OBJECTIVE: To analyze the correlation of serum progesterone (PROG) level with blood biochemical parameters and common traditional Chinese medicine (TCM) syndromes in male patients with type 2 diabetes mellitus (T2DM). METHODS: We collected the clinical data of 192 male patients with T2DM, who were admitted in the Department of Endocrinology, Nanjing Hospital of Chinese Medical Affiliated to Nanjing University of Chinese Medicine between January, 2018 and March, 2019. The general clinical data, C-peptide level, blood glucose level, glycated hemoglobin (HbA1c), HOMA, blood lipid level, and sex hormones were compared between the patients with normal PROG and elevated PROG levels and also between the patients with two common TCM syndromes, namely and deficiency syndrome and damp- heat accumulation in the spleen syndrome. We further compared the sex hormones, C-peptide level, HOMA, HbA1c, and blood glucose level among the patients with the two TCM syndromes having normal or elevated PROG levels. RESULTS: Compared with those in patients with normal PROG level, BMI, C-peptide, HOMA-β, and HOMA2-IR were significantly lowered and HOMA-IS, E2, and T were significantly increased in patients with elevated PROG level; no statistical differences were found in age, disease duration, waist-to-hip ratio (WHR), smoking history, blood pressure, blood glucose, blood lipids, HbA1c, LH, FSH or PRL between the two groups. Compared with the patients with damp-heat accumulation syndrome group, the patients with and deficiency syndrome were older and had a longer disease duration, a greater BMI, and higher levels of PROG, C-Peptide, HOMA-β, HOMA2-IR and HOMA-IS, but the smoking history, WHR, HbA1c, blood glucose, and sex hormone levels were comparable between the two groups. Among the 4 groups of patients with different PROG levels and TCM syndromes, significant differences were found in the levels of C-peptide, HOMA-β, HOMA-IS, HOMA2-IR, PROG, E2, T, LH and FSH, and the patients with and deficiency syndrome as well as an elevated PROG level had the lowest C-peptide level, HOMA-β and HOMA2-IR and the highest HOMA-IS, PROG, E2, T, LH and FSH. CONCLUSIONS An elevated PROG level is closely related to islet cell dysfunction and TCM syndrome types in male patients with T2DM.
Blood Glucose ; Diabetes Mellitus, Type 2 ; blood ; therapy ; Humans ; Male ; Medicine, Chinese Traditional ; Progesterone ; biosynthesis ; Syndrome ; Yin Deficiency

OBJECTIVETo investigate the expression of 17 beta-hydroxysteroid dehydrogenase type 1 (17β-HSD1) in the kidney of rats and explore the capacity of the kidney for synthesizing sex hormones.

METHODSThe expressions of 17-HSD1 and sex hormones were detected by Western blotting and radioimmunoassay in rat renal cells in primary cultured for 24 and 48 h in the presence or absence of follicle-stimulating hormone (FSH) and luteinizing hormone (LH).

RESULTSAfter cell culture for 24 h, the primary rat renal cells expressed a low level of 17β-HSD1 (0.1843±0.076), which increased to 1.6651±0.044 (P<0.01) in response to co-stimulation by FSH and LH. Low levels of estradiol, progesterone and testosterone were also detected in rat renal cells (3.30±3.78, 62.60±12.33, and 22.12±3.36, respectively), and co-stimulation of FSH and LH significantly increased their levels to 8.50±2.64, 117.80±9.79, and 45.04±4.39, respectively (P<0.05). The levels of these hormones showed no significant differences between cells cultured for 24 h and 48 h (P>0.05).

CONCLUSIONThe rat renal cells express 17β-HSD1 and are capable of stably secreting sex hormones in response to co-stimulation with FSH and LH, suggesting the capacity of the rat kidneys for synthesizing sex hormones. These findings enrich the understanding of the endocrine function of the kidney.

17-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Cells, Cultured ; Estradiol ; biosynthesis ; Follicle Stimulating Hormone ; pharmacology ; Kidney ; enzymology ; Luteinizing Hormone ; pharmacology ; Progesterone ; biosynthesis ; Rats ; Testosterone ; biosynthesis

The steroidal enzyme cytochrome P45017alpha catalyzes the conversion of progesterone and pregnenolone into androgens, androstenedione and dehydroepiandrosterone, respectively, the direct precursors of estrogens and testosterone. Dihydrotestosterone is the principal active androgen in the prostate, testosterone is also an active stimulant of the growth of prostatic cancer tissue. Inhibition of this enzyme as a mechanism for inhibiting androgen biosynthesis could be a worthwhile therapeutic strategy for the treatment of PCA. In this paper, four categories of steroidal inhibitors of cytochrome P45017alpha will be reviewed, a diverse range of steroidal inhibitors had been synthesized and shown to be potent inhibitors of P45017alpha.
Androstenedione ; biosynthesis ; Androstenes ; Androstenols ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Dehydroepiandrosterone ; biosynthesis ; Dihydrotestosterone ; metabolism ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Male ; Molecular Structure ; Pregnenolone ; metabolism ; Progesterone ; metabolism ; Prostatic Neoplasms ; pathology ; Steroid 17-alpha-Hydroxylase ; antagonists & inhibitors ; Testosterone ; biosynthesis

Animals ; Animals, Newborn ; Hypoxia-Ischemia, Brain ; metabolism ; Neuroprotective Agents ; pharmacology ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo study the role of extracellular signal-regulated protein kinase 5 (ERK5) during the biosynthesis of follicle-stimulating hormone (FSH)-mediated progesterone in primary granulosa cells.

