基于网络药理学和动物实验探讨乌药治疗慢性盆腔炎的作用机制

Mechanisms of Wuyao(Linderae Radix)in treating chronic pelvic inflammatory disease based on network pharmacology and animal experiments

目的基于网络药理学预测乌药治疗慢性盆腔炎(CPID)的活性成分和作用靶点,通过动物实验验证并探讨其可能的作用机制,为乌药的临床应用提供科学依据。方法运用网络药理学和分子对接技术筛选乌药治疗CPID的可能有效成分及其作用靶点;采用大肠杆菌,金黄色葡萄球菌和解脲脲原体的混合菌加机械损伤法建立慢性盆腔炎模型大鼠;采用HE染色观察子宫、输卵管及脾脏的病理形态变化;采用试剂盒法检测大鼠血清中一氧化氮(NO)、超氧化物歧化酶(SOD)、丙二醛(MDA)含量;酶联免疫吸附试验(ELISA)检测大鼠血清中白介素(IL)-6、IL-10的表达水平。流式细胞术法检测脾脏CD4^(+)、CD8a^(+)T细胞及CD4^(+)CD25^(+)调节性T细胞(Tregs)数量百分比。结果网络药理学研究共筛选出乌药中9个有效活性成分和4个核心治疗靶点,它们主要影响炎症反应相关的免疫调节。动物研究表明,乌药可显著抑制慢性盆腔炎大鼠子宫肿胀,调节输卵管及脾脏的形态改变,并有效减轻子宫和输卵管的炎性浸润与损伤,改善脾脏功能;乌药可显著降低血清中NO、IL-6和MDA水平,升高IL-10和SOD水平,并提高模型大鼠CD4^(+)T细胞数量百分比、升高CD4^(+)T/CD8a^(+)T细胞比值,降低CD8a^(+)T细胞数量百分比,提高模型大鼠异常降低的CD4^(+)CD25^(+)Tregs数量百分比(P<0.05)。结论乌药通过减轻慢性盆腔炎模型大鼠的氧化应激和炎症水平,同时调节T细胞亚群的免疫功能,从而改善慢性盆腔炎的病理过程而达到治疗效果,其临床价值值得进一步探索。

Objective To predict the active components and action targets of Wuyao(Linderae Radix)in the treatment of chronic pelvic inflammatory disease(CPID)based on network pharmacology,explore possible mechanisms of the treatment through animal experiments,and provide a scientific basis for clinical applications of Wuyao(Linderae Radix).Methods Possible active components and targets of Wuyao(Linderae Radix)in the treatment of CPID were obtained applying network pharmacology and molecular docking technology.CPID rat models were established using the mixed Escherichia coli,Staphylococcus aureus,and Ureaplasma urealyticum plus the performance of mechanical injury.Hematoxylineosin(HE)staining was applied to observe the pathological changes in the uterus,fallopian tube,and spleens of rat models.The contents of nitric oxide(NO),superoxide dismutase(SOD),and malondialdehyde(MDA)in the serum of rats were determined with the use of corresponding detection kits.Enzyme-linked immunosorbent assay(ELISA)test was used to measure the expression of interleukin(IL)-6 and IL-10 in the serum of rat models.Flow cytometry was used to determine the percentage of CD4^(+)and CD8a^(+)T cells as well as CD4^(+)CD25^(+)regulatory T cells(Tregs)in the spleen of rat models.Results A total of nine potential active components and four core therapeutic targets related to inflammatory response in Wuyao(Linderae Radix)were obtained.The animal experiments showed that Wuyao(Linderae Radix)markedly inhibited uterus swelling,regulated morphological changes in the fallopian tube and spleen,effectively reduced inflammatory infiltration and injuries in the uterus and fallopian tube,and improved spleen functions in CPID rats.Moreover,Wuyao(Linderae Radix)markedly reduced the levels of NO,IL-6,and MDA,and increased the levels of IL-10 and SOD in the serum of rats.Wuyao(Linderae Radix)also elevated the percentage of CD4^(+)T cells and the CD4^(+)T/CD8a^(+)T cell ratio,reduced the percentage of CD8a^(+)T cells,and raised the percentage of CD4^(+)CD25^(+)Tregs that had been abnormally decreased in rat models(P<0.05).Conclusion Wuyao(Linderae Radix)could have therapeutic effects on CPID rats by relieving oxidative stress,mitigating inflammatory levels,and regulating the immuno-function of T cell subgroups to improve the pathological changes in CPID rats.It is a medicinal herb worth being further explored for its clinical values.