RESUMO
Ischemic heart disease (IHD) is one of the world's leading causes of human death. Kaempferol (Kae) was proved to have anti-inflammatory, antioxidant, and anticancer effects. Such properties suggested that it might play protective roles in IHD. In this study, we have attempted to disclose the potential regulating mechanisms of Kae in primary cardiomyocytes and H9c2 cells.Cells were first stimulated by oxygen-glucose deprivation (OGD) and then exposed to Kae. CCK-8 assay and flow cytometry were used to examine cell characteristics. Quantitative reverse-transcription polymerase chain reaction was utilized to test the expression levels of miR-15b and TLR4. Afterward, cell transfection, dual-luciferase activity assay, and western blot were used to explore the potential mechanisms.OGD treatment suppressed cell viability, whereas it enhanced cell apoptosis. Besides, OGD treatment enhanced the expression of apoptosis-associated proteins. Kae exposure, however, attenuated the effects that OGD-induced. Further experiments showed that Kae exposure promoted down-regulation of miR-15b, Bcl-2 and TLR4 were a target of miR-15b. Moreover, Kae enhanced the expression of key factors involved in PI3K/AKT and Wnt/ß-catenin pathways, whereas miR-15b mimic reversed the Kae-triggered effects.This investigation revealed that Kae diminished OGD-triggered cell damage through down-regulating miR-15b expression via activating PI3K/AKT and Wnt3a/ß-catenin pathways.
Assuntos
Quempferóis/uso terapêutico , MicroRNAs/metabolismo , Isquemia Miocárdica/tratamento farmacológico , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos , Quempferóis/farmacologia , Isquemia Miocárdica/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Ratos Wistar , Via de Sinalização Wnt , beta Catenina/metabolismoRESUMO
Background/Aims: Myocardial ischemia (MI) is a serious threat to human health. Circular RNAs (circRNAs) play an important role in many diseases including MI. The effect and mechanism of circDENND2A in MI have not been studied.Methods: We used oxygen glucose deprivation (OGD) treatment to simulate MI in vitro. We detected circDENND2A and microRNA (miR)-34a levels by RT-qPCR. The transfection process used INTERFER and jetPRIME. Cell growth indexes including viability, apoptosis, and migration were detected by CCK8, flow cytometry, and transwell assays, respectively. In addition, the Bax, Cleaved-Caspase-3, matrix metalloproteinase (MMP)-2, MMP-9 and pathway-related protein levels were tested by Western blot.Results: OGD upregulated circDENND2A expression in H9c2 cells. Overexpression of circDENND2A enhanced cell viability and migration but declined apoptosis under OGD. Silenced circDENND 2A played the opposite effects. circDENND2A negatively regulated miR-34a. miR-34a overexpression weakened the protective effects of circDENND2A in OGD-injury. Moreover, we considered circDENND2A and miR-34a may work via ß-catenin and Ras/Raf/MEK/ERK pathways.Conclusion: circDENND2A overexpression enhanced OGD-inhibited cell viability and migration but declined OGD-promoted apoptosis by downregulating miR-34a and via ß-catenin and Ras/Raf/MEK/ERK pathways.
Assuntos
Apoptose/genética , Citoproteção , Glucose/deficiência , MicroRNAs/metabolismo , Oxigênio/metabolismo , RNA Circular/metabolismo , Idoso , Animais , Linhagem Celular , Movimento Celular/genética , Proliferação de Células/genética , Citoproteção/genética , Regulação para Baixo/genética , Feminino , Inativação Gênica , Humanos , Sistema de Sinalização das MAP Quinases , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Infarto do Miocárdio/genética , RNA Circular/genética , Ratos , Regulação para Cima/genética , beta Catenina/metabolismoRESUMO
BACKGROUND: Hypertension has become prevalent among young and middle-age individuals. Many studies have identified a variety of risk factors for cardiovascular diseases, yet there have been few reports focusing on the young and middle-age hypertensive population. OBJECTIVE: To investigate the epidemiological characteristics of conventional risk factors of premature coronary artery disease (PCAD) in patients with hypertension. METHODS: The clinical and laboratory data of 267 hypertensive patients with PCAD and 96 hypertensive patients without any visible coronary disease according to angiography were compared. Potential coronary risk factors were analyzed using logistic regression. RESULTS: The PCAD group had lower levels of high-density lipoprotein cholesterol (HDL-C) (P<0.05). Multivariate logistic analysis showed positive family history, low HDL-C, hypertriglyceridemia, duration of diabetes mellitus and male sex were significantly associated with PCAD (P<0.05), with ORs of 12.317, 3.267, 2.894, 1.140 and 0.088, respectively. Plasma renin activities in PCAD patients were significantly higher than in control hypertensive patients (P=0.027), but there was no significant difference in angiotensin II and aldosterone levels between the two groups. CONCLUSION: Low HDL-C and hypertriglyceridemia are important coronary risk factors in Chinese individuals with hypertension.