五子衍宗方对环磷酰胺致小鼠睾丸氧化损伤的保护作用  

Protective effect and mechanism of Wuzi Yanzong Decoction on cyclophosphamide induced testicular oxidative damage in mice

作　　者：刘苗苗[1,2] 张长城[1] 贾亮亮[3] 王洪武[1] 曾晓[1] 何毓敏[1] 袁丁[1] 王婷[1] 

机构地区：[1]三峡大学医学院,湖北宜昌443002 [2]三峡大学化学与生命科学学院,湖北宜昌443002 [3]三峡大学人民医院药剂科,湖北宜昌443000

出　　处：《中成药》2013年第12期2591-2597,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(81173375)

摘　　要：目的研究五子衍宗方(枸杞子、覆盆子、菟丝子、车前子、五味子)对环磷酰胺致小鼠睾丸氧化损伤的保护作用及其机制。方法将8周龄的Balb/C小鼠随机分组。空白对照组腹腔注射生理盐水,实验组连续7 d腹腔注射环磷酰胺50 mg/kg,然后空白对照组和模型组灌胃生理盐水,五子衍宗方组灌胃五子衍宗方提取物28 d。检测各组小鼠睾丸质量、附睾质量、精子密度、精子活率、睾丸组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPX)活性及丙二醛(MDA)水平;原位末端标记法检测生精细胞凋亡程度,Real-time PCR检测睾丸组织中SOD、GPX、Bcl-2、Bax mRNA表达水平,免疫组织化学法检测睾丸组织中凋亡相关基因Bcl-2、Bax的表达情况。结果与模型组比较,五子衍宗方能显著增加模型小鼠睾丸质量、附睾质量、精子密度和活率,提高模型小鼠睾丸组织中SOD、GPX的活性、降低MDA水平,上调SOD3、GPX1、Bcl-2基因的表达,下调Bax基因的表达,上升Bcl-2/Bax蛋白表达的比例。结论五子衍宗方能够显著恢复环磷酰胺致小鼠的睾丸生殖功能损伤,其机制可能与提高睾丸内抗氧化损伤,减少生精细胞凋亡有关。AIM To investigate the protective effect and the possible mechanism of Wuzi Yanzong Decoction （ Lycii Fruictus, Rubi Fructus, Cuscutae Semen, Plantaginis Semen, Schisandrae chinensis Fructus） on cyclophos- phamide-induced testicular oxidative damage in mice. METHODS Eight week-old male Balb/C mice were as- signed. Mice in control group received intragastrically normal saline, and mice in experimental group were in-jected intragastrically with cyclophosphamide for seven days and then with Wuzi Yanzong Decoction for 28 days consecu- tively. The weight of testis, epididymis, sperm count and sperm motility rate were recorded, and the activities and expressions of superoxide dismutase （SOD） and glutathione peroxidase （GPX） and content of malondialdehyde （MDA） in testicular tissue were detected. TUNEL was used to assay the apoptosis of spermatogenic cells. The ex- pressions of apoptosis-related gene such as Bcl-2 and Bax were employed by Real-time PCR and immunohistochem- istry. RESULTS Compared with cyclophosphamide-treated group, the weight of testis, epididymis, sperm count, sperm motility rate, the activities of SOD and GPX, the mRNA expression of SOD3, GPX1 and Bcl-2, and the ex- pression of Bcl-2/Bax significantly increased, while the MDA content, and the mRNA expression of Bax significant- ly decreased in experinental group. CONCLUSION These results suggest that Wuzi Yanzong Decoction can sig- nificantly improve the impaired reproductive function induced by cyclophospamide in mice. One of the mechanisms may be related to enhancing the anti-oxidative capability in testis and reducing the apoptosis of spermatogenic cells.

关 键 词：五子衍宗方 环磷酰胺 睾丸 氧化损伤 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]