摘要
目的:探讨五子衍宗方对环磷酰胺(Cyclophosphamide,CTX)致成年雄性小鼠睾丸生殖细胞DNA损伤及P53通路的影响,并探讨其可能的作用机制。方法:将雄性BALB/c小鼠随机分为正常对照组、模型组及五子衍宗方低、高剂量组,五子衍宗方低、高剂量组分别提前灌胃给予五子衍宗方140、280 mg/kg,其余各组灌胃给予生理盐水。从第3天开始,除正常对照组腹腔注射生理盐水外,其余各组注射CTX 50 mg/kg,隔2 d注射1次,连续10次。参照精液分析方法检测小鼠精子浓度、精子形态正常率、精子活率;单细胞凝胶电泳观察小鼠睾丸细胞DNA损伤的变化;免疫组化检测小鼠睾丸DNA损伤蛋白γ-H2AX的定位与表达变化;Western blot检测小鼠睾丸生殖细胞内DNA损伤相关蛋白γ-H2AX、Ku70及DNA损伤修复相关蛋白p-P53、P21核内表达水平。结果:与模型组比较,五子衍宗方低、高剂量均可显著升高模型组小鼠精子浓度、精子形态正常率及精子活率,显著降低彗星尾长、尾距、Olive尾矩和尾部DNA百分率及睾丸组织中γ-H2AX的阳性点数。Western blot结果表明,与模型组比较,五子衍宗方可显著降低睾丸生殖细胞核内DNA损伤相关蛋白γ-H2AX、Ku70及DNA损伤修复相关蛋白p-P53、P21的表达。结论:五子衍宗方能显著减轻CTX致小鼠睾丸生殖细胞DNA损伤,其机制可能与调控P53信号通路有关。
Objective：To investigate the effects of Wuzi Yanzong prescription on DNA damage and P53 pathways of testis in adult male mice which induced by cyclophosphamide（ CTX）, and to explore the possible mechanism. Methods：Male BALB/C mice were ran- domly divided into normal control group, model group, Wuzi Yanzong prescription low-dose group （140 mg/kg） , and Wuzi Yanzong pre- scription high-dose group （280 mg/kg） , with eight mice in each group. From 3th day, mice were injected intraperitoneally with CTX（ 50 mg/kg） once every three days for 10 times except those in normal control group. The low-dose group and the high-dose group of Wuzi Yanzong prescription were intragastrically administered with Wuzi Yanzong prescription every day. For normal control group, the same procedure was performed with intraperitoneal normal saline and intragastrically administering. The sperm concentration, the sperm nor- momorph rate and the sperm motility rate were measured reference to human semen analysis methods. The DNA damage of germ cells in testis was observed by using single cell gel electrophoresis. The localization and expression changes of DNA damage protein y-H2AX were detected by immunohistoehemical method. The nueleoprotein level of T-H2AX, Ku70, p-P53 and I〉21 in testis were examined by Western blot. Results：Compared with model control group,Wuzi Yanzong prescription significantly increased the sperm concentration, the sperm normomorph rate and the sperm motility rate, and significantly decreased in the DNA damage as evident from the parameters, including tail length, tail moment, Olive tail moment and tail DNA percentages. Wuzi Yanzong prescription significantly decreased the positive staining of T-H2AX and the expression level of y-H2AX,Ku70,p-P53 and P21 nucleoprotein in testis. Conclusion：Wuzi Yanzong prescription can effectively protect against CTX-induced testicular DNA damage through downregulating protein nucleoprotein level of T-H2AX, Ku70 by P53 signal in mice.
出处
《中药材》
CAS
北大核心
2017年第1期183-187,共5页
Journal of Chinese Medicinal Materials
基金
国家自然科学基金(81573931
81373881
81503334)
三峡大学人才科研启动基金(KJ2014B067)
