Study on the role of indole sulfate in inducing myocardial hypertrophy and fibrosis through organic anion transporter-3 and oxidative stress
10.3760/cma.j.cn431274-20221116-01189
- VernacularTitle:硫酸吲哚酚通过有机阴离子转运蛋白-3和氧化应激诱导心肌肥大及纤维化的作用
- Author:
Aizezi MAIMAITIAILI
1
;
Yukui DU
;
Yisireyili MAIMAITI
;
Jinjie SHAO
;
Jun LIU
;
Zonggang ZHANG
Author Information
1. 新疆维吾尔自治区人民医院心脏及泛血管医学诊疗中心心外一科，乌鲁木齐 830001
- Keywords:
Indoxyl sulfate;
Organic anion transport protein 3;
Cardiomyocyte;
Cardiac hypertrophy;
Fibrosis
- From:
Journal of Chinese Physician
2023;25(10):1484-1489
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the role of indoxyl sulfate (IS) in myocardial hypertrophy and fibrosis through organic anion transporter-3 (OAT-3) and oxidative stress in H9C2 cells.Methods:Rat myocardial cells (H9C2) were cultured and divided into four groups: Control group, IS group, siRNA negative control group (siOAT-3), and siOAT-3+ IS group. The control group was cultured routinely without IS stimulation. The IS group was stimulated with 250 μmol IS. The siRNA negative control group was transfected with 20 nmol/L OAT-3 siRNA, and the siOAT-3+ IS group was transfected with OAT-3 siRNA 48 hours later, then stimulated with IS for 24 hours. Real-time polymerase chain reaction (RT-PCR) was used to detect and analyze the mRNA expression levels of OAT-3, NADPH oxidase-4 (Nox-4), antioxidant proteins [nuclear factor E2 related factor 2 (Nrf-2), heme oxygenase 1 (HO-1)], myocardial hypertrophy markers [atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), β-myosin heavy chain gene (β-MHC)], and fibrosis markers [transforming growth factor β1 (TGF-β1), Smad homologous protein 3 (Smad-3), collagen type Ⅰ (Collagen Ⅰ)] in each group. H9C2 cells were further divided into three groups: Control group, IS group, and N-acetylcysteine (NAC) group. The NAC group was pre-treated with the antioxidant NAC before stimulation with 250 μmol IS for 24 hours. RT-PCR was used to detect the mRNA expression levels of the above indicators.Results:The mRNA expression level of OAT-3 in the IS group was significantly higher than that in the control group, while the mRNA expression levels of OAT-3 in the siOAT-3 group and the siOAT-3+ IS group were significantly lower than those in the IS group (all P<0.001). Compared with the Control group, the mRNA relative expression levels of Nox-4, ANP, BNP, β-MHC, TGF-β1, Smad-3, and Collagen Ⅰ in H9C2 cells of the IS group were significantly increased (all P<0.001), while the mRNA expression levels of Nrf-2 and HO-1 were significantly decreased (all P<0.001). Compared with the IS group, the mRNA relative expression levels of Nox-4, ANP, BNP, β-MHC, TGF-β1, Smad-3, and Collagen Ⅰ in H9C2 cells of the siOAT-3 group and the siOAT-3+ IS group were significantly lower (all P<0.001), while the mRNA expression levels of Nrf-2 and HO-1 were significantly increased (all P<0.001). Pre-treatment with NAC significantly inhibited the high expression of Nox-4, ANP, BNP, β-MHC, TGF-β1, Smad-3, and Collagen Ⅰ mRNA induced by IS (all P<0.001), while significantly increasing the mRNA expression levels of Nrf-2 and HO-1 (all P<0.01). Conclusions:IS promotes myocardial hypertrophy and fibrotic factor overexpression in H9C2 cells through OAT-3 and oxidative stress.