METHODSThe expressions of phosphorylated and general forms of ERKS in primary granulosa cells after the treatment of FSH were detected by Western blot analysis. The subcellular localization of ERK5 was observed by confocal microscopy. The effect of ERK5 on FSH-mediated progesterone biosynthesis in primary granulosa cells was analyzed using recombinant adenovirus vectors.

RESULTSERK5 activation was induced by FSH in a time-dependent manner in primary cultured granulosa cells, although the general ERK5 protein level decreased also in a time-dependent manner. The treatment of FSH showed no remarkable effect on the subcellular distribution of endogenous ERK5, which was mainly in the cytoplasm of granulosa cells. The co-infection of Ad-caMEK5 and Ad-wtERK5 increased the progesterone production and StAR expression in primary cultured granulosa cells, whereas inhibition of ERK5 activation suppressed the FSH-stimulated progesterone production.

CONCLUSIONERK5 may stimulate FSH-mediated progesterone production in primary cultured granulosa cells.

Animals ; Cells, Cultured ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; drug effects ; metabolism ; Mitogen-Activated Protein Kinase 7 ; metabolism ; physiology ; Progesterone ; biosynthesis ; Rats ; Rats, Sprague-Dawley

Steroid cell tumor, not otherwise specified (NOS), are rare ovarian tumor, in addition, it is more rare in children. The majority of these tumors produce several steroid hormones, particularly testosterone. Estrogen also secreted by steroid cell tumor, NOS, but it is uncommon. Furthermore, hypertension is an infrequent sign in steroid cell tumor, NOS. An 8.5-yr-old girl with hypertension and frequent vaginal spotting visited at our clinic. On laboratory evaluation, secondary hypertension due to an elevated plasma renin level and isosexual pseudoprecocious puberty was diagnosed. Right solid ovarian mass was detected in radiologic tests. She underwent a right ooporectomy and it revealed renin and progesterone receptor positive steroid cell tumor, NOS. After operation, her blood pressure returned to normal level and vaginal bleeding disappeared. Even though this case is very rare, when hypertension coincides with virilization or feminization, a renin-secreting ovarian steroid cell tumor, NOS, should be considered.
Child ; Female ; Humans ; Hypertension/*etiology ; Ovarian Neoplasms/complications/*diagnosis/pathology ; Puberty, Precocious/enzymology/*etiology ; Receptors, Cell Surface/metabolism ; Receptors, Progesterone/metabolism ; Renin/blood ; Sex Cord-Gonadal Stromal Tumors/complications/*diagnosis/pathology ; Steroids/biosynthesis ; Tomography, X-Ray Computed ; Vacuolar Proton-Translocating ATPases/metabolism

OBJECTIVETo establish a bioluminescent MDA-MB-231 cell line which can stably express luciferase and green fluorescent protein to allow bioluminescent imaging in nude mouse models bearing human triple-negative breast cancer xenografts.

METHODSThe lentivirus carrying luc2, eGFP and neo fusion genes were packaged in 293T cells via calcium phosphate co-precipitation. Human triple-negative breast cancer cell line MDA-MB-231 was infected by the lentivirus, and the positive cell clones were tested for eGFP and luc2 expressions by fluorescence microscopy and Xenogen IVIS200 bioluminescent imaging system, respectively. MTT assay, transwell invasion assay and wound healing assay were performed to evaluate the changes in the proliferation, invasion and migration abilities of the infected cells. The cells were then orthotopically implanted into the right second mammary fat pat of female BALB/c nude mice. The tumor growth was monitored by the in vivo imaging system every week, and the tumor tissues were harvested to evaluate the in vivo stability and tumorigenicity of the modified cells using cryosection and HE staining.

RESULTSThe lentivirus-infected MDA-MB-231cells could stably express luc2 and eGFP, and the luciferase activity reached 9689 phontons/s/per cell. No significant changes occurred in the biological activities of the lentivirus-infected MDA-MB-231 cells. We successfully established the nude mouse model bearing orthotopically implanted human triple-negative breast cancer cells.

CONCLUSIONThe modified MDA-MB-231 cell line can be detected sensitively at the primary implantation site and distant metastasis site in nude mice, which provides a convenient and sensitive platform for the research of metastatic mechanism and new antitumor drugs of human triple-negative breast cancer. The combination of eGFP and luc2 is superior to single reporter gene.

