基于Nrf2/Keap1/HO-1通路的淫羊藿素对精索静脉曲张雄性大鼠生精功能的影响

Effect of Icariin Based on Nrf2/Keap1/HO-1 Pathway on Spermatogenic Function in Male Rats with Varicocele

目的分析基于Nrf2/Keap1/HO-1通路的淫羊藿素对精索静脉曲张雄性大鼠生精功能的影响。方法选择SPF级别7~8周龄雄性大鼠60只,分为模型组、空白对照组、阳性对照组、淫羊藿素低剂量组、中剂量组、高剂量组;模型组、阳性对照组、淫羊藿素低剂量组、中剂量组、高剂量组大鼠均构建精索静脉曲张模型,空白对照组使用假手术方案处理;空白对照组、模型组每天灌胃0.9%氯化钠注射液2 ml,低剂量组、中剂量组、高剂量组每天灌胃75 mg/kg、100 mg/kg及125mg/kg的淫羊藿素,阳性对照组采用54mg/kg迈之灵灌胃治疗;灌胃给药时间为8周,给药结束隔天处死大鼠,计算脏器指数,检测精子浓度、总活率、前向运动精子、精子头侧摆幅度(ALH)、精子曲线速度(VCL)、精子直线速度(VSL)、平均路径速度(VAP);检测左侧附睾生精细胞凋亡情况,大鼠睾丸组织超氧化物酶歧化酶(SOD)、乳酸脱氢酶同工酶(LDH-X)及丙二醛(MDA)水平;检测睾丸组织中Nrf2、Keap1和HO-1蛋白及m RNA表达。结果阳性对照组、低剂量组、中剂量组、高剂量组大鼠睾丸指数及附睾指数明显高于模型组(P<0.05);阳性对照组、低剂量组、中剂量组、高剂量组大鼠精子浓度、总活率、前向运动精子、ALH、VCL、VSL、VAP明显高于模型组(P<0.05);阳性对照组、低剂量组、中剂量组、高剂量组大鼠附睾生精细胞凋亡指数明显低于模型组(P<0.05);阳性对照组、低剂量组、中剂量组、高剂量组大鼠SOD、LDH-X明显高于模型组,MDA明显低于模型组,差异有统计学意义(P<0.05);阳性对照组、低剂量组、中剂量组、高剂量组大鼠Nrf2、Keap1和HO-1蛋白明显高于模型组(P<0.05);阳性对照组、低剂量组、中剂量组、高剂量组大鼠Nrf2、Keap1和HO-1m RNA明显高于模型组(P<0.05)。结论采用淫羊藿素治疗精索静脉曲张,可能通过调控Nrf2/Keap1/HO-1通路有效调节雄性大鼠生精功能。

Objective To analyze the effects of Icariin based on Nrf2/Keap1/HO-1 pathway on spermatogenic function of male rats with varicocele. Methods Sixty male rats with SPF of 7 to 8 weeks were selected and divided into model group, blank control group, positive control group, low dose icariin group, medium dose icariin group and high dose icariin group. Model group, positive control group, icariin low-dose group, medium-dose group and high-dose group constructed varicocele rat model, and blank control group was treated with sham operation scheme. The blank control group and model group were intragastrically treated with 2 ml normal saline, the low-dose, medium-dose and high-dose groups were intragastrically treated with 75 mg/kg, 100 mg/kg and 125mg/kg icariin, and the positive control group was intragastrically treated with 54 mg/kg mazirin. The rats were given ingastric administration for 8 weeks. The rats were sacrificed on the next day after the administration. The sperm concentration, total viability,forward motile sperm, sperm cephalic swing amplitude(ALH), sperm curve velocity(VCL), sperm linear velocity(VSL) and mean path velocity(VAP) were detected by organ index. The apoptosis of spermatogenic cells in left epididymis was detected, and the activity of superoxide dismutase(SOD), lactate dehydrogenase isoenzyme(LDH-X) and malondialdehyde(MDA) in testicular tissue of rats were detected. Protein and m RNA expressions of Nrf2, Keap1 and HO-1 in testicular tissue were detected. Results The testicular index and epididymal index of positive drug group, low dose group, medium dose group and high dose group were significantly higher than those of model group(P<0.05);The sperm concentration, total viability, forward motile sperm, VSL, VCL,VAP and ALH in positive drug group, low dose group, medium dose group and high dose group were significantly higher than those in model group(P<0.05);The apoptotic index of epididymal spermatogenic cells in positive drug group, low dose group, medium dose group and high dose group was significantly lower than that in model group(P<0.05);LDH-X and SOD in positive drug group, low dose group, medium dose group and high dose group were significantly higher than those in model group, and MDA was significantly lower than that in model group(P<0.05);Nrf2, Keap1 and HO-1 proteins in positive drug group, low dose group,medium dose group and high dose group were significantly higher than those in model group(P<0.05);Nrf2, Keap1 and HO-1m RNA in positive drug group, low dose group, medium dose group and high dose group were significantly higher than those in model group(P<0.05). Conclusion The treatment of varicocele with icariin may effectively regulate the spermatogenic function of male rats by regulating Nrf2/Keap1/HO-1 pathway.