Tubeimoside I通过激活SIRT3改善脑缺血/再灌注损伤

Shaoyue Huang, Zhen Hong, Kuo Li
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摘要

脑缺血再灌注(Cerebral ischemic -reperfusion, CIR)是一种伴有脑组织炎症和细胞凋亡的疾病,严重影响人类健康和生命。Tubeimoside I (TBMS-1)能抑制神经炎症,具有神经保护作用;然而,其对脑缺血再灌注(IR)损伤的影响尚不清楚。采用小鼠脑动脉闭塞/再灌注模型模拟CIR损伤。采用2,3,5-三苯四唑氯染色法观察大鼠神经功能及脑梗死面积。采用酶联免疫吸附血清学检测试剂盒检测肿瘤坏死因子-α、白细胞介素(IL)-1β、IL-6和IL-10水平。蛋白印迹法检测细胞凋亡相关因子的表达。此外,采用氧-葡萄糖剥夺/再氧化(OGD/R)处理PC12(嗜铬细胞瘤)细胞,建立体外CIR损伤模型。采用细胞计数试剂盒-8检测细胞活力,流式细胞术检测细胞凋亡水平。体内实验结果显示,Tubeimoside I可减小脑梗死面积,降低炎症因子含量,抑制凋亡相关因子Bax、裂解caspase-3 (Asp175)的表达,促进b细胞淋巴瘤蛋白2等存活因子的表达。在体外,Tubeimoside I能够提高细胞活力,抑制细胞凋亡。从机制上讲,Tubeimoside I能够增强nad依赖性去乙酰化酶sirtuin-3 (SIRT3)在体内和体外的表达。SIRT3抑制剂可消除Tubeimoside I对OGD/ r处理细胞的保护作用。Tubeimoside I通过激活SIRT3减轻CIR损伤。因此,它可能是治疗脑IR损伤的潜在候选药物。
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Tubeimoside I ameliorates cerebral ischemia/reperfusion injury through activating SIRT3
Cerebral ischemia-reperfusion (CIR) seriously affects human health and life as it is accompanied by inflammation and apoptosis in brain tissues. Tubeimoside I (TBMS-1) can inhibit neuroinflammation and has neuroprotective effects; however, its effects on ischemia-reperfusion (IR) injury of the brain requires clarity. A mouse cerebral artery occlusion/reperfusion model was used to simulate CIR injury. The neurological function and the area of cerebral infarction were assessed by 2,3,5-triphenyltetrazolium chloride staining. Tumor necrosis factor-α, Interleukin (IL)-1β, IL-6, and IL-10 levels were measured by enzyme-linked-immunosorbent serologic assay kits. Protein blot analysis was performed to assess the expression of apoptosis-related factors. In addition, PC12 (pheo-chromocytoma) cells were treated with oxygen–glucose deprivation/reoxygenation (OGD/R) to establish an in vitro model of CIR injury. The cell viability was measured by cell counting kit-8 assay, and apoptosis levels were detected by flow cytometry. In vivo results indicated that Tubeimoside I reduced cerebral infarct size, decreased inflammatory factor content, inhibited the expression of apoptosis-related factors, including Bax and cleaved-caspase-3 (Asp175), and promoted the expression of survival factor, such as B-cell lymphoma protein 2. In vitro, Tubeimoside I was able to increase cell viability and inhibit apoptosis. Mechanistically, Tubeimoside I was able to enhance both in vivo and in vitro expressions of NAD-dependent deacetylase sirtuin-3 (SIRT3). SIRT3 inhibitor abolished the protective effect of Tubeimoside I on OGD/R-treated cells. Tubeimoside I lessened CIR injury by activating SIRT3. Hence, it could be a potential drug candidate for treating IR injury of the brain.
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