Spironolactone Protects against Diabetic Cardiomyopathy in Streptozotocin-Induced Diabetic Rats

Abstract

Spironolactone (SPR) has been shown to protect diabetic cardiomyopathy (DCM), but the specific mechanisms are not fully understood. Here, we determined the cardioprotective role of SPR in diabetic mice and further explored the potential mechanisms in both in vivo and in vitro models. Streptozotocin- (STZ-) induced diabetic rats were used as the in vivo model. After the onset of diabetes, rats were treated with either SPR (STZ + SPR) or saline (STZ + NS) for 12 weeks; nondiabetic rats were used as controls (NDCs). In vitro, H9C2 cells were exposed to aldosterone, with or without SPR. Cardiac structure was investigated with transmission electron microscopy and pathological examination; immunohistochemistry was performed to detect nitrotyrosine, collagen-1, TGF-β1, TNF-α, and F4/80 expression; and gene expression of markers for oxidative stress, inflammation, fibrosis, and energy metabolism was detected. Our results suggested that SPR attenuated mitochondrial morphological abnormalities and sarcoplasmic reticulum enlargement in diabetic rats. Compared to the STZ + NS group, cardiac oxidative stress, fibrosis, inflammation, and mitochondrial dysfunction were improved by SPR treatment. Our study showed that SPR had cardioprotective effects in diabetic rats by ameliorating mitochondrial dysfunction and reducing fibrosis, oxidative stress, and inflammation. This study, for the first time, indicates that SPR might be a potential treatment for DCM.

Phospholipase C inhibits apoptosis of porcine oocytes cultured in vitro

“…The RNA concentration was determined using a spectrophotometer (NanoDrop 2000c; Thermo Fisher Scientific, Waltham, MA) with an absorbance of 260 nm. Total RNA was converted to complementary DNA using 5 × All‐In‐One RT MasterMix (ABM, Vancouver, Canada) . Primers (Table ) were synthesized by Sangon Biotech (Shanghai, China).…”

Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro

“…The granulosa cells were treated for 2 h, 4 h, 8 h, 12 h, 24 h and 48 h which was used for detecting PLCB1 mRNA expression or for 4 h which was used for detecting other genes, and then total RNA was extracted from the cells using TRIzol reagent (Takara, Kyoto, Japan) as described [21]. RNA concentration was determined using a spectrophotometer (NanoDrop 2000c, Thermo Scientific, Waltham, Massachusetts, USA) by absorbance at 260 nm [22]. Then, 1 μg of total RNA was converted to cDNA using 5 × All-In-One RT MasterMix (Abm, Vancouver, Canada) according to the manufacturer’s protocol [22].…”

“…RNA concentration was determined using a spectrophotometer (NanoDrop 2000c, Thermo Scientific, Waltham, Massachusetts, USA) by absorbance at 260 nm [22]. Then, 1 μg of total RNA was converted to cDNA using 5 × All-In-One RT MasterMix (Abm, Vancouver, Canada) according to the manufacturer’s protocol [22]. Primers were synthesized by Sangon Biotech (Shanghai, China), and the primer sequences used for genes studied are listed in Table 1.…”

Novel Insights Into Molecular Mechanism of Mitochondria in Diabetic Cardiomyopathy

“… Fang et al (2018) confirmed that resveratrol might alleviate cardiac oxidative stress, mitochondrial impairment, and myocardial fibrosis in diabetes induced by high glucose. Liu et al (2018) explored the protection of spironolactone against DCM in STZ-induced diabetic rats and concluded that its cardioprotective effects were due to improving mitochondrial dysfunction and reducing cardiac fibrosis, oxidative stress, and inflammation ( Liu et al, 2018 ). Hu et al (2019) demonstrated that the reconstruction of Mfn2 restored mitochondrial membrane potential, inhibited mitochondrial oxidative stress, and ameliorated mitochondrial function in cardiomyocytes treated by high glucose and high-fat through facilitating mitochondrial fusion.…”