血必净注射液通过降低细胞紧密连接的通透性减轻LPS诱导的脓毒症大鼠肺微血管内皮细胞损伤

Xuebijing injection attenuated LPS-induced pulmonary microvascular endothelial cell injury in septic rats by reducing the permeability of tight junction

目的探究血必净注射液(XBJ)对脂多糖诱导的脓毒症大鼠肺微血管内皮细胞损伤的作用及其对细胞紧密连接通透性的影响。方法选择大鼠肺微血管内皮细胞,利用5μg/mL的脂多糖(LPS)处理大鼠肺微血管内皮细胞24 h,构建大鼠脓毒症肺损伤细胞模型,XBJ治疗组采用生药浓度2.5、5、10、25、50 mg/mL的XBJ共同进行干预24 h。CCK-8检测各组细胞活性,筛选最佳XBJ浓度;Hoechst染色检测各组细胞凋亡率;Western blot检测各组细胞凋亡相关蛋白Bax、Bcl-2和cleaved caspase3的表达;Transwell小室检测单层大鼠肺微血管内皮细胞通透性;Western blot检测紧密连接相关蛋白ZO-1、ZO-2和occludin的表达。结果CCK8实验结果显示,XBJ的最佳浓度为10mg/mL。LPS处理后,大鼠细胞凋亡率明显升高,凋亡相关蛋白Bax/Bcl-2的比值和cleaved caspase3的表达水平增加;而XBJ治疗后,细胞凋亡率下降。LPS处理导致FITC-dextran含量显著增加,ZO-1、ZO-2和Occludin的蛋白表达水平降低;而XBJ治疗抑制LPS引起的细胞通透性增加,改善LPS对细胞紧密连接的抑制。结论血必净注射液通过降低细胞紧密连接的通透性减轻LPS诱导的脓毒症大鼠肺微血管内皮细胞损伤。

Objective To investigate the effect of Xuebijing injection(XBJ)on lipopolysaccharide-induced injury of pulmonary microvascular endothelial cells in septic rats and its effect on the permeability of tight junctions.Methods Pulmonary microvascular endothelial cells were selected,and cells were treated with 5μg/mL LPS for 24 h to establish a cell model of sepsis-induced lung injury.XBJ treatment group was treated with XBJ at the concentration of 2.5,5,10,25,and 50 mg/mL for 24 h.CCK-8 method was used to detect the cell activity and the optimal concentration of XBJ was screened.Hoechst staining was used to detect the apoptosis rate of cells.Western blot method was used to detect the expressions of apoptosis-related proteins Bax,Bcl-2,and cleared caspase3.Transwell method was used to detect the permeability of pulmonary microvascular endothelial cells.Western blot was used to detect the expressions of tight junction related proteins ZO-1,ZO-2,andoccludin.Results The results of CCK8 showed that the optimal concentration of XBJ was 10 mg/mL.After LPS treatment,the apoptosis rate of cells was increased significantly,and the ratio of Bax/Bcl-2 and the expression of cleaved caspase3 were increased.After XBJ treatment,the apoptosis rate of cells was decreased.LPS treatment significantly increased the content of FITC-dextran,and decreased the expressions of ZO-1,ZO-2 and Occludin.XBJ treatment inhibited the increase of cell permeability induced by LPS and improved the inhibition of tight cell connection by LPS.Conclusion Xuebijing injection can alleviate LPS induced pulmonary microvascular endothelial cell injury in septic rats by reducing the permeability of tight junctions.