川芎嗪对脂多糖诱导的内皮细胞损伤的保护作用

Protective effect of tetramethylpyrazine on the injury endothelial cell induced by lipopolysaccharide

目的：探讨川芎嗪（tetramethylpyrazine,TMP）对脂多糖（lipopolysaccharide,LPS）诱导的人脐静脉内皮细胞（human umbilical vein endothelial cell,HUVEC）Rho A、Rho相关卷曲螺旋形成蛋白激酶2（Rho associated coiled coil forming protein kinase,ROCK2）、肌球蛋白轻链（myosin light chain,MLC）磷酸化及细胞骨架连接蛋白（ezrin-radxin-moesin,ERM）磷酸化水平的影响。方法：以体外培养的HUVEC为实验对象,分为对照组、LPS组和TMP组（0.5、1.0、1.5 mg/m L）,荧光定量PCR测定内皮细胞Rho A、ROCK2和信使RNA（p-Ezr mRNA）的表达,Western blot法测定内皮细胞Rho A、ROCK2、p-MLC和p-ERM蛋白水平的表达。结果：与对照组比较,LPS组Rho A、ROCK2、p-MLC和p-ERM蛋白及Rho A、ROCK2和p-Ezr mRNA表达均显著升高（P〈0.01）;与LPS组比较,TMP 3组Rho A蛋白表达与mRNA表达差异均无统计学意义（P〉0.05）,ROCK2、p-MLC和p-ERM蛋白及ROCK2、p-Ezr mRNA表达均降低（P〈0.05～P〈0.01）,且TMP 3组之间ROCK2、p-MLC和p-ERM的分子表达差异均有统计学意义（P〈0.05～P〈0.01）。结论：TMP对LPS诱导的HUVEC损伤的保护作用可能是通过抑制Rho/ROCK信号通路中ROCK2、p-MLC和p-ERM的表达,以减弱LPS对内皮细胞骨架的损伤。

Objective： To explore the effects of tetramethylpyrazine（ TMP） on the levels of Rho A,Rho associated coiled coil forming protein kinase 2（ ROCK2）,myosin light chain（ MLC） phosphorylation and ezrin-radxin-moesin（ ERM） phosphorylation in the injury of human umbilical vein endothelial cells（ HUVECs） induced by lipopolysaccharide（ LPS）. Methods： The HUVECs were cultured in vitro,and divided into the control group,LPS group and TMP groups（ 0. 5,1. 0,1. 5 mg / m L）. The mRNA levels of Rho A,ROCK2 and p-EZR in endothelial cells were detected by real-time fluorescence quantitative PCR,and the protein levels of Rho A,ROCK2,p-MLC and p-ERM were examined using western blotting. Results： Compared with the control group,the protein levels of Rho A,ROCK2,p-MLC and p-ERM,and the mRNA levels of Rho A,ROCK2 and p-Ezr in LPS group significantly increased（ P〈0. 01）. The differences of the protein and mRNA levels of Rho A between LPS group and 3 TMP groups were not statistically significant（ P〈0. 05）. Compared with the LPS group,the protein levels of ROCK2,p-MLC and p-ERM,and the mRNA levels of ROCK2 and p-Ezr decreased significantly in 3 TMP groups（ P〈0. 01）. The differences of the expression of ROCK2,p-MLC and p-ERM within 3 TMP groups were statistically significant（ P〈0. 05 to P〈0. 01）. Conclusions： TMP may protect the HUVECs against LPS-induced injury by inhibiting the expressions of ROCK2,p-MLC and p-ERM in Rho / ROCK signaling pathways,and reducing the injury of LPS on endothelial cell skeleton.