High-throughput Sequencing Analysis Of LncRNA And MRNA Of Huiyang Shengji Recipe In Regulating Diabetic Anal Leakage Postoperative Wounds

Objective: To study the specific expression of LncRNA and mRNA in wound of patients with diabetic anal fistula after operation with Huiyang Shengji formula,and analyze its mechanism of action.Methods:Six patients with type 2 diabetes mellitus were selected from the Second Affiliated Hospital of Hunan University of traditional Chinese medicine(Hunan Province Hospital of traditional Chinese Medicine).The control group was taken from the granulation tissue(A1,A2,A3,A4,A5,A6)with a diameter of about 0.5cm in the center of the wound after the operation.The experimental group was taken from the new granulation tissue(C1,C2,C3,C4,C5,C6)with a diameter of about 0.5cm in the same patient after 10 days of treatment,which was immediately frozen in a dry ice box and handed over to Shanghai Jingneng Biotechnology Co.,Ltd.for application of Illumina Hiseq sequencer completed the high-throughput sequencing,and screened the differentially expressed LncRNA and mRNA according to the sequencing results.Go analysis and KEGG analysis were carried out.Results:1792 differential LncRNAs were screened,including 646up-regulated genes and 1146 down regulated genes.Among 11778 differentially expressed genes,3730 were up-regulated and 8048 were down regulated.At the same time,we screened out three LncRNAs(ENST0000619449,ENST0000429829,ENST0000450589)related to this project.All the three indexes were verified by Q-PCR,and finally ENST0000450589 passed the verification.The down-regulation of LncRNA-cis,LncRNA-trans and mRNA was significant,and the function of go was mainly enriched in skin development,epidermal development,keratosis,keratinocyte differentiation and epidermal cell differentiation;the up-regulation of LncRNA-cis,LncRNA-trans and mRNA was significant,and the function of go was mainly enriched in cell,organelle,neuron and organelle.KEGG pathway analysis screened three pathways which are most relevant to this project,namely endocytosis pathway,PPAR signaling pathway and focal adhesion pathway.Conclusion: LncRNA and mRNA play an important role in wound healing.One lncrna(ENST00004050589)related to this project and verified by Q-PCR was screened.The regulation of Huiyang Shengji Recipe on LncRNA and mRNA in wound healing of diabetic anal fistula mainly focused on epidermal development,skin development,keratinization,keratinocyte differentiation,epidermal cell differentiation,etc.The LncRNA and mRNA of Huiyang Shengji recipe applied in diabetic anal fistula wound may participate in regulating wound healing through endocytosis pathway,PPAR signaling pathway and focal adhesion pathway.