目的 了解芥子酸（SA）对2型糖尿病小鼠心肌纤维化发生的影响及相关机制。方法 40只雄性C57BL/6J小鼠随机分为正常组（NOR）、2型糖尿病模型组（DM）、二甲双胍组（DM+MET, 200 mg/kg）、低剂量SA组（DM+SAL, 20 mg/kg）和高剂量SA组（DM+SAH, 60 mg/kg）。2型糖尿病模型以高脂日粮（HFD）联合链脲佐菌素（STZ,35 mg/kg）诱导,连续干预4 w。小鼠被处死后,计算心脏指数;HE和Masson染色分别观察心肌病理改变及纤维化程度;检测小鼠血清超氧化物歧化酶（SOD）、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶（GSH-Px）、丙二醛（MDA）、总胆固醇（TG）、甘油三脂（TC）及IL-1β、IL-6、IL-18和TNF-α水平;采用蛋白免疫印迹法检测心脏组织TGF-β1、α-SMA、Smad4、Smad7的蛋白表达水平;采用免疫组化法检测心肌α-SMA的原位表达状况。结果 与NOR组相比,DM组心肌组织形态结构紊乱,心脏指数升高（P<0.05）,心肌纤维化水平增高（P<0.05）,血清MDA、TG、TC、IL-1β、IL-6、IL-18及TNF-α水平明显上升（P<0.05）,心肌TGF-β1、α-SMA、Smad4蛋白表达增加（P<0.05）,而血清SOD、CAT、GSH-Px及心肌Smad7蛋白表达水平明显下降（P<0.05）;与DM组相比,DM+MET组、DM+SAL组和DM+SAH组心脏指数明显下降（P<0.05）,心肌纤维化水平下降（P<0.05）,血清MDA、TG、TC、IL-1β、IL-6、IL-18、TNF-α水平显著降低（P<0.05）,TGF-β1、α-SMA、Smad4蛋白表达明显下降（P<0.05）,而血清SOD、CAT、GSH-Px及心肌Smad7水平明显提高（P<0.05）。结论 SA干预可抑制2型糖尿病小鼠心肌纤维化的发生,可能与调控TGF-β/Smads信号通路有关。
Abstract
Objective To investigate the effect of sinapic acid (SA) on myocardial fibrosis in type 2 diabetic mice and its related mechanism. Methods Forty C57BL/6J mice were divided into the normal group (NOR), the type 2 diabetes model group (DM), the metformin group (DM+MET, 200 mg/kg), the low-dose sinapic acid group (DM+SAL, 20 mg/kg), and the high-dose sinapic acid group (DM+SAH, 60 mg/kg). The type 2 diabetes model was induced with a high-fat diet (HFD) in combination with the injection of streptozotocin (STZ, 35 mg/kg). The intervention lasted four weeks. All animals were sacrificed and the cardiac index was calculated. Myocardial pathological changes and histological fibrosis levels were evaluated through HE and Masson's staining. The commercial kits were used to assess oxidative stress indicators, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA), in addition to serum cholesterol (TC) and triglycerides (TG), and inflammation indicators such as interleukin-1β (IL-1β), IL-6, IL-18, and tumor necrosis factor-α (TNF-α). Immunohistochemistry was used to measure the levels of cardiac α-SMA, while Western-blot was employed to detect the levels of TGF-β1, α-SMA, Smad4, and Smad7 in the cardiac tissues. Results In comparison to the NOR group, the DM group exhibited disorganized myocardial morphology, increased cardiac index (P<0.05), augmented myocardial fibrosis level (P<0.05), elevated levels of serum MDA, TG, TC, IL-β1, IL-6, IL-18, and TNF-α (P<0.05), and up-regulated expressions of myocardial TGF-β1, α-SMA, and Smad4 (P<0.05). Conversely, there were significantly decreased levels of serum SOD, CAT, GSH-Px, and myocardial expression of Smad7 (P<0.05). Compared to the DM group, the cardiac index of mice in the DM+MET, DM+SAL and DM+SAH groups was significantly lower (P<0.05), myocardial fibrosis was reduced (P<0.05), serum levels of MDA, TG, TC, IL-1β, IL-6, IL-18, and TNF-α were significantly decreased (P<0.05), and TGF-β1, α-SMA, and Smad4 protein expressions were significantly reduced (P<0.05). Meanwhile, serum levels of SOD, CAT, GSH-Px, and myocardial Smad7 expression were significantly increased (P<0.05). Conclusion Sinapic acid alleviates myocardial fibrosis in T2DM mice, which is related to the regulation of the TGF-β/Smads signaling pathway.
关键词
芥子酸 /
2型糖尿病 /
心肌纤维化 /
小鼠
Key words
sinapic acid /
type 2 diabetes /
myocardial fibrosis /
mice
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基金
中央引导地方科技发展专项资金（桂科ZY22096025）; 国家自然科学基金（No.81560530; 81760589; 81960590; 82273630）; 2023年度中组部“西部之光”访问学者计划; 2023年度广西八桂青年拔尖人才支持计划
