TGF-beta1 reduces connexin43-mediated gap junctional intercellular communication in rat Leydig cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the effects of TGF-beta on the expression of connexin43 (Cx43) and Cx43-mediated gap junctional intercellular communication (GJIC) in rat Leydig cells, and investigate the association of its effects on Leydig cells with its ability of changing GJIC.</p><p><b>METHODS</b>Primarily cultured purified Leydig cells were divided into a blank control group, a positive control group (treated with the GJIC inhibitor Carbenoxolone), and four TGF-beta1 groups (treated with TGF-beta1 at the concentration of 1, 2, 5 and 10 ng/ml, respectively, for 20 hours). The localization and expression of Cx43 were detected by immunofluorescence and Western blot, and the changes in GJIC analyzed by FRAP assay.</p><p><b>RESULTS</b>Cx43 was expressed as scattered bright spots in the cytoplasm and membrane of Leydig cells. TGF-beta1 significantly elevated the expression of Cx43 in the cytoplasm, but caused no evident change in the membrane. Western blot showed an evident increase in the phosphorylation of Cx43 with the increased concentration of TGF-beta1 as compared with that of the blank control group (P < 0.05). After 20 hours of treatment with TGF-beta1 at 5 ng/ml, the fluorescence intensity of Leydig cells was markedly reduced (P < 0.01), with a mean fluorescence recovery rate of merely (43.58 +/- 1.87)%.</p><p><b>CONCLUSION</b>TGF-beta1 could significantly down-regulate GJIC between adjacent Leydig cells, and this inhibitory effect may be achieved by promoting the expression of Cx43 in the cytoplasm and elevating the phosphorylation of Cx43.</p>

Effects of Wenyang Shengjing Decoction containing serum on the estradiol secretion of Leydig cells of sterile rats of shen-yang deficiency / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of Wenyang Shengjing Decoction (WSD) containing serum on the estradiol (E2) secretion, the synthesized cytochrome P450 aromatase (P450arom) activities, as well as the expression of its encode gene CYP19 in Leydig cells of male sterile rats of adenine induced Shen-yang deficiency (SYD).</p><p><b>METHODS</b>Experimental rats were randomly divided into 4 groups, i.e., the normal control group, the high, middle, and low dose WSD groups, 5 in each group. The normal saline, low, middle, and high dose WSD were respectively given to rats of all groups for 10 successive days. Blood was drawn from rats' heart 2 h after the last gastrogavage. The serum was separated after centrifuge. Leydig cells isolated and purified from SYD rats were primary cultured in vitro and divided into 5 groups in random, i. e., the blank control group, the model group, the high, middle, and low dose WSD groups (1.2, 1.0, and 0.8 g/mL, respectively). The content of E2 released in the culture supernate was determined by radioimmunoassay. The P450arom activity was detected by tritium release assay. Meanwhile, the mRNA and protein expressions of CYP19 were analyzed using fluorescent quantitative PCR and Western blot respectively.</p><p><b>RESULTS</b>Compared with the blank control group, the E2 secretion of the supernate of Leydig cells obviously decreased in the model groups, accompanied with the inhibition of P450arom activities, significant decreased protein and mRNA expressions of CYP19 (P < 0.01, P < 0.05). Compared with the model group, after intervened by WSD containing serum, the E2 secretion in the Leydig cells could be significantly increased, the P450arom activities up-regulated, the CYP19 expressions up-regulated at the protein and mRNA levels partially in a dose-dependent manner (P < 0.01, P < 0.05).</p><p><b>CONCLUSIONS</b>WSD containing serum could effectively elevate the E2 secretion in Leydig cells, which might be partially achieved through up-regulating P450arom activities and enhancing the gene expression of CyP19. This might be one of its mechanisms of action for treating male infertility of SYD.</p>

Therapeutic effect on essential hypertension treated with combined therapy of acupuncture and medication / 中国针灸

