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目的:研究丹参酚酸B对脑缺血/再灌注(Cerebral ischemia/reperfusion,CI/R)损伤的保护作用及机制。方法:通过结扎颈总动脉缺血2 h再灌注48 h复制CI/R模型,将实验大鼠随机分为假手术组、模型组、丹参酚酸B组,每组10只,培养大脑皮层神经细胞,分别给予0,10,25,50 umol/L的丹参酚酸B。通过2,3,5-氯化三苯基四氮唑蓝(TTC)染法测定大鼠脑梗死面积,Western Blot检测大鼠Nrf2和HO-1蛋白表达水平以及细胞中Nrf2和HO-1蛋白表达水平。再通过细胞缺氧缺糖模型,检测不同浓度丹参酚酸B对于细胞死亡率及细胞内ROS水平以及转染Nrf2或HO-1 si RNA后细胞死亡率及细胞内ROS水平。结果:与模型组比较,丹参酚酸B组的大鼠脑梗死面积明显减小,脑组织中Nrf2和HO-1蛋白表达水平均明显增加(P<0.05)。大脑皮层细胞中,随着丹参酚酸B浓度增加,细胞HO-1蛋白及细胞核中Nrf2蛋白表达水平逐渐提高,而细胞质中Nrf2蛋白表达水平逐渐降低(P<0.05)。细胞缺糖缺氧条件下,与对照组相比,丹参酚酸B组均能够降低细胞的死亡率及细胞内ROS水平,敲除Nrf2或HO-1后,丹参酚酸B组的细胞死亡率与细胞内ROS水平均有明显减低(P<0.05)。结论:丹参酚酸B对大鼠CI/R具有保护作用,其作用机制可能通过Nrf2/HO-1减轻CI/R所造成的氧化应激损伤。
Objective: To study the protective effect of salvianolic acid B on cerebral ischemia / reperfusion (CI / R) injury and its mechanism. Methods: CI / R model was established by ligating the common carotid arteries 2 hours after reperfusion and reperfused for 48 hours. The rats were randomly divided into sham operation group, model group and salvianolate group B, with 10 rats in each group. Cells were given 0,10,25,50 umol / L of salvianolic acid B. The area of cerebral infarction in rats was determined by TTC staining with 2,3,5,5-trichlorotoluene. The expression of Nrf2 and HO-1 protein and the expression of Nrf2 and HO-1 protein The expression level. The cell death rate, intracellular ROS level and the cell death rate and intracellular ROS level after transfection of Nrf2 or HO-1 si RNA were detected by cell hypoxia-glucose deprivation model. Results: Compared with the model group, the area of cerebral infarction in Salvia miltiorrhiza B group was significantly reduced, and the expression of Nrf2 and HO-1 protein in brain tissue were significantly increased (P <0.05). With the increase of salvianolic acid B concentration, the level of Nrf2 protein in the cell HO-1 protein and nucleus gradually increased, while the level of Nrf2 protein in the cytoplasm decreased gradually (P <0.05). Under hypoxia and hypoxia conditions, compared with the control group, Salvia miltiorrhiza B group were able to reduce the cell death rate and intracellular ROS levels, the cell death rate of Salvia miltiorrhiza B group after knocking out Nrf2 or HO-1 And intracellular ROS levels were significantly reduced (P <0.05). CONCLUSION: Salvia miltiorrhiza B is protective against CI / R in rats, and its mechanism may be related to the reduction of oxidative stress induced by CI / R via Nrf2 / HO-1.