法舒地尔通过Rho-ROCK通路抑制高糖诱导的单核细胞与内皮细胞黏附

目的观察Rho激酶抑制剂法舒地尔能否抑制高糖诱导的单核细胞(THP-1)与人脐静脉内皮细胞黏附,初步探讨其潜在机制。方法使用荧光显微镜观察荧光标记的THP-1与人脐静脉内皮细胞黏附。血管细胞黏附分子1和单核细胞趋化蛋白1的mRNA及蛋白表达水平分别通过RT-PCR、Western blot检测。使用Western blot检测RhoA、ROCK-1、p-MYPT及MYPT蛋白表达水平变化。结果法舒地尔显著抑制高糖诱导的THP-1与人脐静脉内皮细胞黏附,并呈剂量依赖性,其中,低浓度法舒地尔(10-6 mmol/L)干预24 h黏附减少约33.4%,高浓度法舒地尔(10-5 mmol/L)干预24 h黏附减少约42.8%(P值均0.05),干预12 h组趋势相同。法舒地尔显著减低高糖诱导的人脐静脉内皮细胞血管细胞黏附分子1及单核细胞趋化蛋白1的mRNA及蛋白表达水平,并呈时间依赖性和剂量依赖性。高糖显著增加p-MYPT/MYPT比值,而法舒地尔减弱高糖对p-MYPT/MYPT比值的影响,抑制Rho/RCOK通路激活。结论法舒地尔抑制高糖诱导的THP-1与人脐静脉内皮细胞黏附,减低高糖诱导的人脐静脉内皮细胞血管细胞黏附分子1和单核细胞趋化蛋白1的表达,减少高糖诱导的Rho/RCOK通路激活,提示法舒地尔可能成为新的糖尿病大血管病变治疗药物。

Fasudil Inhibiting High Glucose Induced Monocyte-Endothelial Cells Adhesion Through Rho/ROCK Pathway

Aim To investigate whether Rho kinase inhibitor fasudil inhibits high glucose(HG)-induced cell adhesion to human umbilical vein endothelial cells(HUVEC) and search for its underlying mechanisms. Methods The adhesion of monocytes to HUVEC was determined using fluorescence-labeled monocytes.The mRNA and protein levels of vascular cell adhesion molecule-1(VCAM-1) and monocyte chemotactic protein 1(MCP-1) were measured using real-time polymerase chain reaction(RT-PCR) and Western blot.The amounts of RhoA,ROCK1,p-MYPT and MYPT were determined using Western blot analysis. Results Fasudil significantly suppressed HG-induced adhesion of THP-1 to HUVEC in a dose manner: namely by about 33.4% in the presence of low dose(10-6 mmol/L) and by 42.8% in high dose(10-5 mmol/L) at 24 hours(both P<0.05);the same for 12 h.Fasudil significantly suppressed the HG-induced increase of mRNA and protein expression of VCAM-1 and MCP-1 in dose and time manner.HG markedly increased ratio of p-MYPT/MYPT,while fasudil inhibited HG-induced activation of Rho/ROCK pathway. Conclusion The adhesion of monocytes to HUVEC and the expression of VCAM-1and MCP-1 induced by high glucose could be inhibited by fasudil,the same to activation of Rho/ROCK pathway.It suggests that fasudil may represent a new treatment for diabetic vascular injury.