LPS刺激对大鼠视网膜小胶质细胞CD80、CD86和CD40表达的影响

Effects of LPS stimulation on expression of CD80,CD86 and CD40 in rat retinal microglial cells

目的研究脂多糖(LPS)刺激对CD80、CD86、CD40在大鼠视网膜小胶质细胞上表达的影响,探讨视网膜小胶质细胞在视网膜和视神经病变中与T细胞活化的关系。方法分离培养大鼠视网膜小胶质细胞并进行OX42染色鉴定,当细胞培养达80%融合状态时分为正常培养组和LPS刺激组。采用RT-PCR法检测两组细胞CD80、CD86和CD40 mRNA的表达,硝酸还原法和ELISA法检测两组细胞一氧化氮(NO)和肿瘤坏死因子-α(TNFα-)的分泌量。结果RT-PCR检测显示,LPS刺激组的视网膜小胶质细胞CD80、CD86和CD40 mRNA的表达较正常培养组显著增加,两组比较差异有统计学意义(均P<0.01)。正常培养组的视网膜小胶质细胞有少量NO和TNFα-分泌,LPS刺激组的小胶质细胞培养上清液中NO和TNFα-的分泌量较正常组显著增加[(18.10±1.57)μmol/Lvs(1.55±0.15)μmol/L,(51.07±4.30)pg/mLvs(2.37±0.31)pg/mL],两组比较差异有统计学意义(均P<0.01)。结论培养的大鼠视网膜小胶质细胞有共刺激分子表达,LPS刺激后表达上调,提示视网膜小胶质细胞可能与T细胞活化有关。

Objective To study the effects of LPS stimulation on the expression of CD80, CD86 and CD40 in rat retinal microglial cells, and investigate the relationship between microglial cells and T cell activation in the retinopathy and optic neuropathy. Methods After isolation and cultivation, the rat retinal microglial cells were identified by OX42 staining. Cells were divided into normal culture group and LPS stimulation group when cell merger accounted for 80%. The expression of CD80, CD86 and CD40 mRNA of retinal microglial cells was detected by RT-PCR before and after LPS stimulation. The production of nitric oxide （NO） and tumor necrosis factor （TNF） -α of retinal microglial cells was measured by nitric acid reduction and ELISA, respectively. Results The expression of CD80, CD86 and CD40 mRNA of retinal mieroglial cells was significantly higher in LPS stimulation group than that in normal culture group （P 〈0.01）. The production of NO and TNF-α in the supernatant of retinal microglial cells was significantly higher in LPS stimulation group than that in normal culture group [ （ 18. 10 ＋ 1.57） μmol/L vs（ 1.55 ± 0.15） μmol/L and （51.07 ± 4.30） pg/mL vs（2.37 ± 0.31） pg/mL, respectively] （P 〈 0.01）. Conclusion The cultivated retinal microglial cells express eostimulatory molecules, and the expression increases after stimulation by LPS, indicating that the retinal microglial cells may be related to T cell activation.