterone results in the suppression of ITT (intra-testicular testosterone) which is critical for sperm production.

The main goal when using hCG is to restore physiological ITT levels and in order to achieve such a minimum effective dose would be needed (125-500IU) and 250-500IU would seem to be the sweet spot.

Anything less will have a minimal impact on increasing ITT!

Other than one experiencing possible side effects from such doses (250-500IU) using anything <125IU will have a minimal impact on increasing ITT.

My reply from a previous thread where the poster asked if hCG was needed.


Depends on the individual.....Is hCG needed?

*To preserve/maintain fertility then yes.

*To prevent/minimize testicular atrophy then yes.

*To enhance mood/libido than it is not a given as some may experience such effects whereas others may feel worse-off.

*To maintain upstream hormones and possibly prevent long-term consequences for health/well-being.....you be the judge!

Effect of TRT vs hCG/FSH on upstream hormone pathways.

The role of gonadotropins in testicular and adrenal androgen biosynthesis pathways -insights from males with congenital hypogonadotropic hypogonadism on hCG/rFSH and on testosterone replacement SUMMARY Objective: To delineate the role of gonadotropins in male androgen biosynthesis pathways...

www.excelmale.com

*take-home point

A replacement regimen with combined hCG/rFSH mimics physiologic steroid hormone profiles better than a substitution with exogenous testosterone. The documented differences in steroid profiles on testosterone replacement in hypogonadal males with absent or severely reduced endogenous LH and FSH secretion may have long-term consequences for health and wellbeing. Specifically, body composition, bone health, glucose, and lipid metabolism, salt and water balance, cognition, mood, sleep, and sexual function could be affected. The steroidogenic differences could also be relevant for gonadotropin-suppressive treatments with long-acting testosterone preparations in males with primary hypogonadism. To what extent this hypothesis is true, should be addressed in future clinical studies.








Testosterone synthesis in Leydig cells

Steroidogenesis is a multi-step process that converts cholesterol into final steroid hormone products. As reviewed in Aghazadeh and coworkers (Aghazadeh et al. 2015), steroidogenesis consists of cholesterol mobilization from lipid droplets and/or the plasma membrane, cholesterol transport into mitochondria, pregnenolone formation in the mitochondria, and subsequent conversion of pregnenolone into the final steroid products by enzymes of the smooth endoplasmic reticulum (Fig. 1). In the adult testis, Leydig cell TS production depends upon the pulsatile secretion of LH by the pituitary gland into the peripheral circulation. LH plays two essential roles in Leydig cell steroidogenesis: (1) maintenance of optimal levels of steroidogenic enzymes (trophic regulation) and (2) mobilization and transport of cholesterol into the inner mitochondrial membrane (acute regulation).

Leydig cell steroidogenesis.

LH binds its receptor on the Leydig cell membrane. LH receptor/G protein coupling results in increased cAMP and arachidonic acid (AA) production. cAMP stimulates the mobilization and transport of cholesterol to and into the mitochondria in part by activating PKA and MAPK signaling. At the same time, AA can be converted into prostaglandin by Cox2 to negatively regulate the transport of cholesterol across the mitochondrial membranes. At the inner mitochondrial membrane, cholesterol is converted to pregnenolone by CYP11A1, and pregnenolone is converted into testosterone by enzymes in the smooth endoplasmic reticulum (HSD3b, CYP17A1, and HSD17b). Aging and environmental factors may impact steroidogenesis by affecting the intracellular redox balance in part through increased ROS production. This has significant effects on the cAMP formation and/or cholesterol transport into the mitochondria, and thus on steroid formation. Steroidogenesis also might be affected by ROS-induced increases in Cox2 production and redox-sensitive MAPK activation.