Animals ; Brain Neoplasms ; secondary ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Genes, Reporter ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Luciferases ; biosynthesis ; genetics ; Luminescent Measurements ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

Effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on in vitro survival, growth, steroidogenesis, and apoptosis of buffalo preantral follicles (PFs) was investigated. PFs (200~250 microm) were isolated by micro-dissection and cultured in 0 (control), 10(-3), 10(-5), 10(-7), and 10(-9) M SNP. To examine the reversible effect of SNP, PFs were cultured with 10(-5) M SNP + 1 mM Nomega-nitro-L-arginine methyl ester (L-NAME) or 1.0 microg hemoglobin (Hb). The results showed that greater concentrations of SNP (10(-3), 10(-5), 10(-7) M) inhibited (p < 0.05) FSH-induced survival, growth, antrum formation, estradiol production, and oocyte apoptosis in a dose-dependent manner. However, a lower dose of SNP (10(-9) M) significantly stimulated (p < 0.05) the survival, growth, antrum formation, follicular oocyte maturation, and stimulated progesterone secretion compared to the control. A combination of SNP + L-NAME promoted the inhibitor effect of SNP while a SNP + Hb combination reversed this effect. Nitrate and nitrite concentrations in the culture medium increased (p < 0.05) in a dose-dependent manner according to SNP concentration in the culture medium. At higher concentrations, SNP had a cytotoxic effect leading to follicular oocyte apoptosis whereas lower concentrations have stimulatory effects. In conclusion, NO exerts a dual effect on its development of buffalo PFs depending on the concentration in the culture medium.
Animals ; *Apoptosis ; Buffaloes/*physiology ; Estradiol/biosynthesis ; Female ; Follicle Stimulating Hormone/metabolism ; NG-Nitroarginine Methyl Ester/pharmacology ; Nitrates/pharmacology ; Nitric Oxide/*metabolism ; Nitric Oxide Donors/pharmacology ; Nitrites/pharmacology ; Nitroprusside/pharmacology ; Oocytes/cytology/drug effects/growth & development/metabolism ; Ovarian Follicle/*cytology/drug effects/growth & development/*metabolism ; Progesterone/biosynthesis

OBJECTIVETo investigate the functional and metabolic alterations in cultured insulin resistant ovary model in vitro, and to observe the effect of berberine (Ber, a Chinese medical monomer) in improving insulin resistance (IR).

METHODSOvary of mouse was cultured in vitro and treated by dexamethasone (Dex) to induce IR for establishing IR model ovary. The functional alteration in model ovary was assessed through detecting glucose and hormone levels in medium using RT-PCR, meanwhile, the expression of key molecules in insulin signal and steroid synthetic pathway were detected, and condition of IR improved by berberine was evaluated also.

RESULTS(1) The model ovary was made by Dex in dose- and acting time-dependent manner. After being treated by 300 nmol/L Dex for 48 h, the glucose uptake of ovary reduced from 9.05 +/- 0.75 mg/g to 2.48 +/- 0.29 mg/g (P < 0.05); it further decreased (from 9.59 +/- 1.74 mg/g to 1.94 +/- 0.19 mg/g, P < 0.01) under the stimulation of insulin, which proved that the IR model ovary was made successfully. Berberine significantly increased the glucose uptake of model ovaries (1.89 +/- 0.33 mg/g to 13.95 +/- 3.30 mg/g, P < 0.05). (2) As compared with control group, levels of testosterone (T) and androstenedione (A2) were higher, and levels of progesterone (P) and 17-hydroxyprogesterone (17-OHP) were lower significantly in the model. Berberine reversed the alternations of T, A2 and 17-OHP levels, but did not influence the level of P. (3) RT-PCR showed that the mRNA expressions of cytochrome 17-hydroxylase (CYP17) and mini-chromosome maintenance protein-2 (MCM-2) elevated, but extracellular regulated protein-1 (ERK-1), protein kinase B (AKT-2) and glycogen synthase kinase-3 (GSK-3beta) lowered in the medium after Dex inducing. Berberine treatment restored these molecular index obviously.

CONCLUSIONS(1) Dex could induce IR in mouse ovary, which might enhance the androgenic synthesis. (2) Berberine could alleviate the degree of IR and the androgen synthesis, indicating that the Chinese sensitizing agents has favorable therapeutic effect for the treatment of polycystic ovaries.

17-alpha-Hydroxyprogesterone ; metabolism ; Androstenedione ; biosynthesis ; Animals ; Berberine ; pharmacology ; Female ; In Vitro Techniques ; Insulin ; metabolism ; Insulin Resistance ; Mice ; Mice, Inbred Strains ; Ovary ; drug effects ; metabolism ; Polycystic Ovary Syndrome ; Progesterone ; biosynthesis ; Testosterone ; biosynthesis

Animals ; Cadmium ; toxicity ; Cytochrome P-450 Enzyme System ; blood ; Female ; Ovary ; drug effects ; metabolism ; Phosphoproteins ; blood ; Progesterone ; biosynthesis ; Rats ; Rats, Wistar