A systematic study of Erzhu Erchen decoction against damp-heat internalized type 2 diabetes based on data mining and experimental verification

Background:Erzhu Erchen decoction(EZECD),which is based on Erchen decoction and enhanced with Atractylodes lancea and Atractylodes macrocephala,is widely used for the treatment of dampness and heat(The clinical manifestations of Western medicine include thirst,inability to drink more,diarrhea,yellow urine,red tongue,et al.)internalized disease.Nevertheless,the mechanism of EZECD on damp-heat internalized Type 2 diabetes(T2D)remains unknown.We employed data mining,pharmacology databases and experimental verification to study how EZECD treats damp-heat internalized T2D.Methods:The main compounds or genes of EZECD and damp-heat internalized T2D were obtained from the pharmacology databases.Succeeding,the overlapped targets of EZECD and damp-heat internalized T2D were performed by the Gene Ontology,kyoto encyclopedia of genes and genomes analysis.And the compound-disease targets-pathway network were constructed to obtain the hub compound.Moreover,the hub genes and core related pathways were mined with weighted gene co-expression network analysis based on Gene Expression Omnibus database,the capability of hub compound and genes was valid in AutoDock 1.5.7.Furthermore,and violin plot and gene set enrichment analysis were performed to explore the role of hub genes in damp-heat internalized T2D.Finally,the interactions of hub compound and genes were explored using Comparative Toxicogenomics Database and quantitative polymerase chain reaction.Results:First,herb-compounds-genes-disease network illustrated that the hub compound of EZECD for damp-heat internalized T2D could be quercetin.Consistently,the hub genes were CASP8,CCL2,and AHR according to weighted gene co-expression network analysis.Molecular docking showed that quercetin could bind with the hub genes.Further,gene set enrichment analysis and Gene Ontology represented that CASP8,or CCL2,is negatively involved in insulin secretion response to the TNF or lipopolysaccharide process,and AHR or CCL2 positively regulated lipid and atherosclerosis,and/or including NOD-like receptor signaling pathway,and TNF signaling pathway.Ultimately,the quantitative polymerase chain reaction and western blotting analysis showed that quercetin could down-regulated the mRNA and protein experssion of CASP8,CCL2,and AHR.It was consistent with the results in Comparative Toxicogenomics Database databases.Conclusion:These results demonstrated quercetin could inhibit the expression of CASP8,CCL2,AHR in damp-heat internalized T2D,which improves insulin secretion and inhibits lipid and atherosclerosis,as well as/or including NOD-like receptor signaling pathway,and TNF signaling pathway,suggesting that EZECD may be more effective to treat damp-heat internalized T2D.

二陈汤与苓桂术甘汤治疗非酒精性脂肪性肝病炎性损伤的机制研究

[目的]探讨经验方二陈汤与苓桂术甘汤对高脂饲料诱导大鼠非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)分子作用机制。[方法]以高脂饲料喂养40只雄性SD大鼠8周,随机分为正常组、模型组、罗格列酮组、二陈汤组、苓桂术甘汤组,于第9周起分别给予罗格列酮剂量1mg·kg-1、二陈汤4.52g·kg-1、苓桂术甘汤组3.31g·kg-1治疗4周。12周后将大鼠处死并观察药物对大鼠肝功能的影响,并对肝组织进行常规苏木精-伊红染色,观察病理改变;Western blot检测二陈汤与苓桂术甘汤以及两方含药血清对NASH大鼠肝组织以及经软脂酸刺激肝细胞的肿瘤坏死因子TNF-α与核转录因子NF-κB蛋白表达的影响。MTT法检测不同浓度软脂酸对大鼠肝细胞增殖的影响,以及二陈汤与苓桂术甘汤含药血清对软脂酸的拮抗作用。[结果]与模型组比较,罗格列酮和苓桂术甘汤可有效降低NASH大鼠血清ALT、AST(P<0.05),同时改善肝脂肪变性的严重程度和降低TNF-α和NF-κB蛋白的表达(P<0.05)。与模型组比较,虽然二陈汤治疗后NASH大鼠肝组织TNF-α和NF-κB蛋白表达显著性下降(P<0.05),但是血清ALT、AST并未显著性降低(P>0.05),苓桂术甘汤含药血清能明显改善软脂酸对肝细胞增殖的抑制(P<0.05)。与模型组比较,二陈汤与苓桂术甘汤含药血清均能抑制软脂酸刺激的肝细胞TNF-α和NF-κB蛋白表达(P<0.05)。[结论]罗格列酮、二陈汤、苓桂术甘汤能改善大鼠肝脏脂肪变性,降低TNF-α和NF-κB蛋白表达。软脂酸可以明显抑制肝细胞增殖率,罗格列酮与苓桂术甘汤含药血清能拮抗软脂酸对肝细胞增殖的抑制。苓桂术甘汤通过抑制NF-κB蛋白表达,减少TNF-α表达,从而减轻肝脏炎性损伤,进而达到治疗NASH的作用。