Both the trophic and acute effects of LH are mediated by signaling pathways that begin with cAMP production (Fig. 1). LH binds to and activates G protein-coupled receptors, resulting in the activation of adenylyl cyclase, increased intracellular cAMP formation, and cAMP-dependent phosphorylation of proteins through protein kinase A (PKA). The acute stimulation of Leydig cells by LH results in cholesterol transfer into the mitochondria in part through the actions of steroidogenic acute regulatory protein (STAR), translocator protein (18 kDa; TSPO), and other proteins of the transduce some (Midzak et al. 2011). Cholesterol transport into the mitochondria, the rate-limiting step in steroid biosynthesis, is followed by the conversion of cholesterol to pregnenolone by the C27 cholesterol side-chain cleavage cytochrome P450 enzyme (CYP11A1) located on the matrix side of the inner mitochondrial membrane. Pregnenolone then is metabolized into TS by 3β-hydroxysteroid dehydrogenase (3b-HSD; HSD3B), 17α-hydroxylase/17,20 lyase (CYP17A1) and type 3 17β-hydroxysteroid dehydrogenase (17b-HSD3, HSD17B) in the smooth endoplasmic reticulum (Payne & Hales 2004, Aghazadeh et al. 2015, Beattie et al. 2015).

 

[madman]

Pharmacological Regulation of the Cholesterol Transport Machinery in Steroidogenic Cells of the Testis (2015)


Abstract

Reduced serum testosterone (T), or hypogonadism, is estimated to affect about 5 million American men, including both aging and young men. Low serum T has been linked to mood changes, worsening cognition, fatigue, depression, decreased lean body mass and bone mineral density, increased visceral fat, metabolic syndrome, decreased libido, and sexual dysfunction. Administering exogenous T, known as T-replacement therapy (TRT), reverses many of the symptoms of low T levels. However, this treatment can result in luteinizing hormone suppression which, in turn, can lead to reduced sperm numbers and infertility, making TRT inappropriate for men who wish to father children. Additionally, TRT may result in supraphysiologic T levels, skin irritation, and T transfer to others upon contact; and there may be an increased risk of prostate cancer and cardiovascular disease, particularly in aging men. Therefore, the development of alternative therapies for treating hypogonadism would be highly desirable. To do so requires a greater understanding of the series of steps leading to T formation and how they are regulated, and the identification of key steps that are amenable to pharmacological modulation so as to induce T production. We review herein our current understanding of mechanisms underlying the pharmacological induction of T formation in hypogonadal testis.




Introduction

The administration of exogenous T by any means can suppress LH and thus result in reduced Leydig cell T formation and suppression of spermatogenesis. Indeed, contraception in men can be achieved by administering LH-suppressive T. Thus, the exogenous administration of T to ameliorate hypogonadism is inappropriate for men wishing to father children (Carruthers, 2009; Huhtaniemi & Forti, 2011; Perheentupa & Huhtaniemi, 2009; Riggs et al., 1982). There are methods in use that increase serum T without T administration, including hCG treatment in the case of men with secondary hypogonadism. In individuals with primary hypogonadism, aromatase inhibitors can increase T-to-estradiol ratios, particularly in men with severe infertility, but often this approach is ineffective (Perheentupa & Huhtaniemi, 2009).

Increasing intratesticular and serum T by stimulating the Leydig cells themselves could have great advantages. Such an approach should not elicit significant fluctuations in T levels because T formation would be regulated at least in part by the negative feedback of T on LH. Nor would there be T transfer to others via contact. Fertility should be preserved, not suppressed, because the hypothalamic-pituitary axis should not be shut down as with exogenous T. Indeed, the local stimulation of Leydig cell T production by the use of drug ligands that target proteins involved in cholesterol import into mitochondria, the rate-determining step in T formation, might actually support or enhance spermatogenesis because intratesticular T levels should increase, not decrease. This advantage could be of great benefit to the many young men with primary hypogonadism who wish to father children. If successful, T replacement through the administration of drugs targeting proteins involved in T formation could constitute a paradigm shift in the treatment of hypogonadal men.

In summary, the design of new therapies that increase intratesticular bioactive androgen levels without affecting the hypothalamic-pituitary axis would be of great benefit to numerous patients. This approach requires an understanding of the series of steps leading to T formation and how they are regulated, and the identification of key steps amenable to pharmacological modulation to induce T.




