摘要
目的 通过网络药理学结合体外细胞实验探究葛根芩连汤治疗肾纤维化的潜在作用。方法 借助本草组鉴(HERB)、 PubChem数据库、 SwissADME平台及SwissTargetPrediction数据库获取葛根芩连汤的主要活性成分及作用靶点,利用DisGenet数据库获得肾脏纤维化相关的靶点基因。同时通过蛋白质免疫印迹法(WB)检测TGF-β1表达。利用RT-qPCR法检测Bcl-2、 STAT3和FN1表达。结果 筛选并得到葛根芩连汤与肾纤维化的共同作用靶点157个。GO/KEGG富集分析筛选GeneRatio在前15名的细胞成分、生物过程、分子功能和信号通路,这些靶点主要参与各种氧化应激反应,并通过MAPK信号通路和P13K-Akt信号通路调控细胞增殖和凋亡。体外实验证明葛根芩连汤含药血清干预可降低TGF-β1诱导HK-2细胞TGF-β1、 STAT3和FN1的表达,同时提高Bcl-2的表达水平。结论 葛根芩连汤改善肾脏纤维化可能与抑制TGF-β的表达以减少EMT发生,同时通过上调凋亡相关基因Bcl-2的表达、抑制STAT3的表达从而阻断细胞凋亡以减轻或逆转炎症状态有关。
Objective To explore the potential effects of Gegen Qinlian decoction(GGQLD)on renal fibrosis based on network pharmacology combined with in vitro cell experiments.Methods The main active ingredients and targets of GGQLD were identified by means of the Herbal Biochemistry(HERB),PubChem database,SwissADME platform,and SwissTargetPrediction database.The target genes related to renal fibrosis were obtained from the DisGenet database.Western blotting(WB)assay was performed to detect the expression of TGF-β1,and RT-qPCR assay was used to detect the expression of Bcl-2,STAT3 and FN1.Results 157 common targets of GGQLD and renal fibrosis were screened and identified.GO/KEGG enrichment analysis was performed to identify the cell components,biological processes,molecular functions and signaling pathways of the top items in GeneRatio.It was found that these targets were mainly involved in various oxidative stress responses and regulated cell proliferation and apoptosis through the MAPK signaling pathway and P13K Akt signaling pathway.In vitro experiments showed that the intervention of serum containing GGQLD could reduce the expression of TGF-β1,STAT3 and FN1 induced by TGF-β1 in HK-2 cells,while increased the expression of Bcl-2.Conclusion The improvement of renal fibrosis by GGQLD may be related to the inhibition of TGF-βexpression to reduce the occurrence of epithelial-mesenchymal transition(EMT).At the same time,it is also related to its ability to block cell apoptosis by up-regulating the expression of apoptosis related gene Bcl-2 and inhibit the expression of STAT3,thereby reducing or reversing the inflammatory state.
作者
刘德华
王骁珺
陆黎黎
顾徐超
孙怡婕
Liu Dehua;Wang Xiaojun;Lu Lili;Gu Xuchao;Sun Yijie(Department of Traditional Chinese Medicine,Huadong Hospital Affiliated to Fudan University,Shanghai,200040,P.R.China;Clinical Key Laboratory of Geriatric Medicine of Shanghai City,Shanghai,200040,P.R.China)
出处
《老年医学与保健》
CAS
2024年第5期1419-1424,共6页
Geriatrics & Health Care
基金
国家自然科学基金(82474417)。
