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

作　　者：谭炜富 晏丽君 杨丽玲 余景滔[2] 卢子滨 周湘君 杨广丽 李薇 余林中 TAN Weifu;YAN Lijun;YANG Liling;YU Jingtao;LU Zibin;ZHOU Xiangjun;YANG Guangli;LI Wei;YU Linzhong(Dongguan Hospital Affiliated to Jinan University,Dongguan 523900,China;Pharmacology Laboratory of Chinese Medicine,State Administration of Traditional Chinese Medicine Research Level III Laboratory,School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China;Guangdong Key Laboratory of Natural Drug Research and Development,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]暨南大学附属东莞医院,广东东莞523900 [2]南方医科大学中医药学院国家中医药管理局科研三级实验室中药药理实验室,广东广州510515 [3]广东医科大学广东天然药物研究与开发重点实验室,广东东莞523808

出　　处：《中国中医药信息杂志》2023年第8期29-35,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：东莞市社会发展科技项目(20211800904592、2022800905232);国家自然科学基金(82204718、82074317、81730110);广东省基础与应用基础研究基金项目(2019A1515110369);广东省中医药局科研项目(20211411)。

摘　　要：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.

关 键 词：凉膈散 内毒素 急性肺损伤 GSK-3Β Th17/Treg比值 

分 类 号：R259.63[医药卫生—中西医结合] R285[医药卫生—中医内科学]