灯盏花素对高糖作用下大鼠腹膜间皮细胞髓样分化因子88、肿瘤坏死因子-α表达的影响

Effects of breviscapine on the expression of MyD88 and TNF-α in rat peritoneal mesothelial cells stimulatedby high glucose

目的:研究灯盏花素对高糖作用下大鼠腹膜间皮细胞(rat peritoneal mesothelial cells,RPMCS)髓样分化因子88(myeloid differentiation factor 88,MyD88)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达的影响。方法:采用腹膜组织块消化法获取RPMCS并进行原代培养,第3代细胞分组培养:(1)对照组;(2)1.5%、2.5%、4.25%葡萄糖组:用1.5%、2.5%、4.25%葡萄糖分别培养细胞4、6、8 h;(3)各浓度组培养细胞6 h后加入灯盏花素干预8 h。MTT法测定各组的OD值,ELISA法检测上清液中MyD88、TNF-α的浓度。结果:腹透液培养后,RPMCS的OD值较对照组显著降低(P<0.01),细胞增殖下降,且葡萄糖浓度越高,刺激时间越长,OD值下降越显著(P<0.05)。加入灯盏花素干预后,各组OD值升高(P<0.05)。各组加入葡萄糖后与对照组比较MyD88、TNF-α表达显著升高(P<0.01),灯盏花素干预后二者的表达下降(P<0.05)。结论:高糖抑制RPMCS增殖,刺激细胞MyD88、TNF-α表达上调,灯盏花素可缓解高糖对细胞增殖的抑制,下调炎症因子的表达,改善腹膜炎症,保护腹膜功能。

Objective To observe the effects of breviscapine on the expression MyD88 and TNF-α in rat peritoneal mesothelial cells （RPMCS） stimulated by high glucose. Methods RPMCS were isolated after tissue enzymatic digested, cultured and identified by phase contrast inverted and immunocytochemistry method. Cells of the third generation were divided into 3 groups= （1） control group; （2） cells grew in glucose concentration of 1.5%, 2.5%, 4.25% for 4, 6 and 8 h; （3） after stimulated by glucose of three different concentration for 6 h, cells were treated with breviscapine for 8 h. The proliferation of RPMCS in each group was examined by MTF colorimeric assay. MyD88, TNF-α in medium of each group were assayed by ELISA. Results The value of OD of RPMCS reduced compared with control group after stimulated by dialysis solution which contained glucose and it was statistically significant （P 〈 0.01）. Such reduction was more manifest with higher concentration glucose and longer treatment time （P 〈 0.05）. When treated with breviscapine, the value of OD of each group increased （P 〈 0.05）. Compared with control group the cells＇ expression of MyD88 and TNF-c~ significantly increased （P 〈 0.01 ）. After treated with breviscapine the expression of the two target value of each group decreased （P 〈 0.05）. Conclusions High concentration glucose inhibits the proliferation of RPMCS in vitro and increases the expression of MyD88 and TNF-c~. Erigeron can alleviate inhibition of the high concentration glucose and decrease the expression of inflammatory factors. It indicated breviscapine could improve peritonitis and protect peritoneal function.