sincalide and Myocardial-Infarction

This study aimed to investigate the effect and mechanism of miR-26a-5p on cardiomyocyte injury induced by hypoxia/reoxygenation (H/R).After construction of an H/R model in rat cardiomyocyte H9c2 cells, miR-26a-5p in the cells was interfered with (cells transfected with miR-26a-5p inhibitor) or overexpressed (cells transfected with a miR-26a-5p mimics). The viability and the apoptosis rate of cells in each group were detected using CCK-8 and flow cytometry; the relationship between miR-26a-5p and WNT5A was verified by a dual-luciferase reporter assay; the expression of miR-26a-5p, WNT5A, cleavedcaspase3 and Wnt/β-catenin signaling pathway-related proteins in each group was detected using qRT-PCR or Western blot; LDH release, SOD, and GSH-PX activities in each group were detected by kit.In the H/R group, the expression level of miR-26a-5p was significantly decreased, whereas the expression level of WNT5A was significantly increased. The activity of the Wnt/β-catenin signaling pathway was up-regulated; the level of LDH released was significantly increased; and activities of SOD and GSH-PX were significantly decreased. The aforementioned changes resulted in decreased cell activity and increased apoptosis rate. The overexpression of miR-26a-5p could reduce the expression level of WNT5A, the activity of the Wnt/β-catenin signaling pathway, and the apoptosis rate and restore the cell viability.These results suggest that miR-26a-5p can target WNT5A and thus, inhibit the Wnt/β-catenin signaling pathway activity, inhibiting H/R-induced cardiomyocyte injury and apoptosis.

Topics: Acute Disease; Animals; Apoptosis; Cell Survival; Flow Cytometry; Glutathione Peroxidase; Hypoxia; MicroRNAs; Models, Animal; Myocardial Infarction; Myocardial Reperfusion Injury; Myocytes, Cardiac; Protective Factors; Rats; Sincalide; Superoxide Dismutase; Transfection; Up-Regulation; Wnt Signaling Pathway; Wnt-5a Protein

Myocardial infarction (MI) increases myocardial fibrosis (MF) and subsequent cardiac remodeling. Cholecystokinin octapeptide (CCK-8) is expressed in cardiomyocytes and plays an important role in cardiovascular regulation. In this study, we intend to use a rat model of myocardial infarction to evaluate the effects of CCK-8 on myocardial fibrosis and cardiac remodeling.. Male Sprague-Dawley rats were separated into 3 groups: sham operation, MI + NaCl, and MI + CCK-8. All rats were subjected to left coronary artery ligation to induce MI or sham operation and then treated with CCK-8 or saline for 28 days. After 4 weeks, echocardiography was performed to assess cardiac function and myocardial fibrosis was evaluated using H&E and Masson's Trichrome-stained sections. The levels of BNP, CCK-8 in the plasma of all rats were detected by ELISA; RNA sequencing (RNA-seq) analysis was also adapted to detect differentially expressed genes in myocardial tissues of each group. Myocardial expression of fibrosis markers was analyzed by western blotting, immunohistochemistry and qRT-PCR.. CCK-8 was demonstrated to improve left ventricular function and results of H&E staining, Masson's trichrome staining, immunohistochemistry and western blotting showed that CCK-8 attenuated MF. Gene expression profiles of the left ventricles were analysed by RNA-seq and validated by qRT-PCR. Cardiac fibrosis genes were downregulated by CCK-8 in the left ventricle.. CCK-8 can alleviate fibrosis in the noninfarcted regions and delay the left ventricular remodeling and the progress of heart failure in a MI rat model.

Topics: Animals; Cardiomyopathies; Disease Models, Animal; Echocardiography; Gene Expression Regulation; Heart Ventricles; Male; Myocardial Infarction; Myocytes, Cardiac; Natriuretic Peptide, Brain; Rats; Rats, Sprague-Dawley; RNA-Seq; Sincalide; Ventricular Remodeling

Luhong formula (LHF)-a traditional Chinese medicine containing Cervus nippon Temminck, Carthamus tinctorius L., Cinnamomum cassia Presl, Codonopisis pilosula( Franch.) Nannf., Astragalus membranaceus ( Fisch.) Bge. var. mongholicus ( Bge.) Hsiao, Lepidium apetalum Willd-is used in the treatment of heart failure.. To investigate the antifibrotic efficacy of LHF in a myocardial infarction-induced rat model of heart failure and to determine its mechanism of action.. Myocardial infarction was induced in rats by coronary artery ligation, and cardiac fibroblasts were isolated. Neonatal rat cardiomyocytes (NRCMs) were isolated from 2 to 3-day-old Sprague-Dawley male rats, and cardiomyocyte hypertrophy was induced by isoprenaline. Histological examination was carried out to estimate the degree of myocardial fibrosis. Expression of gp130/JAK2/STAT3 pathway proteins was measured by western blot. The mRNA levels of downstream genes of gp130/JAK2/STAT3 pathway (i.e., CTGF, TSP-1, and TIMP1) were determined by RT-PCR; while CTGF, TSP-1, and TIMP1 protein levels were measured by ELISA. To investigate paracrine effects, cell proliferation and collagen synthesis was measured after treating cardiac fibroblasts with the conditioned media from isoprenaline-treated NRCMs.. Histopathological changes showed that LHF inhibited myocardial fibrosis in heart failure rats. Treatment with LHF up-regulated gp130, JAK2, and STAT3 protein expression in heart tissue, and down-regulated CTGF, TSP-1, and TIMP1 gene expression. Isoprenaline-treated NRCMs displayed lower expression of the gp130, JAK2, and STAT3 pathway proteins and higher secretion of its downstream signaling molecules (CTGF, TSP-1, TIMP1). LHF inhibited cardiac fibroblast proliferation and collagen synthesis after treatment with the conditioned media from isoprenaline-treated NRCMs.. LHF treatment attenuates myocardial fibrosis in vivo. LHF inhibits cardiac fibroblasts proliferation and collagen synthesis in a paracrine manner by activating the gp130/JAK2/STAT3 pathway in cardiomyocytes, thereby inhibiting the secretion of downstream profibrogenic cytokines.

Topics: Animals; Cytokine Receptor gp130; Drugs, Chinese Herbal; Fibrosis; Heart Failure; Janus Kinase 2; Lysosomal Membrane Proteins; Male; Medicine, Chinese Traditional; Myocardial Infarction; Myocytes, Cardiac; Paracrine Communication; Rats; Rats, Sprague-Dawley; Signal Transduction; Sincalide; STAT3 Transcription Factor