4 STEROID BIOSYNTHESIS

Steroidogenic cells are defined by their ability to convert the precursor cholesterol to pregnenolone in the mitochondrial matrix through the function of the cytochrome P450 side-chain cleavage enzyme (CYP11A1) (Jefcoate, 2002), which is a member of a large family of P450 enzymes. Seven members of this family are targeted to mitochondria and the remaining 50 members to the endoplasmic reticulum (ER). Six of the members of this family are involved in steroidogenesis. Mitochondrial CYP11A1 converts cholesterol to pregnenolone, with the latter, then converted to progesterone by 3β-hydroxysteroid dehydrogenase (3βHSD; HSD3B) present in both the mitochondria and ER. CYP17A1, present in the ER, possesses both 17α-hydroxylase and 17,20-lyase activities. In the gonads of men, CYP17A1 converts progesterone to dehydroepiandrosterone which is further metabolized to T by 17-βHSD (Fig. 1).

Figure 1 Summary of the steroidogenic pathway in the testis. The steroid hormone biosynthesis pathway in the Leydig cells of the testis is shown in this schematic representation. Cholesterol is the sole precursor of all steroids and it is through several enzymatic reactions in mitochondria, cytoplasm, and ER that different types of steroids are synthesized due to the presence and activity of specific enzymes. P5, pregnenolone; P4, progesterone; T, testosterone.




The rate-limiting step in steroidogenesis is the import of cholesterol into mitochondria to become accessible to CYP11A1. By limiting access of the hydrophobic cholesterol molecule to CYP11A1, steroidogenic cells are able to control the amount of steroids they produce (Rone, Fan, & Papadopoulos, 2009). Steroid hormones are continuously synthesized. However, in response to increased circulating peptide hormones (LH in the case of Leydig cells), the rate of steroid hormone synthesis is greatly increased. LH BINDS ITS COGNATE RECEPTORS ON THE SURFACE OF LEYDIG CELLS AND STIMULATES INTRACELLULAR SIGNALING CASCADES, OF WHICH THE cAMP PATHWAY, THROUGH PROTEIN KINASE A (PKA), IS THE MOST PROMINENT. ACUTELY, THESE EVENTS STIMULATE THE IMPORT OF CHOLESTEROL TO THE OMM WHERE IT IS CONVERTED TO PREGNENOLONE BY CYP11A1 (Jefcoate, 2002; Papadopoulos, Liu, & Culty, 2007). IN ADDITION, HORMONAL AND cAMP STIMULATION OF STEROIDOGENIC CELLS ARE IMPORTANT FOR THE CHRONIC REGULATION OF STEROIDOGENESIS, AS CONTINUED STIMULATION IS NECESSARY TO ENSURE PROPER EXPRESSION LEVELS OF STEROIDOGENIC PROTEINS AND ENZYMES AS WELL AS STEROIDOGENIC METABOLIC FLUX (Simpson & Waterman, 1988).

