丹参酮ⅡA通过调控p38MAPK/NF-κB信号通路减轻糖尿病雄性大鼠生殖损伤实验研究

TanshinoneⅡA alleviates reproductive damage in diabetic male rats by regulating p38MAPK/NF-κB signaling pathway

目的:探讨丹参酮ⅡA(TanⅡA)对糖尿病雄性大鼠生殖损伤的影响及其机制。方法:随机选取43只雄性SD大鼠中的35只通过高糖高脂喂养联合腹腔注射链脲佐菌素构建糖尿病动物模型,并分为模型组、TanⅡA(6 mg/kg)组、SB203580[p38丝裂原活化蛋白激酶(p38MAPK)抑制剂,2 mg/kg]组和TanⅡA+SB203580(6 mg/kg+2 mg/kg)组,每组8只;剩余8只设为正常组。给药治疗4周后,检测空腹血糖(FBG)水平。ELISA法检测血清睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH)含量。检测精子质量(浓度、存活率和活力),测量睾丸重量并计算睾丸指数。HE染色法观察睾丸组织病理学改变。检测睾丸组织总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)活力和丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-8含量。Western blot法检测睾丸组织p38MAPK、p-p38MAPK、核因子-κB p65(NF-κB p65)、p-NF-κB p65、高迁移率族蛋白B1(HMGB1)表达。结果:与模型组比较,TanⅡA组、SB203580组和TanⅡA+SB203580组FBG水平比较差异无统计学意义(均P>0.05);T、LH、FSH含量和精子浓度、存活率及活力升高(均P<0.05);睾丸重量和睾丸指数升高(均P<0.05);睾丸组织病理学改变明显改善;T-SOD、CAT活力升高且MDA、TNF-α、IL-1β、IL-8含量降低(均P<0.05);HMGB1水平和p-p38MAPK/p38MAPK、p-NF-κB p65/NF-κB p65比值降低(均P<0.05)。TanⅡA+SB203580组以上指标优于TanⅡA组和SB203580组(均P<0.05)。结论:TanⅡA可能通过抑制p38MAPK/NF-κB信号通路,降低氧化应激水平和炎症反应,从而减轻糖尿病雄性大鼠生殖损伤。

Objective:To investigate the effect of tanshinoneⅡA(TanⅡA)on reproductive damage in diabetic male rats and its mechanism.Methods:Randomly selected thirty-five out of fourty-three SD male rats were used to prepare the diabetic animal model by feeding with high glucose and fat combined with intraperitoneal injection of streptozotocin,and which were divided into the model group,TanⅡA(6 mg/kg)group,SB203580(p38 mitogen activated protein kinase[p38MAPK]inhibitor,2 mg/kg)group and TanⅡA+SB203580(6 mg/kg+2 mg/kg)group,with eight rats in each group.And another eight rats were set as normal group.After four weeks of medication,the level of fasting blood glucose(FBG)was detected.ELISA method was used to detect the content of serum testosterone(T),luteinizing hormone(LH)and follicle-stimulating hormone(FSH).The sperm quality(sperm concentration,survival rate and vitality)was analyzed.The quality of testis was weighed and calculating testicular index.HE staining was used to observe the pathological changes of testis tissue.The activity of total superoxide dismutase(T-SOD),catalase(CAT)and the content of malondialdehyde(MDA),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-8 in testicular tissue were determined.Western blot method was used to detect the expression of p38MAPK,p-p38MAPK,nuclear factor-κB p65(NF-κB p65),p-NF-κB p65,high mobility group protein B1(HMGB1).Results:Compared with the model group,there was no significant difference in FBG level of TanⅡA,SB203580 and TanⅡA+SB203580 groups(all P>0.05).The content of T,LH,FSH and the sperm concentration,survival rate,vitality were increased(all P<0.05).The testis mass and testis index were increased(all P<0.05).The pathological changes of testis tissue were improved.The activity of T-SOD,CAT and the content of MDA,TNF-α,IL-1β,IL-8 were decreased(all P<0.05).The ratio of p-p38MAPK/p38MAPK,p-NF-κB p65/NF-κB p65 and the expression of HMGB1 were decreased(all P<0.05).The effects of TanⅡA+SB203580 on the indexes were better than those of TanⅡA group and SB203580 group(all P<0.05).Conclusion:TanⅡA may reduce reproductive damage in male diabetic rats by inhibiting p38MAPK/NF-κB signaling pathway,reducing oxidative stress and inflammatory response.