IL-1β诱导体外培养大鼠脑微血管内皮细胞的损伤

IL-1β induced damage of rat brain microvascular endothelial cells in vitro

目的 探讨IL-1β对大鼠脑微血管内皮细胞的结构和功能的损伤作用。方法 体外培养大鼠来源的脑微血管内皮细胞,将实验分为4组:Control组、IL-1β组、IL-1β+IL-1RA组、IL-1RA组。用TEER和FITC-Dextran检测各组细胞的渗透性,免疫荧光法和Western blot方法检测Claudin-5、Occludin、ZO-1的表达变化,用鬼笔环肽(TRITC Phalloidin)检测内皮细胞F-actin的改变。结果 (1)IL-1β组的TEER显著低于Control组,而与IL-1β组相比,IL-1β+IL-1RA组TEER值又显著升高;(2)IL-1β组FITC-Dextran的渗漏含量显著高于Control组,而与IL-1β组相比,IL-1β+IL-1RA组的FITC-Dextran渗漏含量显著降低;(3)与Control组相比,IL-1β组内Claudin-5、Occludin、ZO-1的表达水平均显著降低,而与IL-1β组相比,IL-1β+IL-1RA组Claudin-5、Occludin、ZO-1的表达均明显升高;(4)与Control组相比,IL-1β组F-actin蛋白结构破坏较为明显。结论 IL-1β诱导了大鼠脑微血管内皮细胞结构和功能的损伤。

Objective To investigate the effects of IL-1β on the structure and function of rat brain microvascular endothelial cells. Methods Rat brain microvascular endothelial cells were cultured in vitro, and the experiments were divided into four groups: Control, IL-1β, IL-1β+IL-1RA and IL-1RA. The permeability of monolayer rat brain microvascular endothelial cells was measured by TEER and FITC-Dextran methods in each group. Immunofluorescence staining and Western Blot were used to detect the morphological changes of tight junction proteins Claudin-5, Occludin and ZO-1. F-actin was stained with TRITC Phalloidin to observe its destruction. Results(1) The TEER of IL-1β group is significantly lower than that of the control group, while the TEER of IL-1β+IL-1RA group has increased more significantly than that of IL-1β group.(2) The FITC-Dextran leakage of IL-1β group increased more significantly than that of the control group, while the FITC-Dextran infiltration of IL-1β+IL-1RA group has decreased more significantly than the that of IL-1β group.(3) The expression of Claudin-5, Occludin and ZO-1from IL-1β group has decreased more significantly than that of the control group, while the expressions of Claudin-5, Occludin and ZO-1 from IL-1β+IL-1RA group has increased more significantly, compared to IL-1β group.(4) The structure of F-actin was destroyed significantly in IL-1β group when compared to the control group. Conclusion IL-1β induced structural and functional damage of microvascular endothelial cells of rat brain.