Study On Regulating Mechanism Of Yanghua Decoction On Serum Exosomal MiRNA In Patients With Yang Deficiency Syndrome In Acute Ischemic Stroke

Objective: In this paper,we use exosomal sequencing technology to explore the regulatory mechanism of Yanghua Decoction on serum exosomal miRNA in patients with acute ischemic stroke(Acute Ischemic Stroke,AIS).Methods: The subjects including the 16 AIS patients and 8 healthy subjects.To be extend,the AIS patients were randomly divided into the treatment and control group with 8 cases respectively.Besides,the treatment group was treated with Yanghua Decoction on the basis of the control group during the observation period of 7 days.Firstly,the NIHSS scale,ADL scale and TCM syndrome score were performed on AIS patients before and after treatment.All kinds of the healthy subjects collected serum samples on day 0 of enrollment,and AIS patients collected serum samples on day 7 to isolate and identify exosomes.Moreover,the small RNA sequencing technology was used to sequence the miRNAs in exosomes to screen out the differentially expressed exosomes miRNAs after treatment with Yanghua Decoction.Finally,we use the TargetScan,miRDB and miRWalk to predict the target genes of the differential miRNAs screened out to analyze the function of the target genes and related pathways.In brief,the regulation mechanism of Yanghua Decoction on serum exosomal miRNA in patients with AIS,which provide reference for clinical research of AIS.Results:1.The analysis results of the serum exosomal miRNA expression:(1)Compared with the control group,there are 25 miRNAs differentially expressed in serum exosomes of patients in the treatment group,of which 12 are up-regulated and 13 are down-regulated;(2)In the treatment group,there are 25 miRNAs differentially expressed in serum exosomes of patients compare with the control group,of which 12 are up-regulated and 13 are down-regulated;(3)We could determine that the three most significant exosomal miRNAs in the serum of AIS patients with Yang-deficiency syndrome regulated by Yanghua Decoction are miR-9-3p,miR-9-5p,and miR-124-3p.2.The bioinformatics analysis results of the serum exosomes miR-9-3p/5p and miR-124-3p:(1)The target gene prediction: A total of 475 target genes of 3 exosomal miRNAs are predicted.(2)The functional enrichment analysis of target genes: The functional analysis of the target genes of three exosomal miRNAs revealed that multiple biological processes with the variety of biological functions including the intercellular junctions,two-cell tight junction components,transcription factor activity,RNA polymerase II transcription factor binding,etc.,exert a variety of biological functions.(3)Target gene pathway analysis: 3 exosomal miRNA target genes enriched a total of 12 pathways.In particular,the signal transduction pathways include cGMP-PKG signaling pathway,Apelin signaling pathway,insulin signaling pathway,etc.Apart from that the biochemical metabolic pathways include endocrine resistance and cellular senescence,Endocytosis,etc.3.The analysis results of the curative effect of Yanghua Decoction in the treatment of AIS patients with Yang-deficiency syndrome:The experimental results showed that the treatment and control group improved the scores of NIHSS scale,ADL scale and TCM syndrome scores after treatment of AIS patients.Moreover,the treatment group with better effect than the control group(P<0.05).Conclusion: The Yanghua Decoction combined with western medicine conventional treatment,which could improve the neurological deficit and the ability of daily living with AIS patients.Furthermore,Yanghua Decoction may down-regulate the expression of serum exosomes miR-9-3p/5p and miR-124-3p in patients with Yang-deficiency syndrome of AIS,which could better affect signal pathways like cGMP-PKG.