基于生物信息学与实验验证探讨凉膈散通过GSK-3β调控内毒素致急性肺损伤分子机制

Exploration on the Molecular Mechanism of Endotoxin-induced Acute Lung Injury Through GSK-3βRegulation by Liangge Powder Based on Bioinformatics and Experimental Validation

目的探索凉膈散治疗内毒素所致急性肺损伤(ALI)的作用靶点,分析并验证其可能的作用机制。方法通过GEO、TCMSP和中医药整合药理学研究平台(TCMIP)获取凉膈散PI3K/Akt通路相关ALI疾病基因,采用单基因Meta分析识别凉膈散可能的作用靶点,通过脂多糖诱导的小鼠ALI模型对预测结果进行验证。结果生物信息学分析确定GSK3β、CDKN1A、NFKB1和MCL1为PI3K/Akt通路相关的ALI疾病特征基因,其中GSK3β是凉膈散治疗ALI的作用靶点。在脂多糖诱导的ALI小鼠模型中,凉膈散和GSK-3β抑制剂TWS119均可显著减轻小鼠肺组织水肿(P<0.001),下调GSK-3β基因表达,降低Th17细胞比例、Th17/Treg比值和白细胞介素-17含量(P<0.001,P<0.05),提高Treg细胞比例和转化生长因子-β含量(P<0.05)。加入GSK-3β激动剂Wortmannin处理后,凉膈散的上述作用被部分逆转。结论GSK-3β是凉膈散治疗ALI的作用靶点,凉膈散可通过抑制GSK-3β表达调节Th17/Treg免疫平衡,从而发挥治疗ALI作用。

Objective To explore the targets of Liangge Powder in the treatment of endotoxin-induced acute lung injury(ALI);To analyze and verify its possible mechanisms of action.Methods The Liangge Powder-PI3K/Akt pathway-related ALI disease genes were obtained through public databases such as GEO,TCMSP and TCMIP.Singlegene meta-analysis was used to identify possible targets of Liangge Powder.The predicted results were validated using a mouse ALI model induced by lipopolysaccharide.Results Bioinformatics analysis identified GSK3β,CDKN1A,NFKB1 and MCL1 as PI3K/Akt pathway-related ALI disease signature genes,of which GSK3βwas the target of Liangge Powder for ALI treatment.In the lipopolysaccharide induced ALI mouse model,Liangge Powder and GSK-3βinhibitor TWS119 could significantly reduce lung tissue edema in mice(P<0.001),lower GSK-3βgene expression,reduce the proportion of Th17 cells,Th17/Treg ratio,and interleukin-17 content(P<0.001,P<0.05),and increase the proportion of Treg cells and transform growth factor-βcontent(P<0.05).These effects of Liangge Powder were partially reversed by the addition of the GSK-3βagonist Wortmannin treatment.Conclusion GSK-3βis the target of Liangge Powder for the treatment of ALI,and Liangge Powder can treat ALI by regulating the Th17/Treg immune balance by inhibiting GSK-3βexpression.