Objective The main pathological feature of immunoglobulin A nephropathy(IgAN),an autoimmune kidney disease,is the deposition of IgA immune complexes,accompanied by mesangial cell proliferation and elevated urine prote...Objective The main pathological feature of immunoglobulin A nephropathy(IgAN),an autoimmune kidney disease,is the deposition of IgA immune complexes,accompanied by mesangial cell proliferation and elevated urine protein.The Guben Tongluo formula(GTF)is a traditional Chinese medicine prescription,which has predominant protective effects on IgAN.However,the therapeutic mechanism of the GTF in IgAN remains elusive.The present study aimed to determine the effects of GTF in treating IgAN via regulating the TLR4/MyD88/NF-κB pathway.Methods In the present study,lamina propria B lymphocytes were treated with different concentrations of lipopolysaccharide(LPS)(0,1,5,10 and 20 ng/mL).Flow cytometry was used to define positive CD86+CD19+cells.CCK-8 assay was used to examine cell proliferation.RNAi was used to induce TLR4 silencing.qRT-PCR and Western blotting were used to determine gene expression.Results It was found that the LPS dose-dependently increased the content of IgA and galactose-deficient IgA1(Gd-IgA),the levels of TLR4,Cosmc,MyD88 and phosphorylated(p)-NF-κB,and the ratio of CD86+CD19+and IgA-producing B cells.However,the TLR4 knockdown reversed the role of LPS.This suggests that TLR4 mediates the effects of LPS on lamina propria B lymphocytes.Furthermore,the GTF could dose-dependently counteract the effects of LPS and TLR4 overexpression on lamina propria B lymphocytes through the TLR4/MyD88/NF-κB pathway.Conclusion Collectively,these results demonstrate that the GTF can regulate the TLR4/MyD88/NF-κB pathway to treat IgAN model lamina propria B lymphocytes stimulated by LPS.展开更多
目的运用质谱分析、网络药理学和动物实验探讨健脑固本方治疗脑小血管病性认知障碍(cerebral small vessel disease,CSVCI)的作用机制。方法通过质谱分析得出主要活性成分,TCMSP检索其相应作用靶点;通过GeneCards、OMIM数据库检索CSVCI...目的运用质谱分析、网络药理学和动物实验探讨健脑固本方治疗脑小血管病性认知障碍(cerebral small vessel disease,CSVCI)的作用机制。方法通过质谱分析得出主要活性成分,TCMSP检索其相应作用靶点;通过GeneCards、OMIM数据库检索CSVCI疾病靶点;通过String数据库和Cytoscape软件构建药物疾病作用靶点网络可视化;利用DAVID数据库对交集靶点进行GO功能和KEGG信号通路富集分析。结果健脑固本方的有效活性成分有山奈酚、薯蓣皂苷,作用于丝裂原活化蛋白激酶8、蛋白激酶B1等核心靶点,可能通过磷脂酰肌醇3-激酶/蛋白激酶B等信号通路涉及炎症反应。蛋白免疫印迹法检测结果表明健脑固本方抑制Notch-1、RBP-Jκ以及Hes-1的表达。结论本研究初步表明健脑固本方能够通过多靶点、多通路、多途径治疗CSVCI,具体作用机制可能是抑制Notch-1信号通路,进而抑制炎症反应。展开更多
基金supported by the National Natural Science Foundation of China(No.81904124)the Scientific Project of Shanghai Sanitation and Health Committee(No.20204Y0191)the projects of Shanghai Science and Technology Commission(No.22Y31920200).
文摘Objective The main pathological feature of immunoglobulin A nephropathy(IgAN),an autoimmune kidney disease,is the deposition of IgA immune complexes,accompanied by mesangial cell proliferation and elevated urine protein.The Guben Tongluo formula(GTF)is a traditional Chinese medicine prescription,which has predominant protective effects on IgAN.However,the therapeutic mechanism of the GTF in IgAN remains elusive.The present study aimed to determine the effects of GTF in treating IgAN via regulating the TLR4/MyD88/NF-κB pathway.Methods In the present study,lamina propria B lymphocytes were treated with different concentrations of lipopolysaccharide(LPS)(0,1,5,10 and 20 ng/mL).Flow cytometry was used to define positive CD86+CD19+cells.CCK-8 assay was used to examine cell proliferation.RNAi was used to induce TLR4 silencing.qRT-PCR and Western blotting were used to determine gene expression.Results It was found that the LPS dose-dependently increased the content of IgA and galactose-deficient IgA1(Gd-IgA),the levels of TLR4,Cosmc,MyD88 and phosphorylated(p)-NF-κB,and the ratio of CD86+CD19+and IgA-producing B cells.However,the TLR4 knockdown reversed the role of LPS.This suggests that TLR4 mediates the effects of LPS on lamina propria B lymphocytes.Furthermore,the GTF could dose-dependently counteract the effects of LPS and TLR4 overexpression on lamina propria B lymphocytes through the TLR4/MyD88/NF-κB pathway.Conclusion Collectively,these results demonstrate that the GTF can regulate the TLR4/MyD88/NF-κB pathway to treat IgAN model lamina propria B lymphocytes stimulated by LPS.
文摘目的运用质谱分析、网络药理学和动物实验探讨健脑固本方治疗脑小血管病性认知障碍(cerebral small vessel disease,CSVCI)的作用机制。方法通过质谱分析得出主要活性成分,TCMSP检索其相应作用靶点;通过GeneCards、OMIM数据库检索CSVCI疾病靶点;通过String数据库和Cytoscape软件构建药物疾病作用靶点网络可视化;利用DAVID数据库对交集靶点进行GO功能和KEGG信号通路富集分析。结果健脑固本方的有效活性成分有山奈酚、薯蓣皂苷,作用于丝裂原活化蛋白激酶8、蛋白激酶B1等核心靶点,可能通过磷脂酰肌醇3-激酶/蛋白激酶B等信号通路涉及炎症反应。蛋白免疫印迹法检测结果表明健脑固本方抑制Notch-1、RBP-Jκ以及Hes-1的表达。结论本研究初步表明健脑固本方能够通过多靶点、多通路、多途径治疗CSVCI,具体作用机制可能是抑制Notch-1信号通路,进而抑制炎症反应。
