Influence Of Jinhu Gushen Chinese Medicine Formula On Peripheral Blood MiRNA Expression Profile

Objective:We used Small RNA library sequencing combined with clinical features and laboratory results,to analyze the differences in peripheral blood mi RNA expression profiles of patients with diabetic kidney disease between patients with type 2 diabetes without diabetic kidney disease,and effect of Jinhu Gushen Chinese medicine formula on peripheral blood mi RNA expression profile in peripheral blood of patients with diabetic kidney disease.The mechanism and target of drug were studied by bioinformatics analysis,which provides a theoretical basis for the treatment of diabetic kidney disease with traditional Chinese medicine.Methods:10 patients with diabetic kidney disease and which e GFR≥60ml/min/1.73m²,UACR>300 mg/g admitted to the Endocrinology Department of Affiliated Hospital of Qingdao University were selected as the traditional Chinese medicine treatment group.The clinical data such as UACR,Hb A1c and complications,and peripheral blood RNA specimens were collected at the baseline and after 6 months of medication.And 10 patients with type 2 diabetes without diabetic kidney disease（e GFR≥90ml/min/1.73m²,UACR<30 mg/g）were selected as the control group,and the clinical data and peripheral blood RNA samples were collected only at the baseline.The RNA samples were sent to Novegene Bioinformatics Technology Company for database construction and screened out the differentially expressed mi RNAs between the traditional Chinese medicine treatment group with the control group and the traditional Chinese medicine treatment group after treatment with before.The target genes’pathways of these mi RNAs were analysed by GO and KEGG.Combined with clinical data and bioinformatics analysis results,the mechanism and targets of Jinhu Gushen Chinese medicine formula were explored.Results:The median UACR of the control group and the traditional Chinese medicine treatment group was 7.98（5.16,9.93）mg/g,871.71（453.23,2480.00）mg/g,respectively.The UACR of the traditional Chinese medicine treatment group was significantly higher than that of the control group（p<0.001）.After 6 months of Jinhu Gushen Chinese medicine formula intervention,the UACR in the traditional Chinese medicine treatment group decreased significantly（p=0.005）,with a median of 403.42（182.84,1489.01）mg/g,and the median decrease rate of 53.72%.A total of 80 mi RNAs（40 were up-regulated and 40 were down-regulated）（p<0.05 and|log₂ foldchange|>1）were differentially expressed in the traditional Chinese medicine treatment group compared with the control group.And there were 36 differentially expressed mi RNAs（18 up-regulated and 18 down-regulated）（p<0.05 and|log₂ foldchange|>1）in the traditional Chinese medicine treatment group after treatment compared with those before treatment.Among them,there were 8 mi RNAs whose expression was up-regulated in the traditional Chinese medicine treatment group compared with the control group and down-regulated after treatment,namely mi R-1262,mi R-4772-3p,mi R-1298-5p,mi R-1260b,novel-599,mi R-493-5p,mi R-532-5p,mi R-28-3p.And there were 6 mi RNAs whose expression was down-regulated in the traditional Chinese medicine treatment group compared with the control group and up-regulated after treatment,namely mi R-219a-2-3p,mi R-219b-5p,mi R-433-3p,mi R-133a-3p,mi R-499a-5p,mi R-499b-3p.KEGG pathway enrichment analysis showed that MAPK,PI3K-Akt,Ras,Rap1,Ca²⁺,HIF-1 and other signaling pathways,as well as endocytosis,autophagy,mitophagy,metabolic pathways,etc.were closely related to the progression of DKD.The MAPK,PI3K-Akt,Ras,Rap1,Ca²⁺,TRP and other signaling pathways,as well as endocytosis,autophagy,mitophagy,metabolic pathways,insulin resistance,etc.were the mechanisms of Jinhu Gushen Chinese medicine formula.Conclusion:1.Jinhu Gushen Chinese medicine formula can significantly reduce urinary protein excretion in patients with diabetic kidney disease.2.Jinhu Gushen Chinese medicine formula reverses the expression of mi R-28-3p,mi R-493-3p,mi R-133a-3p,mi R-499a-5p,mi R-499b-3p and other mi RNAs in patients with diabetic kidney disease.3.Jinhu Gushen Chinese medicine formula may reduce UACR and delay the progression of DKD by affecting MAPK,PI3K/Akt,Rap1/Ras,TRP and other signaling pathways,as well as metabolic pathways,mitophagy,autophagy,and endocytosis.