苍附导痰汤对肥胖型PCOS-IR模型大鼠卵巢TLR4/NF-κB p65信号通路的影响

Effect of cangfudaotan decoction on TLR4/NF-κB p65 signaling pathway in obese PCOS-IR rats

目的 探讨苍附导痰汤对肥胖型PCOS-IR (polycystic ovarian syndrome-insulin resistance, PCOS-IR)模型大鼠卵巢Toll受体4(TLR4)/核转录因子κB p65(NF-κB p65)信号通路的调控作用。方法 48只♀大鼠随机分为正常组8只和模型组40只。来曲唑(1 mg·kg^(-1))联合高脂饮食建立肥胖型PCOS-IR大鼠模型,快速革兰染色法观察动情周期,挑选24只模型大鼠随机分为:模型组、阳性药(二甲双胍135 mg·kg^(-1))组、苍附导痰汤高、低剂量(57.96、14.49 g·kg^(-1))组,每组各6只,药物干预21 d。观察动情周期、卵巢和子宫指数变化;血液生化仪测定空腹血糖(FBG)和血脂(甘油三酯TG和总胆固醇TC)变化;酶联免疫吸附(ELISA)法测定空腹胰岛素(FINS)水平;免疫组化法和荧光定量PCR法检测卵巢中TLR4和NF-κB p65蛋白及基因的表达。结果 与正常组比较,模型组大鼠动情周期紊乱,卵巢多囊性改变明显,FBG、TG、TC含量和FINS、HOMA-IR水平上调,卵巢中TLR4和NF-κB p65蛋白及mRNA表达均增加(P<0.05);与模型组比较,苍附导痰汤高剂量组大鼠排卵周期得到改善,卵巢多囊性改变减轻,上述指标出现明显逆转(P<0.05)。结论 苍附导痰汤能有效改善肥胖型PCOS-IR大鼠卵巢排卵和糖脂代谢功能,作用机制可能与调控卵巢TLR4/NF-κB p65信号通路有关。

Aim To explore the regulatory effect of Cangfudaotan Decoction on the ovarian Toll receptor 4(TLR4)/nuclear transcription factor kBp65(NF-κB p65)signaling pathway in obese PCOS-IR rats.Methods Forty-eight female rats were randomly divided into normal group(n=8)and model group(n=40).The obese PCOS-IR rats were established by letrozole(1 mg·kg^(-1))combined with high-fat diet,the estrous cycle changes were observed by rapid Gram staining.24 PCOS-IR model rats were randomly divided into:model group,positive drug(metformin 135 mg·kg^(-1))group,Cangfu Daotan Decoction high-dose(57.96,14.49 g·kg^(-1))group,with six rats in each group.The groups were given corresponding drug interventions for 21 consecutive days.The estrous cycle,the ovaries and uterus index changes were observed;the changes of fasting blood glucose(FBG)and blood lipids(triglycerides TG and total cholesterol TC)were determined by blood biochemical analyzer;fasting insulin(FINS)level was determined by enzyme-linked immunosorbent assay(ELISA);TLR4 and NF-κB p65 protein and gene expression were detected in ovarian by immunohistochemical method and real-time fluorescent quantitative PCR methods.Results Compared with the normal group,the estrous cycle in model group were disordered,the ovarian polycystic changes were obvious,the levels of FBG,TG and TC,the levels of FINS and HOMA-IR were significantly decreased,and TLR4 and NF-κB p65 proteins in ovarian tissues and mRNA expression increased(P<0.05);compared with the model group,the ovulation cycle of rats in the Cangfudaotan Decoction high-dose group was improved,the ovarian polycystic changes were reduced,and the above indicators were significant reversed(P<0.05).Conclusions Cangfudaotan Decoction could effectively improve the ovarian ovulation function in obese PCOS-IR rats and regulate glycolipid metabolism function,which may be related to the regulation of the ovarian TLR4/NF-κB p65 signaling pathway.