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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 change of adhesion function of endothelial cells (VEC) induced by lipopolysaccharide (LPS) and smooth muscle cells (SMC) in rat monocytes and the effect of paeonal (Pae) intervention. Methods: VEC-SMC co-culture model was established by Transwell chamber and VEC-induced injury induced by LPS. MTT assay and LDH assay were used to detect VEC activity ; The levels of IL-1β and TNF-α secreted by VEC were detected by ELISA; the expression of ICAM-1 by immunocytochemistry; the adhesion function of VEC and monocytes were detected by rose bengal method. Results: The concentration of VEC induced by LPS was 100 μg · L-1 for 7 h, and Pae (15,30,60 μmol·L-1) for 24 h inhibited the LPS-induced VEC injury remarkably Increase the LPS-induced decline in VEC survival rate; reduce VEC secretion of IL-1β and TNF-α levels, while reducing the expression of ICAM-1; thereby inhibiting LPS-induced VEC and monocyte adhesion. CONCLUSION: Pae can protect VEC from LPS injury by inhibiting the expression of IL-1β and TNF-α, and can inhibit the adhesion of VEC to monocytes by decreasing the expression of ICAM-1.