The mitochondrion is a double-membrane organelle with an aqueous intermembrane space (IMS) between OMM and IMM, where CYP11A1 resides. Cholesterol molecules are hydrophobic. Thus, the movement of cholesterol across the aqueous microenvironment of the mitochondria requires the involvement of intracellular machinery (Mesmin & Maxfield, 2009). Indeed, steroidogenic cells possess a multicomponent protein machine, the transduceosome, an ensemble of cytoplasmic and resident mitochondrial proteins that receive hormonal signals and is involved in the translocation of cholesterol across the IMS at contact sites between the OMM and IMM (Papadopoulos et al., 2007; Rone, Liu, et al., 2009) (Fig. 2). The transduceosome OMM proteins were identified as the 30-kDa voltage-dependent anion channel 1 (VDAC1) and the 18-kDa translocator protein TSPO, a high-affinity drug- and cholesterol-binding protein previously named the peripheral-type benzodiazepine receptor (McEnery, Snowman, Trifiletti, & Snyder, 1992; Papadopoulos et al., 2006). In addition to the mitochondrial proteins making up core structural and enzymatic components of the transduceosome, cytoplasmic proteins are instrumental in regulating its assembly and cholesterol transport activity. Hormonal stimulation was found to promote the clustering of TSPO, which was correlated with steroidogenesis and could be suppressed by the PKA inhibitor H-89 (Boujrad, Gaillard, Garnier, & Papadopoulos, 1994). This suggests an active role for PKA in the assembly of the transduceosome. Yeast two-hybrid screening for additional cellular partners of TSPO yielded the acyl-CoAbinding domain family protein ACBD3/PAP7 (Li, Degenhardt, et al., 2001), which serves to scaffold the cytosolic PKA-RI subunits to the transduceosome (Li, Degenhardt, et al., 2001; Liu, Rone, & Papadopoulos, 2006). An additional ACBD protein family member, ACBD1, participates in transduceosome function. Originally identified through its ability to displace benzodiazepine bound to GABA receptor sites in neurons (Costa & Guidotti, 1991), and hence originally named the diazepam-binding inhibitor, ACBD1 acts on TSPO and stimulates steroidogenesis (Boujrad et al., 1994; Papadopoulos, Berkovich, Krueger, Costa, & Guidotti, 1991).

While gene expression of the transduceosome components has been the focus of numerous studies, much less is known about mechanisms through which transduceosome components are targeted to their proper locations in the mitochondria, a factor critical to their function in steroidogenesis (Black, Harikrishna, Szklarz, & Miller, 1994).


4.1 Mitochondrial protein import and chaperones

4.2 Transduceosome
4.2.1 Translocator protein
4.2.2 Voltage-dependent anion channel 1
4.2.3 Steroidogenic acute regulatory protein
4.2.4 Protein kinase A
4.2.5 Acetyl CoA-binding domain 3

4.3 Metabolon
4.3.1 Cholesterol side-chain cleavage cytochrome P450 (CYP11A1)
4.3.2 AAA+ ATPase, ATAD3


5. CAN SERUM TESTOSTERONE LEVELS BE INCREASED BY STIMULATING THE LEYDIG CELLS THEMSELVES?

As indicated above, there are good reasons to increase serum T levels in hypogonadal men, but doing so by TRT is less than ideal for both young and aging men. There are methods to increase serum T without T administration, including administering LH (or hCG) for men with secondary hypogonadism. In men with primary hypogonadism, aromatase inhibitors can increase T-to-estradiol ratios, but often this approach is ineffective (Perheentupa & Huhtaniemi, 2009). Increasing intratesticular and serum T by stimulating the Leydig cells themselves could have great advantages in part because doing so would be expected to result in the physiological regulation of the T that is produced by its negative feedback on LH. This is, in contrast, to essentially flooding the system with T, as with exogenous T administration. Stimulating the Leydig cells should not elicit significant fluctuations in T levels, and nor there be T transfer to others via contact. Fertility should be preserved, not suppressed, because the hypothalamic-pituitary axis should not be shut down as with exogenous T. Indeed, the local stimulation of Leydig cell T production might actually enhance spermatogenesis because intratesticular T levels should increase, not decrease. Moreover, Leydig cells produce more than just T, and so their stimulation might produce Leydig cell products in addition to T that might have effects on spermatogenesis. With an understanding of the series of steps leading to T formation derived from numerous studies conducted over the years and the identification of key steps that have been shown to be amenable to pharmacological modulation so as to induce T production, it now might be possible to stimulate Leydig cells to produce additional T as a means by which to increase serum T levels.


5.1 TSPO drug ligands

5.2 14-3-3γ and ε proteins




6. CONCLUSION

Cholesterol is the main precursor of T. Its import into mitochondria is the rate-limiting step in steroidogenesis. After a decade of intensive research, it is now clear that cholesterol import into the mitochondria in response to hormonal stimulation is carried out through a multiprotein complex called transduceosome, and not by individual proteins (Midzak, Rone, Aghazadeh, Culty, & Papadopoulos, 2011; Papadopoulos & Miller, 2012; Rone et al., 2012).

