丹皮酚对脂多糖诱导损伤的与平滑肌细胞共培养的大鼠血管内皮细胞黏附功能的影响

Effect of paeonol on adhesive function of rat vascular endothelial cells induced by lipopolysaccharide and co-cultured with smooth muscle cells

目的:观察脂多糖(LPS)诱导的与平滑肌细胞(SMC)共培养的内皮细胞(VEC)与大鼠单核细胞黏附功能的改变及丹皮酚(paeonal,Pae)的干预作用。方法:组织块预消化贴壁法原代培养大鼠血管内皮细胞和大鼠血管平滑肌细胞;采用Transwell小室建立VEC-SMC共培养模型;采用LPS诱导VEC的损伤;MTT法和LDH法检测VEC活力;ELISA法检测VEC分泌IL-1β和TNF-α的水平;免疫细胞化学法检测ICAM-1表达水平;孟加拉玫瑰红染色法检测VEC与单核细胞的黏附功能。结果:LPS诱导VEC损伤的质量浓度为100μg·L-1,时间为7 h;Pae(15,30,60μmol·L-1)对以上细胞模型作用24 h可有效抑制LPS诱导的VEC损伤,显著提高LPS导致的VEC存活率的下降;降低损伤VEC分泌IL-1β和TNF-α的水平,同时减少ICAM-1的表达;从而抑制LPS诱导的VEC与单核细胞的黏附。结论:Pae通过抑制IL-1β和TNF-α表达而保护VEC免受LPS的损伤,可以通过降低ICAM-1的表达水平达到抑制VEC与单核细胞的黏附的作用。

Objective： To observe the changes in the adhesive function of vascular endothelial cells （VEC） and rat monocytes induced by lipopolysaccharide （LPS） and co-cultured with smooth muscle cells （SMC） and the intervention effect of paeonol （Pae）. Method： Primary rat vascular endothelial cells （VECs） and rat vascular smooth muscle cells （VSMCs） were cultured by predigesting and adhering tissue blocks. The VEC-VSMC co-culture model was established by Transwell chamber. LPS was used to induce VEC injury. MTT assay and LDH assay were used to determine the VEC activity. ELISA assay was used to detect IL-1β and TNF-α secreted by the VEC. The immunocytochemistry assay was carried out to detect the expression of ICAM-1. The Rose Bengal Staining was used to test adhesive function between VECs and monocytes. Result： The concentration of LPS-induced VEC injury was 100 μg·L^-1, and the time was 7 h. after the intervention on the above cell model for 24 h, Paeonol （15, 30, 60 μmol·L^-1） could effectively inhibit LPS-induced VEC injury and VEC injury, significantly enhance the survival rate of LPS-injured VECs, decrease IL-1β and TNF-α secreted by the injured VEC, and reduce the expression of ICAM-1, so as to inhibit the adhesion of LPS-induced VECs and monocytes. Conclusion： Paeonol could inhibit IL-1β and TNF-α expression to protect VECs from being injured by LPS, and reduce ICAM-1 expression to inhibit the adhesion between VECs and monocytes.