五子衍宗方对环磷酰胺致小鼠睾丸细胞DNA损伤的影响  

Effects of Wuzi Yanzong Prescription on cyclophosphamide-induced DNA damage of testis cells in mice

作　　者：赵海霞[1] 黄威峰 宋来新[1] 刘静[1] 彭奔 张长城[1] ZHAO Hai-xia HUANG Wei-feng SONG Lai-xin LIU Jing PENG Ben ZHANG Chang-cheng(Medical College of China Three Gorges University, Yichang 443002, Chin)

机构地区：[1]三峡大学医学院,湖北宜昌443002

出　　处：《中成药》2017年第3期466-470,共5页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(81573931;81373881;81503334);三峡大学人才科研启动基金(KJ2014B067)

摘　　要：目的探讨五子衍宗方(枸杞子、菟丝子、覆盆子、车前子、五味子)对环磷酰胺致成年雄性小鼠睾丸细胞DNA损伤的影响。方法将50只成年雄性Balb/C小鼠中40只腹腔注射环磷酰胺后随机均分为模型对照组(生理盐水)以及五子衍宗方低、中、高剂量组(100、200、400 mg/kg),10只另设正常对照组(生理盐水),乙醚麻醉小鼠后颈椎脱臼处死。检测小鼠精子数量、精子活率及精子畸形率,酶联免疫吸附测定法(ELISA)法检测血清中8-羟基脱氧鸟苷(8-OHdG)的含有量,单细胞凝胶电泳技术检测睾丸细胞DNA损伤程度,Western blot检测睾丸组织中磷酸化的肿瘤抑制蛋白53(p-P53)、γ位磷酸化的组蛋白(γ-H2AX)蛋白表达。结果相较于模型对照组,五子衍宗方各组可显著升高小鼠精子数量、精子活率,显著降低精子畸形率、血清8-OHdG水平,显著降低睾丸组织中p-P53与γ-H2AX蛋白表达。结论五子衍宗方能通过下调p-P53、γ-H2AX蛋白表达来显著减轻环磷酰胺致小鼠睾丸细胞DNA损伤。AIM To investigate the effects of Wuzi Yanzong Prescription（Lycii Fructus,Cuscutae Semen,Rubi Fructus,Plantaginis Semen and Schisandrae chinensis Fructus） on DNA damage of testis cells in adult male mice induced by cyclophosphamide（CTX）.METHODS Forty out of fifty adult male Balb/C mice were injected intraperitoneally with CTX and then were randomly and equally divided into model control group（normal saline）,Wuzi Yanzong Prescription low-,medium-and high-dose groups（100,200 and 400 mg/kg）,and the other ten mice served as normal control group（normal saline）.All mice were anesthetized by inhalation of ether,and then were sacrificed by cervical dislocation.The sperm count,sperm motility and malformation rate of sperm were tested.The content of 8-hydroxy-deoxyguanosine（8-OHd G） in serum was measured using ELISA;the DNA damage degree of cells in testis was detected by single cell gel electrophoresis;the protein expressions of p-P53 andγ-H2 AX in testis were examined by Western blot.RESULTS Compared with the model control group,Wuzi Yanzong Prescription groups showed significant increase in the sperm count,sperm motility and significantly decreased malformation rate of sperm,the level of 8-OHd G in serum,and the protein expressions of p-P53 andγ-H2 AX in testis were also significantly decreased.CONCLUSION Wuzi Yanzong Prescription can significantly alleviate the DNA damage of testis cells in mice induced by CTX through down-regulating protein expressions of p-P53 and γ-H2 AX.

关 键 词：五子衍宗方 环磷酰胺 8-OHDG DNA损伤 Γ-H2AX 

分 类 号：R285.5[医药卫生—中药学]