The LncRNA Expression Analysis Of Jiangtangtiaozhi Prescription In Patients With Obese Type 2 Diabetes Mellitus And Dyslipidemia

In recent years,obese type 2 diabetes mellitus and dyslipidemia became a very common clinical combined disease with the rapid growth in the incidence of diabetes.The treatment is to reduce blood sugar,to decrease blood lipids and to lose Weight.Traditional Chinese medicine treatment is always multi-channel and multi-target.It can intervene this complex disease by reducing blood glucose,blood lipids,body weight and other abnormal clinical indicators simultaneously.It is a combined effect on this disease,and there is a distinct advantage in the regulation of the patients’ overall symptoms.Research on Long noncoding RNA（LncRNA）is a central issue recently,and researchs of the relationship between diabetes and LncRNA are being gradually unfolded.However,there is no report yet of LncRNA expression analysis on Traditional Chinese medicine interventions in type 2 diabetes.Objective:Previous studies have confirmed the effectiveness of Jiangtangtiaozhi Prescription on reducing blood sugar,blood lipids and body weight.This study will analyze the LncRNA differentially expression in patients with obese type 2 diabetes mellitus and dyslipidemia compared to healthy people.Furthermore,we will analyze the differences of LncRNA expression on comparison of before and after the intervention of Jiangtangtiaozhi Prescription,and also explore the possible mechanism of this prescription on treating obese type 2 diabetes mellitus and dyslipidemia.Methods:We chose 12 patients with obese type 2 diabetes mellitus and dyslipidemia（syndrome of excess of gastrointestinal heat）who were treated by Jiangtangtiaozhi Prescription of 24 weeks,while 12 cases included in the healthy control group.First,we select 6 cases in each group（disease group before treatment,disease group after treatment and healthy control group）to start the research of LncRNA microarray.According to the differentially expressions of LncRNAs and mRNAs,we secondly do GO analysis,Pathway analysis,and find out some target LncRNAs as well as their associated mRNAs.Then,each group will be expanded to 12 cases in the qPCR validation on these target LncRNAs and mRNAs.Finally,a detailed analysis of the differences on disease group before treatment vs.healthy control group and disease group after treatment vs.before will be managed.In addition,we will focus on disease-related pathways,and will analyze the correlation of differentially expressions of target IncRNAs with the clinical indicators.Results:1.Disease group before treatment vs.healthy control group（1）There are 557 upregulated LncRNAs,273 downregulated LncRNAs,491 upregulated mRNAs and 1639 downregulated mRNAs.（2）GO analysis:upregulated GO items:molecular function（MF）40 items,biological processes（BP）144 items,cell components（CC）44 items.The downregulated GO items:MF 106 items,BP 541 items,CC 126 items.（3）Pathway analysis:There were 6 pathways enriched by the upregulated mRNAs,sort by enrichmentscore:Serotonergic synapse synaptic,Protein digestion and absorption,Synaptic vesicle cycle,Fatty acid degradation,Calcium signaling pathway,Basal cell carcinoma.There were 39 pathways enriched by the downregulated mRNAs,top 10 sort by enrichmentscore:Spliceosome,Pyruvate metabolism,Cysteine and methionine metabolism,RNA degradation,Glycolysis/Gluconeogenesis,Carbon metabolism,Ubiquitin mediated proteolysi,Protein export,Protein processing in the endoplasmic reticulum,RNA transport.（4）qPCR validation:the expression of XLOC-005590 and HNF1A-AS1 as target LncRNA were raised,and the gene chips had the same sexual results.2.Disease group after treatment vs.before（1）There are 128 upregulated LncRNAs,32 downregulated LncRNAs,45 upregulated mRNAs and 140 downregulated mRNAs.（2）GO analysis:upregulated GO items:molecular function（MF）24 items,biological processes（BP）79 items,cell components（CC）14 items.The downregulated GO items:MF 14 items,BP 57 items,CC 51 items.（3）Pathway analysis:There was a pathway enriched by the upregulated mRNAs:Peroxisome.There were 5 pathways enriched by the downregulated mRNAs,sort by enrichmentscore:Basal transcription factors,Other types of O-glycan biosynthesis other types O-glycan biosynthesis,Insulin signaling pathway of insulin signaling pathway,Cysteine and methionine metabolism cysteine and methionine metabolic,Glycine,serine and threonine metabolism of glycine and serine and threonine metabolism.（4）qPCR validation:Referring to the healthy control group,XLOC₀05590 was upregulated and the downstream gene（ECI2）was downregulated in disease group before treatment.However,after 24 weeks’ intervention by Jiangtangtiaozhi Prescription,XLOC₀05590 was downregulated and ECI2 was upregulated compared with the group of treatment before（0 weeks）.The differences were statistically significant multiples（P<0.05）.Conclusions:1.Compared Disease group before treatment with healthy control group:The significant differences between obese type 2 diabetes mellitus and dyslipidemia patients with healthy people were showed in fatty acid degradation pathway significantly upregulated and pathways of glycolysis/gluconeogenesis and pyruvate metabolism significantly downregulated.We speculated XLOC-005590,HNF1A-AS1 will be the potential biomarkers of obese type 2 diabetes mellitus and dyslipidemia.2.Compared Disease group after treatment with Disease group before treatment:we speculated XLOC-005590 as a potential therapeutic target biomarkers while patients with obese type 2 diabetes mellitus and dyslipidemia were treated by Jiangtangtiaozhi Prescription.One of the possible mechanism for this Prescription is regulating the expression of XLOC-005590 which will reverse regulate ECI2,furthermore,ECI2 may play a role by affecting peroxisomes and fatty acid degradation pathway.