菟丝子黄酮对雷公藤多苷片致生精细胞周期阻滞、凋亡及相关蛋白表达降低的影响  

Influence of flavonoids from Cuscutae Semen on cell cycle arrest, apoptosis and protein expression of spermatogenic cells induced by multi-glycoside from Tripterygium Wilfordii

作　　者：任献青[1] 郑贵珍[1] 苏杭[1] 张霞[1] 翟文生[1] 唐进法[1] 李伟霞[1] 丁樱[1] REN Xianqing, ZI-/ENG Guizhen, SU Hang, ZHANG Xia, ZHAI Wensheng, TANG Jinfa, LI Weixia, DING Ying(The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450000, Chin)

机构地区：[1]河南中医药大学第一附属医院,河南郑州450000

出　　处：《药物评价研究》2018年第1期55-60,共6页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(81273802);河南省科技攻关计划项目(132102310245)

摘　　要：目的研究菟丝子黄酮对雷公藤多苷片致大鼠睾丸生精细胞周期阻滞、凋亡及相关蛋白表达降低的影响。方法 SD雄性大鼠随机分为5组:对照组,雷公藤多苷片高、低剂量(12、9 mg/kg)组,雷公藤多苷片高、低剂量(12、9 mg/kg)+菟丝子黄酮(生药100 mg/kg)组,每组6只。每天ig给药1次,连续给药7 d,对照组大鼠ig给予等体积0.5%CMC-Na溶液;腹主动脉采血,制备含药血清。通过体外培养大鼠睾丸生精细胞,给予含药血清干预24 h,采用流式细胞仪检测生精细胞周期及凋亡率;采用Western blotting技术检测生精细胞中SCF/c-kit、C-myc和CREM蛋白表达。结果与对照组比较,雷公藤多苷片高剂量组使G0/G1期生精细胞百分率显著降低、S期细胞百分率显著升高(P<0.01),低、高剂量组均可显著升高生精细胞凋亡率(P<0.01);与等剂量雷公藤多苷片单给药组比较,配伍菟丝子黄酮后,显著降低S期细胞百分率、生精细胞凋亡率(P<0.01)。雷公藤多苷片低、高剂量组均使生精细胞中SCF/c-kit、C-myc和CREM蛋白表达显著降低(P<0.01);配伍菟丝子黄酮后显著增加蛋白表达量(P<0.01)。结论通过合理配伍菟丝子黄酮可显著改善雷公藤多苷片对生精细胞的损伤作用。Objective To investigate the influence and action mechanism of flavonoids from Cuscutae Semen（FCS） on the cell cycle, apoptosis and protein expression of spermatogenic cells induced by multi-glycoside from Tripterygium Wilfordii（GTW）. Methods Male SD rats were randomly divided into five groups： control group, GTW high and low dose（12 and 9 mg/kg） group, high and low dose（12, 9 mg/kg） of GTW ＋ FCS（drug 100 mg/kg） group, 6 rats in each group. Drugs was given once a day for 7 d, and control group was given an equal volume of 0.5% CMC-Na solution. The blood was collected from the abdominal aorta in rats of all the groups, and the serum containing the medicine was prepared. Spermatogenic cells of rats were cultured in vitro. The influence of serum of GTW and GTW ＋ FCS on cell cycle and apoptosis of spermatogenic cells were detected by flow cytometry; and the protein expression of SCF/c-kit, C-myc and CREW in spermatogenic cells induced by serum of GTW and GTW ＋ FCS were detected by western blotting method. Results The percentage of spermatogenic cells in G0/G1 phases could be obviously decreased by GTW（12 mg/kg）, the percentage of spermatogenic cells in S phase could be obviously increased by GTW（12 mg/kg）, and apoptotic rate of spermatogenic cells could be significantly increased by GTW（12 mg/kg and 9 mg/kg, P〈0.01）. After compatibility with FCS, the inhibitory effect of cell cycle and apoptosis of spermatogenic cells in GTW group could be significantly improved（P〈0.01）. The protein expression of SCF/c-kit, C-myc and CREM could be significantly decreased by GTW 12 and 9 mg/kg（P〈0.01）. After compatibilitywith FCS, the inhibitory effect of protein expression amount of spermatogenic cells in GTW group could be significantly improved（P〈0.01）. Conclusion It indicated that when GTW and its related preparations were used in clinic, the related damage of spermatogenic cells induced by GTW and its related preparations could be improved by reasonable compatibility wit

关 键 词：雷公藤多苷片 菟丝子黄酮 配伍 生精细胞 细胞周期 细胞凋亡 蛋白表达 

分 类 号：R965.1[医药卫生—药理学]