摘要
目的基于高通量测序及维恩图预测美洲大蠊提取液促进糖尿病肛漏创面愈合的顺、反式lnc RNA及Pathway。方法选取湖南中医药大学第二附属医院收治的糖尿病肛漏患者术后创面12例,利用高通量测序技术检测6例术后创面组(A组)和6例术后创面应用美洲大蠊提取液创面组(B组)中lnc RNAs和m RNAs表达,进行GO分析和KEGG通路分析,构建lnc RNA-m RNA的共表达网络图,并通过Cis-及Trans-预测与创面愈合相关lnc RNA。结果实验组差异表达的lnc RNAs2242个(上调649个,下调1593个),m RNAs有13186个(上调5162个,下调8024个)。GO分析发现差异表达的lnc RNA-cis主要富集在表皮细胞分化、上皮细胞分化、细胞代谢过程的调节等;差异表达的lnc RNA-trans主要富集在细胞代谢过程、蛋白质结合、细胞内部分等。通过KEGG通路分析,筛选出与本研究相关的3条通路:IBD、MAPK signalingpathway、AMPK signaling pathway;lnc RNA靶标基因预测中最终筛选出3个lnc RNA(ENST00000443364、ENST00000576797、ENST00000620167),进行PCR验证,lnc RNAENST00000443364、ENST00000576797与芯片结果一致。结论美洲大蠊提取液可能通过lnc RNAENST00000443364、ENST00000576797顺式及反式调控靶标基因,影响表皮细胞分化、上皮细胞分化、细胞代谢过程的调节等功能以及影响IBD、MAPK signalingpathway、AMPK signaling pathway促进糖尿病肛漏创面愈合。
Objective To predict the cis and trans lnc RNAs and Pathway of Periplaneta americana extracts promotingdiabetic anal leakage wound healing based on high-throughput sequencing and Venn diagram.Methods Twelvepostoperative trauma cases of diabetic anal leakage patients admitted to the Second Affiliated Hospital of HunanUniversity of Chinese Medicine were selected,and the expression of lnc RNAs and m RNAs in 6 postoperative traumagroups(group A)and 6 postoperative trauma groups with the application of American cockroach extract(group B)weredetected by high-throughput sequencing technology,and GO analysis and KEGG pathway analysis were performed toconstruct a lnc RNA-m RNA co-expression network maps,and predicted lnc RNAs associated with trauma healing byCis-and Trans-.Results There were 2242 differentially expressed lnc RNAs(649 up-regulated and 1593 down-regulated)and 13186 m RNAs(5162 up-regulated and 8024 down-regulated)in the experimental group.GO analysisrevealed that the differentially expressed lnc RNA-cis was mainly enriched in epidermal cell differentiation,epithelialcell differentiation,regulation of cellular metabolic processes,etc.The differentially expressed lnc RNA-trans weremainly enriched in cellular metabolic processes,protein binding,intracellular fraction,etc.Through KEGG pathwayanalysis,3 pathways related to this study were screened:IBD、MAPK signaling pathway、AMPK signaling pathway;3 lnc RNAs(ENST00000443364,ENST00000576797,ENST00000620167)were finally screened in lnc RNA target geneprediction.PCR was performed to verify that lnc RNAs ENST00000443364 and ENST00000576797 were consistent withthe microarray results.Conclusion The extract of Periplaneta americana may regulate target genes in cis and trans vialnc RNAENST00000443364 and ENST00000576797,affect epidermal cell differentiation,epithelial cell differentiation,regulation of cell metabolism and other functions,as well as IBD、MAPK signaling pathway、AMPK signaling pathway topromote diabetic anal leakage wound healing.
作者
李心甜
马乐
耿越飞
沈咏梅
曾娟妮
Li Xintian;Ma Le;Geng Yuefei;Shen Yongmei;Zeng Juanni(The Second Affiliated Hospital of Hunan University of Chinese medicine,Changsha 410005,China;The First Affiliated Hospital of Hunan University of Chinese medicine,Changsha 410007,China;Sichuan Key Laboratory of Medical American Cockroach,Chengdu 610000,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2021年第4期1147-1155,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(81774325):基于CNC生物网络图及在体、离体实验差析养阴、回阳机制十三五全国中医药创新人才支助,负责人:曾娟妮。
