经Toll样受体4/核因子κB通路探讨复方黄柏液涂剂对细菌性阴道病模型大鼠Th1/Th2的调节及抑炎机制

Roles of compound cortex phellodendri fluid in anti⁃inflammatory and regulating Th1/Th2 of bacterial vaginosis model rats via TLR4/NF⁃κB pathway

目的:通过Toll样受体(Toll⁃like receptors,TLR)4/核因子κB(NF⁃κB)通路探索复方黄柏液涂剂治疗细菌性阴道病模型大鼠的疗效及作用机制。方法:选取55只SPF级SD雌性大鼠,采用混合菌液接种法制备细菌性阴道病大鼠模型。造模成功后分别给予复方黄柏液涂剂、甲硝唑阴道凝胶、红核妇洁洗液连续治疗7 d,HE染色观察大鼠阴道组织中炎性细胞浸润及糖原堆积情况;酶联免疫吸附法(ELISA)检测大鼠阴道灌洗液中白介素(IL)⁃1β,IL⁃2,IL⁃10,IL⁃6和肿瘤坏死因子(TNF)⁃α表达水平;免疫组化定位大鼠阴道组织中NF⁃κB p65分布情况;Western blot法检测大鼠阴道组织中TLR4和NF⁃κB p65蛋白表达水平。结果:与模型组相比,黄柏组大鼠阴道组织中炎性细胞浸润减轻,糖原堆积明显增多(P<0.01),阴道灌洗液中IL⁃1β,IL⁃6浓度降低(P<0.01,P<0.05),IL⁃2,TNF⁃α浓度下降,IL⁃10浓度升高(P>0.05),TLR4,NF⁃κB p65蛋白表达水平均下降(P<0.05),NF⁃κB p65核易位细胞数减少(P<0.01)。结论:复方黄柏液涂剂对细菌性阴道病模型大鼠具有治疗作用,其机制可能与抑制TLR4/NF⁃κB通路、减少炎症介质分泌、调节Th1/Th2免疫失衡有关。

Objective:To explore the effect and mechanism of compound cortex phellodendri fluid in bacterial vaginosis rats via TLR4/NF⁃κB pathway.Methods:Fifty⁃five SD female rats were selected to construct bacterial vaginosis model by injection of mixed bacterial liquid.After successful modeling,the rats were treated with compound cortex phellodendri fluid,metronidazole vaginal gel,and Honghe fujie lotion,respectively.After 7 d of continuous treatment,inflammatory cell infiltration and glycogen accumulation in rat vaginal tissues were observed by HE staining.ELISA method was used to detect the expression level of IL⁃1β,IL⁃2,IL⁃10,IL⁃6,and TNF⁃αin the vaginal lavage.The distribution of NF⁃κB p65 in rat vaginal tissues were located by immunohistochemical method.And the protein expression level of TLR4 and NF⁃κB p65 were detected by Western blot.Results:Compared with those of the model group,the infiltration of inflammatory cells in compound cortex phellodendri fluid group was reduced,and the accumulation of glycogen was significantly increased(P<0.01),the concentrations of IL⁃1β(P<0.01),IL⁃6(P<0.05),IL⁃2,and TNF⁃αin the vaginal lavage were decreased,while the IL⁃10 concentration increased(P>0.05),the protein expression level of TLR4 and NF⁃κB p65 decreased(P<0.05),and the NF⁃κB p65's translocated nuclear were suppressed(P<0.01).Conclusion:Compound cortex phellodendri fluid has a definite therapeutic effect on bacterial vaginosis rats.Its mechanism may be related to the inhibition of TLR4/NF⁃κB pathway and adjustion of immune imbalance on Th1/Th2.