雄蚕益肾方对雄性迟发性性腺功能减退大鼠睾丸组织生物钟基因表达的影响  

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

作　　者：吴悔 宁港 李波男 石若冰 彭阿建 王浩宇 曾顺 周兴[1] WU Hui;NING Gang;LI Bo-nan;SHI Ruo-bing;PENG A-jian;WANG Hao-yu;ZENG Shun;ZHOU Xing(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;Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Hunan Provincial Hospital of Chinese Medicine,Changsha,Hunan 410005,China)

机构地区：[1]湖南中医药大学第一附属医院男科,湖南长沙410007 [2]湖南中医药大学研究生院,湖南长沙410208 [3]湖南中医药大学,湖南长沙410208 [4]湖南省中医院,湖南长沙410005

出　　处：《中华男科学杂志》2023年第1期76-82,共7页National Journal of Andrology

基　　金：国家自然科学基金(82074444);湖南省自然杰出青年科学基金(2023JJ10032);湖南省教育厅重点项目(20A368);长沙市杰出创新青年培养计划(kq1802015);湖南中医药大学中西医结合一流学科开放基金(2020ZXYJH64);湖南中医药大学研究生创新课题(2021CX69)。

摘　　要：目的:探讨雄蚕益肾方对雄性迟发性性腺功能减退症(LOH)大鼠睾丸组织细胞生物钟基因表达的影响。方法:48只8周龄雄性SD大鼠随机分为正常组,模型组,丙酸睾酮组,雄蚕益肾方低、中、高剂量组。模型组,丙酸睾酮组,雄蚕益肾方低、中、高剂量组大鼠腹腔注射480 mg/(kg·d)D-半乳糖(D-gal),连续注射56 d进行LOH造模,正常组大鼠每日腹腔注射相同体积生理盐水。造模成功后,雄蚕益肾方低、中、高剂量组分别给予10.4、20.8、41.6 g/(kg·d)体重剂量的雄蚕益肾方中药汤剂灌胃,每组每日均早、晚各灌胃1次;丙酸睾酮组大鼠肌注5.21 mg/(kg·d)体重剂量的丙酸睾酮,每周3次;正常组和模型组给予相同体积的蒸馏水灌胃;各组均连续操作28d。实验结束后,采血离心取上清液ELISA法检测睾酮(T)水平,取睾丸组织进行RT-qPCR、Western印迹分别检测BMAL1、NR1D1、PER2、CRY1、StAR、CYP11A1 mRNA表达量和蛋白表达水平。结果:模型组大鼠血清睾酮水平、睾丸组织BMAL1、NR1D1、PER2、CRY1、StAR、CYP11A1在mRNA表达量和蛋白表达水平均显著低于正常组(P<0.05);与模型组相比,丙酸睾酮组、雄蚕益肾方低、中、高剂量组大鼠血清睾酮显著升高(P<0.05),雄蚕益肾方高剂量组大鼠睾丸组织StAR mRNA表达量、CYP11A1 mRNA表达量和蛋白表达水平显著升高(P<0.05),雄蚕益肾方低、中、高剂量组大鼠睾丸组织StAR蛋白表达水平显著升高(P<0.05),丙酸睾酮组、雄蚕益肾方低、中、高剂量组大鼠睾丸组织BMAL1、NR1D1蛋白表达水平及PER2蛋白表达水平和mRNA表达量显著升高(P<0.05),雄蚕益肾方中、高剂量组大鼠睾丸组织BMAL1 mRNA表达量、CRY1蛋白表达水平显著升高(P<0.05),雄蚕益肾方低、中、高剂量组大鼠睾丸组织NR1D1 mRNA表达量显著升高(P<0.05),丙酸睾酮组、雄蚕益肾方中、高剂量组大鼠睾丸组织CRY1 mRNA表达量显著升高(P<0.05)。结论:雄蚕益肾方能上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 i

关 键 词：雄蚕益肾方 男性迟发性性腺功能减退症 LEYDIG细胞 生物钟基因 

分 类 号：R698[医药卫生—泌尿科学]