基于网络药理学探讨环黄芪醇治疗肾纤维化的作用机制

Study on the Mechanism of Cycloastragenol in Treating Renal Fibrosis Based on Network Pharmacology

目的探讨环黄芪醇治疗肾纤维化的作用靶点和机制。方法采用网络药理学方法预测环黄芪醇标靶肾纤维化的核心靶点,并对其进行GO富集分析和KEGG通路分析。使用分子对接技术验证环黄芪醇与核心靶点的结合性能。构建单侧输尿管结扎的小鼠肾纤维化模型,连续灌胃环黄芪醇,分别于7 d、14 d和21 d后终止。检测肾体比、BUN和CRE水平。采用HE和Masson染色,观察肾脏组织病理学变化。qRT-PCR和Western Blotting测定肾脏组织中TGF-β1、α-SMA、MMP3和MMP2的表达水平。结果网络药理学和分子对接结果显示,环黄芪醇标靶肾纤维化成功对接8个靶点,其中,MMP3和MMP2的结合能最佳。动物实验结果显示,环黄芪醇能有效改善肾纤维化小鼠肾脏胶原纤维沉积,降低TGF-β1、α-SMA、MMP3和MMP2的表达(P<0.05)。随着环黄芪醇剂量增大和用药时间增加,各指标的阳性表达呈减弱趋势。结论环黄芪醇可能是通过作用于MMP3和MMP2等靶点,调解Rap1信号通路(Rap1 signaling pathway)、IL-17信号通路(IL-17 signaling pathway)、黄体酮介导的卵母细胞成熟(Progesterone-mediated oocyte maturation)和脂质与动脉粥样硬化(Lipid and atherosclerosis)等通路作用,发挥治疗肾纤维化的作用。

Objective To investigate the target and mechanism of cycloastragenol in treating renal fibrosis.Methods The core target of cycloastragenol targeted renal fibrosis was predicted by network pharmacology,and its GO enrichment and KEGG pathway were analyzed.Molecular docking technology was used to verify the binding properties of cycloastragenol to the core target.A mouse model of renal fibrosis with unilateral ureteral ligation was established,and continuous intragastacal administration was terminated after 7 d,14 d and 21 d,respectively.The renal body ratio,BUN and CRE levels were detected.The histopathological changes of kidney were observed by HE and Masson staining.The expression levels of TGF-β1,α-SMA,MMP3 and MMP2 in renal tissues were determined by qRT-PCR and Western Blotting.Results Network pharmacology and molecular docking results showed that 8 targets of renal fibrosis were successfully docked with cycloastragenol,among which MMP3 and MMP2 had the best binding energy.The results of animal experiments showed that cycloastragenol could effectively improve the deposition of renal collagen fiber and reduce the expression of TGF-β1,α-SMA,MMP3 and MMP2 in mice with renal fibrosis,with statistical significance(P<0.05).With the increase of dosage and time of administration,the positive expression of each index showed a decreasing trend.Conclusion Cycloastragenol may act on targets such as MMP3 and MMP2,mediated Rap1 signaling pathway,IL-17 signaling pathway,and Progesteronemediated oocyte maturation,Lipid and atherosclerosis pathways play a role in the treatment of renal fibrosis.