<p><b>OBJECTIVE</b>To explore the mechanism of acupuncture combined with medication for treatment of essential hypertension (EH).</p><p><b>METHODS</b>Sixty cases of EH were randomly divided into a combined acupuncture and medication group (group A) and a medication group (group B), 30 cases in each one, treated with acupuncture in combination with oral administration of Felodipine, and simple oral administration of Felodipine respectively. Before and after treatment, the changes of blood pressure, and the contents of E-selectin (Es), iNOS and eNOS were determined.</p><p><b>RESULTS</b>After treatment, the blood pressure declined in either group. The total effective rate in group A was 86.7% (26/30), which was superior to that of 73.3% (22/30) in group B. After treatment, the plasma Es and iNOS contents in two groups decreased as compared with those before treatment (both P < 0.01), of which, plasma Es content in group A decreased apparently as compared with group B (P < 0.01). After treatment, the content of plasma eNOS increased as compared with that before treatment in group A (P < 0.01).</p><p><b>CONCLUSION</b>The mechanism of acupuncture on anti-blood pressure probably relies on the improvements in vascular endothelial cellular function so that Es, iNOS and eNOS expression can be recovered to normal level and ultimately blood pressure is adjusted.</p>

Regulatory effect of Wenyang Whengling Decoction on Smads expressions in testis of sterile rats with Shen-yang deficiency / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the localization and distribution of expressions of Smads (mother against dpp), the intracellular signal transduction molecules in the transforming growth factor-beta (TGF-beta) family, in the testis of male sterile rats with Shen-yang deficiency induced by adenine and to observe the effect of Wenyang Shengjing Decoction (WSD) on these expressions.</p><p><b>METHODS</b>Rats were randomly divided into the control group, the model group and the WSD group. Localization and distribution of Smad 1, Smad 2 and Smad 4 expressions were detected by immunohistochemistry SABC and semi-quantitative RT-PCR and analyzed statistically by image analysis system; the contents of testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were detected by radioimmunoassay; and the weights of body, testis and epididymis, as well as sperm number of rats were also measured.</p><p><b>RESULTS</b>Smad 1 and Smad 2 were expressed in cytoplasm of all levels of spermatogenic cells in rats' testis with their positive immuno-responsive substance locating in the cytoplasm, and positive Smad 2 expression could also be found in cytoplasm of Sertoli's cell, but both of them showed negative response in Leydig's cell; Smad 4 was positively expressed in cytoplasm of Leydig's cell but showed negative response in spermatogenic cell and Sertoli's cell. Compared with the normal control, Smad 1 expression was lower (P < 0.05), but Smad 2 and Smad 4 were higher in the model group (both P < 0.05), these abnormal changes could be reversed by WSD treatment (all P < 0.05). Compared with the normal control group, the body weight, sperm number and serum T level were lower, and levels of FSH and LH were higher (all P < 0.05) in the model group, which could all be improved by WSD (P < 0.05); the weights of testis and epididymis were unchanged in all groups (P > 0.05).</p><p><b>CONCLUSION</b>WSD could not only increase the sperm number through elevating serum T level and decreasing the levels of FSH and LH, but also by way of regulating Smads genes expression to adjust the levels of sex hormones, promote the production of sperm directly or indirectly, so as to treat male infertility.</p>

Expression of Smad1 and Smad5 in the testis of infertile rats with kidney-yang deficiency / 中华男科学杂志

<p><b>OBJECTIVES</b>To study the expression of Smad1 and Smad5 in the testis of infertile rats with adenine-modeled kidney-yang deficiency and the pathological mechanism of infertility with kidney-yang deficiency, attempting to obtain experimental evidence for the prevention and treatment of male infertility.</p><p><b>METHODS</b>Forty-eight 60 d male SD rats were divided randomly into 6 groups with 8 in each: 7 d, 14 d and 21 d kidney-yang deficiency groups, and 7 d, 14 d and 21 d control groups. The experimental rats had been fed with adenine (300 mg/kg) and the expression levels of Smad1 and Smad5 were measured with immunohistochemical SABC method at the 7th, 14th and 21st day.</p><p><b>RESULTS</b>Smad1 immunoreactivity was mainly located in the spermatogonia, spermatocytes and spermatids, and the reactive substance distributed in cytoplasm with negative nuclei. Sertoli cells and Leydig cells were negative. Compared with the control, the expression level of Smad1 was decreased significantly at the 21st day (P < 0.05), but with no significant difference at the 7th and 14th day (P > 0.05). Smad5 immunoreactivity was mainly located in the spermatogonia and spermatocytes, and the reactive substance distributed in cytoplasm with negative nuclei. Compared with the control, the expression level of Smad5 was not significantly different at the 7th day (P > 0.05). The expression of Smad5 was negative at the 14th and the 21st day.</p><p><b>CONCLUSION</b>The weaker expression of Smad1 and no expression of Smad5 may be one of the pathological mechanisms of infertility with adenine-modeled kidney-yang deficiency.</p>