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目的探讨高盐饮食加重脑缺血损伤机制。方法 (1)体内实验:高盐饮食14 d小鼠建立永久性大脑中动脉结扎(pMCAL)模型;2,3,5-三苯基氯化四氮唑(TTC)染色检测小鼠脑组织损伤面积;伊文思蓝染色检测血脑屏障破坏程度;Western blot方法检测小鼠脑组织中微血管内皮细胞紧密连接蛋白claudin-5表达情况。(2)体外实验:Western blot方法检测高盐环境(80 mmol/L)及氧糖剥夺(OGD)+高盐(80 mmol/L)条件下b End.3小鼠脑微血管内皮细胞claudin-5表达情况。结果 (1)TTC染色显示高盐饮食组小鼠脑缺血面积明显高于正常饮食组(P<0.05);伊文思蓝染色显示高盐饮食组小鼠脑内伊文思蓝含量显著高于正常饮食组(P<0.01);Western blot结果显示高盐饮食组小鼠脑组织中claudin-5表达明显低于正常饮食组(P<0.05)。(2)体外实验Western blot结果显示,与正常培养组相比,高盐组脑内微血管内皮细胞claudin-5表达显著降低(P<0.01);与OGD组相比,OGD+高盐组,小鼠脑微血管内皮细胞claudin-5蛋白表达显著降低(P<0.01)。结论高盐饮食加重脑缺血损伤;高盐饮食可下调脑血管内皮细胞紧密连接蛋白claudin-5的表达,破坏血脑屏障,加重脑损伤。
Objective To investigate the mechanism of high salt diet aggravating cerebral ischemia injury. Methods: (1) In vivo experiments: A permanent model of middle cerebral artery occlusion (pMCAL) was established in high-salt diet for 14 days. Brain tissue injury was detected by TTC staining with 2,3,5-triphenyltetrazolium chloride (TTC) Area. Evans blue staining was used to detect the damage of the blood-brain barrier. Western blot was used to detect the expression of claudin-5 in mouse brain microvessels. (2) In vitro experiments: Western blot was used to detect the expression of claudin-5 in brain endothelium of b.3 mouse after high salt (80 mmol / L) and OGD + 80 mmol / L Express the situation. Results (1) TTC staining showed that the area of cerebral ischemia in the high-salt diet group was significantly higher than that in the normal diet group (P <0.05). Evans blue staining showed that the content of Evans blue in the brain of the high-salt diet group was significantly higher than that of the normal diet group (P <0.01). Western blot results showed that the expression of claudin-5 in brain tissue of high-salt diet group was significantly lower than that of normal diet group (P <0.05). (2) Western blot results showed that the expression of claudin-5 in high salt group was significantly lower than that in normal group (P <0.01). Compared with OGD group, OGD + high salt group, mouse The expression of claudin-5 in brain microvascular endothelial cells was significantly decreased (P <0.01). Conclusion High-salt diet aggravates cerebral ischemic injury. High-salt diet can down-regulate the expression of claudin-5, the blood-brain barrier and the brain injury.