Effect of Wi-Fi microwave irradiation on testosterone synthesis in Leydig cells of male workers / 中国职业医学

OBJECTIVE: To explore the effect of occupational wireless fidelity(Wi-Fi) microwave radiation on testosterone synthesis in male workers. METHODS: A total of 51 male workers exposed to microwave radiation in Wi-Fi test station of a mobile phone manufacturer were selected as exposure group by judgment sampling method. They were divided into <2.0 years subgroup and ≥2.0 years subgroup according to the length of work years. At the same time, 30 male workers who were not exposed to occupational hazards in the same factory were selected as the control group. Serum total cholesterol level was detected by colorimetry. Serum testosterone, cyclic adenosine monophosphate(cAMP), cytochrome P450 17 A1(P450 c17), cytochrome P450 cholesterol side chain cleavage enzyme(P450 scc), levels were detected by enzyme-linked immunosorbent assay. The relative expression of P450 scc and P450 c17 mRNA in whole blood was measured by real-time quantitative polymerase chain reaction. RESULTS: The levels of serum testosterone, P450 c17 and the relative expression of P450 c17 mRNA in workers of the exposure group were lower than that in the control group(P≤0.05), and the above indexes in the sub-exposure group with work age ≥2.0 years was lower than that in the control group(P<0.05). There was no significant difference in levels of serum total cholesterol, cAMP, P450 scc and relative expression of P450 scc in whole blood among the exposed group,two subgroups and the control group(P>0.05). CONCLUSION: Long-term exposure to Wi-Fi microwave radiation can inhibit the expression of P450 c17 mRNA and the synthesis of P450 c17 protein, both are key enzymes for testosterone synthesis in male workers, thereby affecting the synthesis and secretion of testosterone.

Effect of Heshouwuyin on protein expression of steroidogenic acute regulatory protein and cytochrome P450 cholesterol side chain cleavage enzyme in Leydig cells of rats / 解剖学报

Objective To explore the effect of Heshouwuyin on the steroidogenic acute regulatory protein (StAR) and cytochrome P450 cholesterol side chain cleavage enzyme(P450scc) protein expression in Leydig cells of rats. Methods Juvenile male Wistar rat, 10-20 days old, were used to isolate the Leydig cells and cultured, and then collected them. The oxidative damage technique was used to establish Leydig cell aging model. This study had a Leydig aging model, Heshouwuyin and Shouwu pill treatments, and normal and Heshouwuyin control group. Results The intensities of immunohistichemical staining for StAR and P450scc were significantly lower in the Leydig aging model group than those in the normal group and Heshouwuyin control group (P < 0.01), and were significantly higher in the Heshouwuyin and Shouwu pill group than that in the aging model group (P < 0.05). Heshouwuyin group was better than Shouwu pill group. Conclusion Heshouwuyin can increase the protein expression of StAR and P450scc in Leydig cells, improve the synthesis of testosterone, thus delaying the senescence of Leydig cells.

Annexin A5 regulates Leydig cell testosterone production via ERK1/2 pathway / 亚洲男科学杂志(英文版)

This study was to investigate the effect of annexin A5 on testosterone secretion from primary rat Leydig cells and the underlying mechanisms. Isolated rat Leydig cells were treated with annexin A5. Testosterone production was detected by chemiluminescence assay. The protein and mRNA of Steroidogenic acute regulatory (StAR), P450scc, 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD), and 17α-hydroxylase were examined by Western blotting and semi-quantitative RT-PCR, respectively. Annexin A5 significantly stimulated testosterone secretion from rat Leydig cells in dose- and time-dependent manners and increased mRNA and protein expression of StAR, P450scc, 3β-HSD, and 17β-HSD but not 17α-hydroxylase. Annexin A5 knockdown by siRNA significantly decreased the level of testosterone and protein expression of P450scc, 3β-HSD, and 17β-HSD. The significant activation of ERK1/2 signaling was observed at 5, 10, and 30 min after annexin A5 treatment. After the pretreatment of Leydig cells with ERK inhibitor PD98059 (50 μmol l-1 ) for 20 min, the effects of annexin A5 on promoting testosterone secretion and increasing the expression of P450scc, 3β-HSD, and 17β-HSD were completely abrogated (P < 0.05). Thus, ERK1/2 signaling is involved in the roles of annexin A5 in mediating testosterone production and the expression of P450scc, 3β-HSD, and 17β-HSD in Leydig cells.

Extremely mild cholesterol side chain cleavage enzyme deficiency with infertility as the only presenting complaint / 中华内分泌代谢杂志

[Summary] All patients with cholesterol side-chain cleavage enzyme ( P450scc) deficiency that have been reported presented with early adrenal failure. Here we described a 35-year-old male presented with infertility as the only initial presenting complaint. He had received two separate surgeries to remove bilateral testicular masses. We reevaluated the resected tumors and found testicular adrenal rest tumor ( TART) pathology in the resected tumor. We profiled steroid hormones and found significantly elevated ACTH. CT scan revealed bilateral adrenal hyperplasia. Mutation screening identified compound heterozygous mutations (R353W and P432L) in the P450scc encoding gene (CYP11A1). The patient was finally diagnosed as congenital adrenal hyperplasia.

