自拟补肾祛瘀方对多囊卵巢综合征模型大鼠的作用及机制研究

Efficacy and mechanism analysis of self-made Bushen Quyu decoction in treatment of polycystic ovary syndrome model rats

目的探究自拟补肾祛瘀方治疗多囊卵巢综合征(PCOS)的疗效及机制。方法选取SD雌性大鼠,复制PCOS模型后随机分为对照组、模型组、西药组及联合组。对照组和模型组根据大鼠体重给予1 mL/(kg·d)氯化钠溶液灌胃,联合组每天根据体重给予补肾祛瘀汤剂11 g/kg和4.5 mg/kg二甲双胍灌胃,西药组仅给予4.5 mg/kg二甲双胍灌胃。药物干预后分别采用Western blotting、qRT-PCR、免疫组织化学法检测肿瘤坏死因子-α(TNF-α)、核因子κB(NF-κB)、核因子IκB(IκBα)的蛋白、mRNA及阳性表达。比较各组大鼠的体重及血清睾酮(T)、促卵泡激素(FSH)、黄体生成素(LH)水平。结果各组大鼠药物干预前的体重比较,差异无统计学意义(P>0.05);各组大鼠药物干预28 d后的体重比较,差异有统计学意义(P<0.05);进一步两两比较,模型组的体重高于另外3组(P<0.05),西药组高于联合组和对照组(P<0.05),联合组高于对照组(P<0.05)。各组大鼠血清T、FSH、LH水平比较,差异有统计学意义(P<0.05);进一步两两比较,模型组血清T、FSH、LH水平高于另外3组(P<0.05),西药组高于联合组与对照组(P<0.05),联合组高于对照组(P<0.05)。各组大鼠卵巢组织TNF-α、NF-κB、IκBα蛋白相对表达量比较,差异有统计学意义(P<0.05);进一步两两比较,模型组大鼠卵巢组织TNF-α、NF-κB蛋白相对表达量高于另外3组,IκBα蛋白相对表达量低于另外3组(P<0.05);西药组大鼠卵巢组织TNF-α、NF-κB蛋白相对表达量高于联合组与对照组(P<0.05),联合组高于对照组(P<0.05);西药组大鼠卵巢组织IκBα蛋白相对表达量低于联合组与对照组(P<0.05),联合组低于对照组(P<0.05)。各组大鼠卵巢组织TNF-α、NF-κB、IκBαmRNA相对表达量比较,差异有统计学意义(P<0.05);进一步两两比较,模型组大鼠卵巢组织TNF-α、NF-κB mRNA相对表达量高于另外3组,IκBαmRNA相对表达量低于另外3组(P<0.05);西药组大鼠卵巢组织TNF-α、NF-κB mRNA相对表达量高于联合组与对照组(P<0.05),联合组高于对照组(P<0.05);西药组大鼠卵巢组织IκBαmRNA相对表达量低于联合组与对照组(P<0.05),联合组低于对照组(P<0.05)。各组大鼠卵巢组织TNF-α、NF-κB、IκBα的OD值比较,差异有统计学意义(P<0.05);进一步两两比较,模型组大鼠卵巢组织TNF-α、NF-κB的OD值高于另外3组,IκBα的OD值低于另外3组(P<0.05);西药组大鼠卵巢组织TNF-α、NF-κB的OD值高于联合组与对照组(P<0.05),联合组高于对照组(P<0.05);西药组大鼠卵巢组织IκBα的OD值低于联合组与对照组(P<0.05),联合组低于对照组(P<0.05)。结论自拟补肾化瘀方治疗PCOS可通过影响TNF-α调控的NF-κB/IκBα信号通路促进LH、FSH、T的恢复。

Objective To explore the effect and mechanism of self-made Bushen Quyu Decoction in treatment of polycystic ovary syndrome(PCOS).Methods Six-week-old SPF SD female rats were selected and randomly divided into control group,model group,western medicine group,and combination group after letrozole modeling.After drug intervention,Western blotting,RT-PCR,and immunohistochemistry were used to detect the expression of tumor necrosis factor-α(TNF-α),nuclear factorκB(NF-κB),and nuclear factor IκB(IκBα).The body weight and serum testosterone(T),follicle-stimulating hormone(FSH),and luteinizing hormone(LH)were compared in each group before and after treatment.Results Before treatment,the differences in body weight of the rats in each group were tested by one-way ANOVA,and the difference was not statistically significant(PP>0.05).After treatment,the differences in the body weight of the rats in each group were statistically significant by one-way ANOVA test(P<0.05).Further pairwise comparisons showed that the body weight of the model group after treatment was higher than that of the other three groups(P<0.05),the body weight of the group was higher than that of the combined group and the control group(P<0.05),and the body weight of the combined group was higher than that of the control group(P<0.05).The differences between groups in abdominal aortic blood T,FSH and LH of rats in each group were statistically significant(P<0.05).Further pairwise comparison showed that T,FSH and LH in the model group were higher than those in the other three groups(P<0.05);at the same time,the western medicine group was higher than the combination group and the control group(P<0.05),and the combination group was higher than the control group(P<0.05).The differences in the protein expression,mRNA expression,and OD value of TNF-α,NF-κB,and IκBαin ovarian tissue of each group were statistically significant by one-way ANOVA test(P<0.05).Further pairwise comparison showed that the expression of TNF-α,NF-κB protein,mRNA expression,and OD value in the ovarian tissue of the model group were higher than those of the other three groups,while the protein expression,m RNA expression and OD value of IκBαwere lower than those of the other three groups(P<0.05);TNF-α,NF-κB protein expression,m RNA expression,and OD value in the ovarian tissue of the western medicine group were higher than those of the combination group and the control group(P<0.05),and these of the combination group was higher than those of the control group(P<0.05);the expression of IκBαprotein,mRNA expression,and OD in ovarian tissue of the western medicine group were lower than those of the combination group and the control group(P<0.05),and these of the combination group was lower than those of the control group(P<0.05).Conclusion The self-made Bushen Huayu Decoction for PCOS can promote the recovery of LH,FSH and T by regulating TNF-α/NF-κB/IκBαsignaling pathway.