AUTHOR=Wang Jie , Liu Lanchun , Liu Chao , Cheng Nuo , Mao Qiyuan , Chen Cong , Hu Jun , He Haoqiang , Hui Xiaoshan , Qu Peirong , Lian Wenjing , Duan Lian , Dong Yan , Liu Yongmei , Li Jun 

TITLE=Identification and analysis of differential miRNA–mRNA interactions in coronary heart disease: an experimental screening approach

JOURNAL=Frontiers in Cardiovascular Medicine

VOLUME=Volume 10 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/cardiovascular-medicine/articles/10.3389/fcvm.2023.1186297

DOI=10.3389/fcvm.2023.1186297

ISSN=2297-055X

ABSTRACT=Objective: This study aimed to screen the differential molecules of kidney deficiency and blood stasis syndrome in coronary heart disease by high-throughput sequencing, verify the expression level changes of miR-4685-3p and its regulated downstream C1QC, C4 and C5 by qPCR and ELISA, and to clarify that complement and coagulation cascade pathway may be the specific pathogenic pathway. Methods: Unstable angina pectoris patients with kidney deficiency and blood stasis (KDBS) syndrome of , patients with non-kidney deficiency blood stasis (NKDBS) syndrome and Nomal group were recruited respectively, and their clinical symptoms were analyzed. Illumina's NextSeq 2000 sequencing platform and FastQC software were used for RNA sequencing and quality control. DESeq software was used for differential expression analysis. qPCR and ELISA verification were performed on Differential Expression Analysis (DEG). Result: The differential gene expression profiles of 77 miRNA and 331 mRNA were selected. The GO enrichment analysis included 43 biological processes, 49 cell components, and 42 molecular functions. The KEGG enrichment results included 40 KEGG pathways. PCR results showed that, compared with the Nomal group, miR-4685-3p decreased in the CHD_KDBS group (P=0.001), and was lower than that in the CHD_NKDBS group. The downstream mRNA C1 regulated by miR-4685-3p showed an increasing trend in the CHD_KDBS group, which was higher than Nomal group (P=0.0019). The mRNA C4 and C5 in the CHD_KDBS group showed an upward trend, but the difference was not statistically significant. ELISA testing of proteins related to complement and coagulation cascade pathway. It was found that the expression level of C1 was significantly upregulated in the CHD_KDBS group compared with the Nomal group (P<0.0001), which higher than that in the CHD_NKDBS group(P<0.0001). The expression level of C4 and C5 in the CHD_KDBS group were significantly lower than Nomal group, and was lower than that in the CHD_NKDBS group (P<0.0001). Conclusion: The occurrence of CHD_KDBS might be related to the activation of complement and coagulation cascade pathway, in which miR-4685-3p decreases, its downstream C1QC had an up-regulated trend, and the expression levels of complement C4 and C5 decreased, which provided a research basis for the prevention and treatment of the disease.