CYP11alpha (tttta)n Microsatellite Polymorphism in Korean Patients with Polycystic Ovary Syndrome / 대한불임학회지

OBJECTIVES: To investigate the distribution and functional significance of CYP11alpha (tttta)n microsatellite polymorphism in Korean patients with polycystic ovary syndrome MATERIALS AND METHODS: Analysis of CYP11alpha(tttta)n microsatellite polymorphism was carried out on DNA samples from 97 patients with polycystic ovary syndrome and 70 normal controls. Comparison were done between PCOS patients and controls concerning CYP11alpha (tttta)n microsatellite polymorphism genotype or allele frequencies. RESULTS: The most frequent allele observed in the controls was an allele with six repeats (60.7%). Significant difference in the frequency of genotype (4R (-) genotype) having no copy of four-repeat-allele were observed between PCOS patients and controls (66.0% vs 34.0%, p=0.038, OR=1.939). But no significant difference was observed in the serum levels of total testosterone or free testosterone between 4R (+) genotype and 4R (-) genotype among PCOS patients. However, hyperandrogenic PCOS patients with 4R (+) genotype showed a higher serum testosterone levels compared to controls (mean+/-S.D: 0.49+/-0.21 ng/ml vs 0.37+/-0.18 ng/ml, p=0.037). CONCLUSION: The alleleic distribution of CYP11alpha (tttta)n microsatellite polymorphism in Korean subjects were different from those reported in Caucasians. CYP11alpha (tttta)n microsatellite polymorphism was associated with polycystic ovary syndrome in the Korean population, and may play a role in the synthesis of androgens in patients with polycystic ovary syndrome.

Expression of 3b-Hydroxysteroid dehydrogenase and P450 side chain cleavage enzyme in the human uterine endometrium

The enzyme complex 3b-hydroxysteroid dehydrogenase/delta(5)-delta(4)-isomerase (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. The expression of 3beta-HSD in human uterine endometrium during the menstrual cycle and decidua was examined in an effort to understand its role during ova implantation. 3beta-HSD was weakly expressed in the glandular epithelium of the proliferative phase and moderately expressed in the glandular epithelium of secretory phase of the endometrium. In the decidua of the ectopic pregnancy, 3beta-HSD was strongly expressed. The human uterine endometrial 3beta-HSD was identified as being the same type as the placental 3beta-HSD by RT-PCR and sequence analysis. In addition to the expression of 3beta-HSD, P450scc was expressed in the decidua of the ectopic pregnancy. These results suggest that pregnenolone might be synthesized from cholesterol by P450scc de novo and then, it is converted to progesterone by 3beta-HSD in the uterine endometrium. The data implies that the endometrial 3beta-HSD can use not only the out-coming pregnenolone from the adrenal gland but also the self- made pregnenolone to produce progesterone. The de novo synthesis of progesterone in the endometrium might be a crucial factor for implantation and maintenance of pregnancy.

A Case of Congenital Lipoid Adrenal Hyperplasia

Congenital lipoid adrenal hyperplasia is the most severe form of CAH, leading to impaired production of all steroid hormones including glucocorticoids, mineralocorticoid, and sex steroids. The affected individuals are all phenotypically female with a severe salt-losing syndrome that is fatal if steroid replacement is not begun immediately after birth. The lesion of this disorder has been suggested to be in the first step of steroidogenesis of conversion of cholesterol to pregnenolone by P450scc. Recently, molecular defect of this disease has been located in the transport of cholesterol into mitochondria due to defective regulatory protein called 'steroidogenic acute regulatory protein' while the enzyme P450scc itself is normal, differing from other types of congenital adrenal hyperplasia. We experienced 2 1/2 month old phenotypical girl who was admitted due to lethargic state and persistent vomiting with severe hyperkemia and hyponatremia. Blood levels of cortisol, aldosteron, and 17-OH progesteron were low and levels of ACTH, angiotensin, and plasma renin activity were high, urinary levels of 17-KS and 17-OHCH were low. The patient was found to have karyotype of 46, XY and has been being treated with predinisolone, fluorocortisol and sodium supplement in diet and doing well. The molecular study for P450scc gene and StAR gene of patient and family is in progress.

Effects of Endurance Training and Chinese Medicine Compound for Invigorating Qi and Tonifying the Kidney on Testicular StAR and P450_(SCC) mRNA Levels in Rats / 中国运动医学杂志

Objective Effects of exercise training and Chinese Medicine Compound for Invigorating Qi and Tonifying the Kidney on testicular StAR and P450 SCC mRNA expression in rats were observed. Methods 50 Wistar rats were randomly divided into 4 groups: sedentary control group(SC, n=10), sedentary with medicine group (SM,n=10), training group(T, n=15) and training with medicine group(TM, n=15). After 6 weeks swimming training , serum testosterone level of rats were tested through radioimmunological method; using RT-PCR, we measured testicular StAR and P450 SCC mRNA levels of rats. Results (1) In T group, serum testosterone level was lower than that in SM and TM groups. Whereas, compared with SC group, the serum testosterone level in TM group kept steadiness. (2) In T group, the gene expression of StAR was markedly lower than that in SC control (P

THE REGULATORY EFFECT OF hCG ON STEROIDOGENIC ENZYMES IN ADULT MOUSE LEYDIG CELLS / 解剖学报

Objective To study the regulation role on molecular level of hCG on P450scc,P450c17 and 3?HSD in adult mouse Leydig cells. Methods Leydig cells of adult mice were cultured in vitro for 1 and 8 hours with or without hCG,then P450scc,P450c17 and I,VI total 3?HSD mRNA levels were measured respectively by the semi quantitative RT PCR as well as Scal restriction endonuclease incised methods.Meanwhile,the testosterone contents were measured in two groups. Results 1.In stimulated group Leydig cells cultured for 1 and 8 hours with hCG,the testosterone contents were higher significantly than those in control groups(1 hour P 0.05),but the ratio of type VI/I 3?HSD mRNA gradually increased. Conclusion hCG can stimulate the transcription of P450scc,P450c17 and type VI 3?HSD in adult mouse Leydig cells.