淫羊藿苷改善白消安联合环磷酰胺诱导的雄性小鼠生殖功能损伤的作用研究  

Icariin improves busulfan-and cyclophosphamide-induced reproductive function damage in male mice

作　　者：吴瑶 时姗姗 柳海燕 黄皓 石兴华 侯景 WU Yao;SHI Shan-shan;LIU Hai-yan;HUANG Hao;SHI Xing-hua;HOU Jing(School of Ecology,Lishui College,Lishui,Zhejiang 323000,China;Department of Pathology,Jiangsu Province People's Hospital/The First Affiliated Hospital Nanjing Medical University,Nanjing,Jiangsu 210029,China;Suichang Forestry Development Center,Lishui,Zhejiang 323000,China)

机构地区：[1]丽水学院生态学院,浙江丽水323000 [2]江苏省人民医院病理科,江苏南京210029 [3]遂昌林业发展中心,浙江丽水323000

出　　处：《中华男科学杂志》2025年第1期25-33,共9页National Journal of Andrology

基　　金：丽水市本级重点研发计划(2021ZDYF07);丽水市公益性技术应用研究计划(2023GYX45);浙江省自然科学基金探索青年项目(LQ23C200002)。

摘　　要：目的:通过小鼠体内外实验综合评价淫羊藿苷对雄性小鼠生殖功能损伤的缓解作用。方法:体外分离KM小鼠睾丸间质细胞,CCK-8法考察淫羊藿苷对小鼠睾丸间质细胞的毒性影响;KM小鼠随机分成6组,分别为正常对照组,模型对照组(白消安10 mg/kg联合环磷酰胺120 mg/kg给药3 d),建模后淫羊藿苷低、中、高剂量给药组(淫羊藿苷灌胃20、40、80 mg/kg)以及阳性对照组(五子衍宗丸灌胃2 g/kg),连续给药30 d,生殖脏器称重计算脏器系数,并进行精子计数;HE染色观察睾丸组织病理学变化;ELISA法检测血清睾酮水平;比色法检测睾丸氧化应激指标和NO信号通路指标;qPCR法检测促凋亡基因Fas、Bax的表达情况。结果:细胞实验证实淫羊藿苷对分离的小鼠睾丸间质细胞无毒性作用,并可有效减少环磷酰胺、白消安诱导的细胞毒性作用,促进睾酮的分泌。小鼠生殖功能损伤模型结果显示,80 mg/kg的淫羊藿苷能显著提高睾丸脏器指数、促进精子生成量(P<0.05),但对附睾脏器指数影响较小。睾丸组织形态学观察发现淫羊藿苷处理后小鼠睾丸结构显著改善,生精小管界膜完整,各类生精细胞数量增多。小鼠睾丸氧化应激结果分析可知,与模型对照组比较,淫羊藿高剂量组中MDA含量显著降低(P<0.01),T-AOC、T-SOD表达量显著上升(P<0.05);同时,睾丸组织中NO含量和NOS酶活水平较模型对照组显著下降(P<0.01)。qPCR结果显示淫羊藿苷对促凋亡基因Bax和Fas的mRNA水平表达呈现出抑制作用。结论:淫羊藿苷可能通过提升睾丸抗氧化水平,降低NOS活性进而抑制NO水平,并调节白消安和环磷酰胺诱导的睾丸细胞凋亡作用,从而改善氧化应激状态对小鼠睾丸功能的损伤,发挥睾丸生精功能的保护作用。Objective:To comprehensively evaluate the effect of icariin in alleviating reproductive function damage(RFD)in male mice via in vitro and in vivo experiments.Methods:We isolated Leydig cells from 60 KM male mice in vitro,and examined the toxic effect of icariin on the Leydig cells using Cell Counting Kit-8(CCK-8).We equally randomized the mice into six groups:normal control,RFD model control(made by intraperitoneal injection of busulfan at 10 mg/kg combined with cyclophosphamide(CP)at 120 mg/kg),positive control,and low-,medium-and high-dose icariin.After modeling,we treated the mice in the positive control group with Wuziyanzong Pills and those in the low-,medium-and high-dose icariin groups by intragastrical administration of icariin at 20,40 and 80 mg/kg-1,respectively,for 30 successive days.Then we obtained the weight and visceral coefficients of the reproductive organs,calculated the sperm count,observed the pathological changes in the testis tissue by HE staining,measured the serum testosterone(T)level by ELISA,determined the indexes of testicular oxidative stress and nitric oxide(NO)signaling pathway by colorimetric assay,and detected the expression levels of the pro-apoptotic genes Fas and Bax by qRT-PCR.Results:CCK-8 assay confirmed that icariin had no toxic effect on the isolated Leydig cells of the mice,and could effectively reduce busulfan-and CP-induced cytotoxicity and promote the secretion of serum T.Icariin at 80 mg/kg significantly increased the visceral coefficient of the testis and promoted spermatogenesis(P<0.05),but had little effect on the visceral coefficient of the epididymis in the RFD model mice.Testicular histomorphometric observation revealed significantly improved testis structure,intact boundary membrane of seminiferous tubules and increased numbers of various types of spermatogenic cells of the model mice after treated with icariin.Compared with the mice in the model control group,those treated with high-dose icariin showed a significantly reduced content of malondialdehyde(MDA)(by

关 键 词：淫羊藿苷 环磷酰胺 白消安 生殖功能 氧化应激 小鼠 

分 类 号：R697.22[医药卫生—泌尿科学]