[雄蚕益肾方上调迟发性性腺功能减退大鼠睾丸组织中生物钟基因的表达]

[Xiongcan Yishen Formula up-regulates the clock gene expressions in the testis tissue of the rats with late-onset hypogonadism].

作者信息

Wu Hui, Ning Gang, Li Bo-Nan, Shi Ruo-Bing, Peng A-Jian, Wang Hao-Yu, Zeng Shun, Zhou Xing

机构信息

Department of Andrology, The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China.

Graduate School, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China.

出版信息

Zhonghua Nan Ke Xue. 2023 Jan;29(1):76-82.

DOI:

PMID:37846837

Abstract

OBJECTIVE

To investigate the effect of Xiongcan Yishen Formula (XYF) on the expressions of the clock genes in the testis tissue of the rats with late-onset hypogonadism (LOH).

METHODS

Forty-eight 8-week-old male SD rats were randomly divided into 6 groups, normal control, model control, testosterone propionate (TP), and low-, medium- and high-dose XYF. The LOH model was made in the later 5 groups of rats by intraperitoneal injection of D-galactose at 480 mg/kg/d for 56 successive days, while the normal controls were injected with the same volume of normal saline. After modeling, the rats in the low-, medium- and high-dose XYF groups were treated intragastrically with XYF at 10.4, 20.8 and 41.6 g/kg/d, bid, respectively, those in the normal and model control groups with the same volume of distilled water, and those in the TP group intramuscularly with TP at 5.21 mg/kg/d, qd alt, all for 28 days. After treatment, the supernatant was obtained for measurement of the serum T level by ELISA, and the testis tissue collected for determination of the mRNA and protein expressions of BMAL1, NR1D1, PER2, CRY1, StAR and CYP11A1 by RT-qPCR and Western blot.

RESULTS

Compared with the normal controls, the rats in the LOH model control group showed significantly decreased serum T and mRNA and protein expressions of BMAL1, NR1D1, PER2, CRY1, StAR and CYP11A1 (P < 0.05). In comparison with the findings in the model controls, the T level was remarkably increased in the TP and XYF groups (P < 0.05), the expressions of StAR mRNA and CYP11A1 mRNA and protein markedly up-regulated in the high-dose XYF group (P < 0.05), and so was the expression of the StAR protein in the XYF and TP groups (P < 0.05), those of BMAL1 and NR1D1 proteins and PER2 mRNA and protein in the high-dose XYF group (P < 0.05), those of BMAL1 mRNA and CRY1 protein in the medium- and high-dose XYF groups (P < 0.05), that of NR1D1 mRNA in the XYF and TP groups (P < 0.05), and that of CRY1 mRNA in the medium- and high-dose XYF and TP groups (P < 0.05).

CONCLUSION

Xiongcan Yishen Formula could up-regulate the expressions of the clock genes in the testis tissue of the LOH rats and increase the serum T level as well, which may underlie the mechanisms of Xiongcan Yishen Formula acting on LOH.

摘要

目的

探讨雄蚕益肾方（XYF）对迟发性性腺功能减退（LOH）大鼠睾丸组织中生物钟基因表达的影响。

方法

将48只8周龄雄性SD大鼠随机分为6组，即正常对照组、模型对照组、丙酸睾酮（TP）组以及XYF低、中、高剂量组。后5组大鼠通过腹腔注射480 mg/kg/d的D-半乳糖连续56天建立LOH模型，正常对照组注射等量生理盐水。造模后，XYF低、中、高剂量组大鼠分别按10.4、20.8和41.6 g/kg/d的剂量灌胃给予XYF，每日2次；正常对照组和模型对照组大鼠灌胃给予等量蒸馏水；TP组大鼠肌肉注射TP，剂量为5.21 mg/kg/d，每日1次，均持续28天。治疗后，取上清液采用酶联免疫吸附测定法（ELISA）检测血清睾酮（T）水平，取睾丸组织采用反转录-定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测BMAL1、NR1D1、PER2、CRY1、类固醇生成急性调节蛋白（StAR）和细胞色素P450 11A1（CYP11A1）的mRNA和蛋白表达。

结果

与正常对照组相比，LOH模型对照组大鼠血清T水平以及BMAL1、NR1D1、PER2、CRY1、StAR和CYP11A1的mRNA和蛋白表达均显著降低（P<0.05）。与模型对照组相比，TP组和XYF组大鼠T水平显著升高（P<0.05）；高剂量XYF组StAR mRNA、CYP11A1 mRNA及蛋白表达显著上调（P<0.05）；XYF组和TP组StAR蛋白表达显著上调（P<0.05）；高剂量XYF组BMAL1和NR1D1蛋白以及PER2 mRNA和蛋白表达显著上调（P<0.05）；中、高剂量XYF组BMAL1 mRNA和CRY1蛋白表达显著上调（P<0.05）；XYF组和TP组NR1D1 mRNA表达显著上调（P<0.05）；中、高剂量XYF组和TP组CRY1 mRNA表达显著上调（P<0.05）。