Sirt3/Sod2信号通路介导的补骨脂酚抗糖尿病大鼠主动脉凋亡的机制研究

Study on Mechanism of Sirt3/Sod2 Signaling Pathway Mediated Bakuchiol Against Aortic Apoptosis in Diabetic Mellitus Rats

关键词:  糖尿病  补骨脂酚  主动脉  Sirt3/Sod2信号通路  凋亡

Key words:  diabetic mellitus  bakuchiol  aortic  Sirt3/Sod2 signaling pathway  apoptosis

摘要:

目的 探讨补骨脂酚在糖尿病主动脉组织中的保护作用及其机制，明确Sirt3/Sod2信号通路在此过程中的作用。方法 将SD大鼠随机分成对照组、糖尿病组和糖尿病+补骨脂酚组，将内皮细胞分为高脂高糖组、高脂高糖+3-TYP组、高脂高糖+补骨脂酚组和高脂高糖+补骨脂酚+3-TYP组。采用OGTT试验检测大鼠的空腹血糖。采用HE染色观察主动脉壁病理变化。采用荧光染色观察主动脉Sirt3表达变化。采用Western blotting检测主动脉组织及内皮细胞Sirt3、Ac-Sod2、Cleaved caspase-3、Bax、Bcl-2蛋白的表达情况。并用Sirt3的抑制剂3-TYP来研究补骨脂酚抗糖尿病大鼠主动脉凋亡的作用机制。结果 糖尿病大鼠主动脉壁组织排列紊乱，细胞减少，Sirt3及Bcl-2蛋白表达水平明显降低，Ac-sod2、Cleaved caspase-3及Bax蛋白表达水平明显升高。补骨脂酚可显著改善高血糖引起的主动脉壁组织排列紊乱情况，升高Sirt3及Bcl-2蛋白的表达水平，降低Ac-sod2、Cleaved caspase-3及Bax蛋白的表达水平。在高脂高糖损伤内皮细胞试验中Sirt3及Bcl-2蛋白表达水平降低，Ac-sod2、Cleaved caspase-3及Bax蛋白表达升高，而予补骨脂酚处理可升高Sirt3及Bcl-2蛋白的表达水平，降低Ac-sod2、Cleaved caspase-3及Bax蛋白表达水平。予抑制剂3-TYP后，抑制补骨脂酚的保护作用。结论 补骨脂酚共处理可通过激活Sirt3/Sod2信号通路显著抑制糖尿病引起的主动脉壁组织损伤，减少细胞凋亡。

Abstract:

OBJECTIVE To study the protective effect and mechanism of bakuchiol in diabetic aortic artery, and clarify the role of Sirt3/Sod2 signaling pathway in this process. METHODS SD rats were randomly divided into control group, diabetic mellitus group, diabetic mellitus and bakuchiol co-treatment group. Endothelial cells were divided into high fat and high sugar group, high fat and high sugar + 3-TYP group, high fat and high sugar + bakuchiol group, and high fat and high sugar + bakuchiol + 3-TYP group. Fasting blood glucose in rats was detected by OGTT. HE staining was used to observe the pathological changes of aortic vascular wall. The expression of Sirt3 in aortic artery was observed by fluorescence staining. Western blotting was used to detect the expression of Sirt3, Ac-Sod2, Cleaved caspase-3, Bax and Bcl-2 proteins in aortic tissue and endothelial cells. Then the mechanism of bakuchiol against aorta apoptosis in diabetes rats was studied by 3-TYP, an inhibitor of Sirt3. RESULTS In diabetes, the aortic wall tissue was disordered, and the number of cells decreased. The expression of Sirt3 and Bcl-2 protein decreased, while the expression of Ac-sod2, Cleared caspase-3 and Bax protein increased. However, bakuchiol could significantly improve the disorder of aortic wall tissue caused by hyperglycemia. The expression of Sirt3 and Bcl-2 protein increased, while the expression of Ac-sod2, Cleared caspase-3 and Bax protein decreased. After the endothelial cells injured by high fat and high glucose, the expression levels of Sirt3 and Bcl-2 proteins were decreased, and the expression levels of Ac-sod2, Cleaved caspase-3 and Bax proteins were increased. However, bakuchiol could significantly increase the expression levels of Sirt3 and Bcl-2 proteins, and decrease the expression levels of Ac-sod2, Cleaved caspase-3 and Bax proteins. The protective effect of bakuchiol was inhibited by 3-TYP. CONCLUSION Bakuchiol co-treatment could significantly inhibit the aortic artery damage caused by diabetic mellitus and reduced cell apoptosis by activating Sirt3/Sod2 pathway.