白芍总苷调控NF-κB/TGF-β1/Smad信号通路对慢性盆腔炎大鼠子宫组织的影响

Effects of total glucosides of paeony on uterine tissue in rats with chronic pelvic inflammatory and its mechanism

目的探讨白芍总苷(TGP)调控NF-κB/转化生长因子-β1(TGF-β1)/Smad信号通路对慢性盆腔炎大鼠子宫组织的影响。方法采用随机数字表法将120只雌性大鼠分为假手术组、模型组、妇科千金片组(150 mg/kg)和TGP低、中、高剂量组(20、40、80 mg/kg)6组,每组20只。除假手术组外,其余各组均采用机械损伤加混合细菌感染的方法制备慢性盆腔炎大鼠模型。造模10 d后,TGP低、中、高剂量组分别给予浓度4、8、16 g/L的TGP溶液(5 ml/kg)灌胃,妇科千金片组给予浓度30 g/L的妇科千金片溶液(5 ml/kg)灌胃,假手术组和模型组给予0.9%氯化钠溶液(5 ml/kg)灌胃,各组均1次/d、疗程21 d。观察子宫大体情况并通过Verco评分评价子宫粘连状况,计算子宫系数(子宫重量/大鼠体重),HE染色观察子宫组织病理学改变并行病理评分;ELISA法检测血清炎症因子TNF-α、IL-1β、IL-10水平;Western blot法检测子宫组织NF-κB、转化生长因子-β1(TGF-β1)、Smad同源物2(Smad2)、Smad同源物3(Smad3)蛋白表达水平。结果与模型组比较,妇科千金片组和TGP中、高剂量组大鼠子宫水肿状况及与周围组织粘连状况明显减轻,Verco评分和子宫系数均降低,子宫组织病变明显改善且病理评分降低,血清TNF-α、IL-1β水平均降低且IL-10水平升高,差异均有统计学意义(均P<0.05);TGP中、高剂量组子宫组织NF-κB、TGF-β1、Smad2、Smad3蛋白表达水平均下调(P<0.01)。与妇科千金片组比较,TGP高剂量组子宫系数降低,血清TNF-α水平降低、IL-10水平升高,NF-κB、TGF-β1、Smad2、Smad3蛋白表达水平均下调,差异均有统计学意义(均P<0.01)。结论TGP具有抑制慢性盆腔炎大鼠子宫粘连和组织病变的作用,其机制可能与抑制NF-κB/TGF-β1/Smad信号通路介导炎症反应有关。

Objective To investigate the effect of total glucosides of paeony(TGP)on uterine tissue in rats with chronic pelvic inflammatory and its relation to NF-κB/TGF-β1/Smad signaling pathway.Methods One hundred and twenty female SD rats were randomly divided into 6 groups:sham operation group,model group,Chinese paten medicine Fuke Qianjin tablet group(150 mg/kg)and low,medium,high dose TGP groups(20,40,80 mg/kg)with 20 rats in each group.Except for the sham operation group,the rats in other groups were given mechanical injury combined with mixed bacterial infection to induce chronic pelvic inflammatory disease.Ten days after modeling,the rats in low,medium,high dose TGP groups were given 4,8,16 g/L TGP solution(5 ml/kg),once a day for 21 d.The rats in Fuke Qianjin tablet group were given 30 g/L Fuke Qianjin tablet solution(5 ml/kg),once a day for 21 d.The rats in sham operation group and model group were given 0.9%sodium chloride solution(5 ml/kg),once a day for 21 d.The general condition of the uterus was observed and the adhesion status of the uterus was evaluated by Verco score,the uterine coefficient was calculated(uterine weight/body weight).The uterine histopathological changes was examined by HE staining and the pathological score was assessed.The content of Inflammatory factors(TNF-α,IL-1β,IL-10)in serum were detected by ELISA method.The expression of NF-κB,transforming growth factor-β1(TGF-β1)Smad2,Smad3 in uterine tissue were detected by Western blot method.Results Compared with model group,the uterine edema and adhesion to surrounding tissues of the rats in Fuke Qianjin group and medium,high dose TGP groups were attenuated,and the Verco score and the uterine coefficient were reduced,the uterine tissue lesions were significantly improved and the pathological scores were reduced,the level of TNF-α,IL-1β in serum were decreased while IL-10 level was increased(all P<0.05).The expression of NF-κB,TGF-β1,Smad2,Smad3 in uterine tissue of medium,high dose TGP groups were down-regulated(all P<0.01).Compared with Fuke Qianjin group,the uterine coefficient in high dose TGP group was reduced,the level of TNF-αin serum were decreased and the level of IL-10 was increased,the expressions of NF-κB,TGF-β1,Smad2,Smad3 were down-regulated(all P<0.01).Conclusion TGP has the effect of inhibiting uterine adhesion and attenuating tissue pathology in rats with chronic pelvic inflammatory disease,which may be related to the inhibition of NF-κB/TGF-β1/Smad signaling pathway to mediate inflammation.