苓甘五味姜辛汤治疗急性肺损伤的作用机制

Study on Mechanism of Linggan Wuwei Jiangxintang in Treating Acute Lung Injury

目的:探究苓甘五味姜辛汤治疗脂多糖(LPS)诱导的小鼠急性肺损伤(ALI)的作用机制。方法:使用中药系统药理学数据库与分析平台(TCMSP)数据库和GeneCards、DisGeNET、Herb数据库结合基因表达综合数据库(GEO)临床数据筛选苓甘五味姜辛汤治疗ALI的关键靶点,构建蛋白质-蛋白质相互作用网络(PPI)筛选核心靶点,并进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析。使用LPS诱导建立小鼠ALI模型,验证苓甘五味姜辛汤治疗ALI的效果及相关靶点,并采用蛋白免疫印迹法(Western blot)检测肺组织中Toll样受体(TLR)4、核转录因子-κB p65(NF-κB p65)、磷酸化NF-κB p65(p-NF-κB p65)的表达水平。结果:分析显示苓甘五味姜辛汤治疗ALI与白细胞介素-6(IL-6)、肿瘤坏死因子(TNF)-α、原癌基因(JUN)等10个核心靶点相关,涉及TNF信号通路、TLR信号通路、NF-κB信号通路等。动物实验结果显示苓甘五味姜辛汤可以减轻肺部损伤,改善ALI小鼠的病理状态,显著降低血清中TNF-α、IL-6的表达,提高肺组织中总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)的活性,显著降低肺组织中JUN、TLR4、NF-κB p65、NF-κB p-p65蛋白表达水平。结论:苓甘五味姜辛汤可以抑制LPS诱导的ALI小鼠炎症和氧化损伤,其机制可能与抑制TLR4/NF-κB信号通路,降低炎症因子TNF-α、IL-6的水平有关。

Objective:To explore the action mechanism of Linggan Wuwei Jiangxintang on the treatment of lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice.Method:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),GeneCards,DisGeNET,and Herb databases were combined with clinical data from Gene Expression Omnibus(GEO)to screen the key targets of Linggan Wuwei Jiangxintang in the treatment of ALI.The protein-protein interaction(PPI)network was constructed to screen the core targets,and gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analyses were performed.The mouse ALI model was established by LPS induction to verify the effect and key targets of Linggan Wuwei Jiangxintang on the treatment of ALI.The expression levels of Toll-like receptor 4(TLR4),nuclear transcription factor-κB p65(NF-κB p65),and phosphorylated NF-κB p65(NF-κB p-p65)in lung tissue were detected by Western blot.Result:The analysis showed that the treatment of ALI with Linggan Wuwei Jiangxintang was related to 10 core targets such as interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and JUN,involving TNF signaling pathway,Toll-like receptor signaling pathway,NF-κB signaling pathway,etc.The animal experimental results show that Linggan Wuwei Jiangxintang can reduce lung injury,improve the pathological state of ALI mice,significantly reduce the expression of TNF-αand IL-6 in serum,increase the activity of total superoxide dismutase(T-SOD)and catalase(CAT)in lung tissue,and reduce the expression levels of JUN,TLR4,NF-κB p65,and NF-κB p-p65 proteins in lung tissue.Conclusion:Linggan Wuwei Jiangxintang can inhibit LPS-induced inflammation and oxidative damage in ALI mice,and its mechanism may be related to the inhibition of TLR4/NF-κB signaling pathway and the reduction of inflammatory factors such as TNF-αand IL-6.