In search of mechanisms inducing T production in the Leydig cell as an alternative to TRT, components of the transduceosome were identified to be critical for T formation and “druggable” candidates. Indeed, TSPO and 14-3-3ε–VDAC1 interactions were shown to be viable targets which, when activated, result in increased T formation both in vitro and in vivo, thus providing new means for the treatment of androgen deficiency in hypogonadal men.

 

[Nelson Vergel]

After 3 weeks of taking 100 mg per night of pregnenolone. Here are my results (LC/MS). Still waiting for progesterone LC/MS. It went from undetectable to 95 ng/dL. I am going to increase dose to 150 mg per night.

 

[Anonymon]

After 3 weeks of taking 100 mg per night of pregnenolone. Here are my results (LC/MS). Still waiting for progesterone LC/MS. It went from undetectable to 95 ng/dL. I am going to increase dose to 150 mg per night.

View attachment 15230

And you’ve still had no issues with sleep or anything like that? And still aren’t on HCG right now?

 

[Fortunate]

After 3 weeks of taking 100 mg per night of pregnenolone. Here are my results (LC/MS). Still waiting for progesterone LC/MS. It went from undetectable to 95 ng/dL. I am going to increase dose to 150 mg per night.

View attachment 15230

I got labs drawn today. Will post when I get them.

 

[Gianluca]

@Gianluca, thanks for the information. I have watched most of Dave Lee's two presentations on Pregnenolone and DHEA. I have to admit I am getting caught in analysis paralysis. I am not sure what to make of the information out there and how to proceed. For example, I have noticed very definite fatigue and sleepiness when taking DHEA. I have also noticed it darkness my mood. Yet, he purports DHEA does the exact opposite: take it in the morning, as it gives you energy and it is a strong antidepressant. He does provide the disclaimer that both DHEA and Pregnenolone can have opposite effects in different individuals, but I have a hard time making sense of it. He also says Pregnenolone has a sedative affect, but if you do a little mini experiment on Amazon and read user reviews of OTC Pregnenolone, many say the opposite: Be careful, it amps you up!

I have a particular interest in this because I have previously experienced "The Wall" - after doing very well on Xyosted for months, I started feeling anxious, overstimulated, overall bad. I am now back on Xyosted and doing fine, but would like something to combat that feeling if it happens again. This time, I am trying it without hCG (I am still undecided on whether or not hCG causes me problems). However, if I hit the Wall again, it would be nice to know there are options to address it. I have been around long enough to know there are typically no silver bullets out there for any one problem. But, I became intrigued when I saw Pregnenolone being discussed specifically as a way to address the anxiety issues one gets after a while on TRT (The Wall).

In any case, can you give your thoughts time of day? Do you use DHEA in the morning or evening? Have you noticed any sedation? I see you are taking Pregnenolone in the AM as both topical and oral from Empower. Have you ever tried an OTC version?

Thanks again for your thoughts and would love to hear others' experience.

if DHEA makes you drowsy, simply take it before bed. My GF takes it at night and she sleeps deeper and wake up less during the night, if I take it at night it keeps my brain active. You have to try and see how things work for you, be patient and don't over think things, I have done this big mistake many times with HRT, and still do!

not sure what Xyosted is, but start by testing your Preogesterone/DHEA, if you have the extra $ include Pregnenolone, don't take hormones if you don't need to

I see a sedative effect from Progesterone capsule, Pregnenolone is calming for me, but not really sedative.

I used Nutricology brand in the past, the only OTC I used. The same brand was recommended by Dave Lee, but I have never checked any levels after supplementing with that. My concern with that brand, is that the capsule is in a lipid matrix, which should by pass the liver, you want the opposite especially if you are dealing with anxiety, as Pregnenolone when taken orally, because the actual first pass into the liver, will get converted into Progesterone and Allopregnanolone, all good things we need as calming/anxiolytic agent.