目的观察丹皮酚(Paeonal,Pae)对高脂血清诱导的大鼠单核细胞(mononuclear cell,MC)与大鼠主动脉内皮细胞(rat aortic endothelial cells,RAECs)黏附的影响及对RAECs损伤的保护作用。方法高脂乳剂灌胃大鼠并制备高脂血清;组织块预消化贴壁法培养RAECs;淋巴细胞分离液分离MC;孟加拉玫瑰红活细胞染色法测定MC与RAECs的黏附功能;甲基噻唑基四唑比色法以及台盼蓝死细胞染色法检测Pae对高脂血清诱导的RAECs损伤的保护作用。结果30~70ml/L高脂血清呈剂量依赖性地诱导MC与RAECs的黏附,50ml/L时促进MC与RAECs黏附达最大值;高脂血清与RAECs共育6h即可明显促进黏附作用,24h时达最大诱导效应。Pae在浓度为15~240μmol/L时,呈剂量依赖性地抑制高脂血清诱导的黏附作用;240μmol/L孵育24h时抑制作用最明显;Pae在30,60,120μmol/L浓度时,对高脂血清损伤的大鼠RAECs有明显的保护作用。结论Pae对高脂血清诱导的大鼠MC与RAECs黏附功能有抑制效应,对高脂血清损伤的大鼠RAECs有明显的保护作用。
Objective To investigate the effect of Paenonal (Pae) on the adhesion reaction of mononuclear cell (MC) with rat aortic endothelial cells (RAECs) induced by hyperlipidemia serum and the protection of Pae against the injury of RAECs. Methods The rat hyperlipidemia serum was prepared using intragastric injection of high fat emulsion. RAECs were obtained using method of predigestion and tissue pieces adherence. MC was isolated using lymphocyte separation medium. The adhesion reaction of MC with RAECs induced by hyperlipidemia serum was detected by Rose Bengal staining. The protection of Pae against hyperlipidemia serum induced injury of RAECs was assayed using methyl thiazolyl tetrazolium staining and trypan blue staining. Results Hyperlipidemia serum 30-70 ml/L induced the adhesion reaction of MC with RAECs in a dose-dependent manner, and the adhesion reaction reached to the maximum at the concentration of 50 ml/L. Six hours after co-culture of hyperlipidemia serum with RAECs, an obvious adhesion reaction could be observed; and the adhesion reaction reached to the maximum after 24 hours of coculture. Pae 15-240μmol/L inhibited the adhesion reaction induced by hyperlipidemia serum in a dose-dependent manner; And the inhibitive effect of Pae reached to the maximum at the concentration of 240 μmol/L after 24 hours of co-culture. Pae 30, 60 and 120μmol/L has marked protection against the injury of RAECs induced by hyperlipidemia serum, respectively. Conclusion Pae could inhibit the adhesion reaction of MC with RAECs and obviously protect RAECs from injury induced by hyperlipidemia serum.