脑复聪对脂多糖激活的小胶质细胞合成环氧合酶-2和分泌前列腺素E2作用的实验研究

摘要

目的 观察中药脑复聪(NFC)含药血清对脂多糖(LPS)激活后的小鼠神经小胶质细胞BV-2的形态、BV-2中环氧合酶-2(COX-2)蛋白的表达以及BV-2上清液中前列腺素E2(PGE2)分泌量水平的影响.方法 将体外培养的小鼠神经小胶质细胞BV-2分成:正常对照组、LPS组、NFC血清组(高、中、低剂量)培养24h,用倒置相差显微镜观察细胞形态,蛋白印迹方法检测BV-2中COX-2蛋白的表达,酶联免疫吸附法测定培养BV-2上清液中PGE2蛋白的浓度.结果 (1)NFC各组对LPS所致的BV-2细胞形态变化均有一定的恢复作用.(2)LPS组BV-2中COX-2含量较正常对照组升高；与LPS组相比,NFC含药血清各剂量组COX-2蛋白含量均下降.(3)BV-2上清液中PGE2蛋白的浓度LPS组较正常对照组含量升高；与LPS组相比,NFC各剂量组PGE2含量均下降.结论 NFC含药血清可以显著下调LPS激活后BV-2中COX-2蛋白的表达量并显著减少BV-2上清液中PGE2的分泌量,减轻炎症反应,改善细胞形态.%Objective To observe the effects of Naofucong serum on morphology of the mouse microglia BV-2,expression of COX-2 protein in BV-2,and the level of PGE2 secreted in the supernatant of the BV-2 culture after lipopolysaccharide activation.Methods Mouse microglia BV-2 cells incubated in vitro were divided into three groups:the normal control group,the LPS group and the high-,moderate-and low-dose of NFC serum groups.After 24h incubation,the morphology of microglia was observed with inverted phase contrast microscope,and the expression of the COX-2 protein was detected by Western blot,and the level of PGE2 protein in the supernatant of BV-2 culture was determined by enzyme-linked immunosorbent assay.Results (1) Each concentration of NFC-containing serum group had a certain restoring effect on the LPS-induced morphological changes.(2) The level of COX-2 protein in BV-2 in the LPS group was significantly increased compared to the control group ; while the levels of COX-2 protein in BV-2 in all of the various doses of NFC-containing serum groups were decreased compared to the LPS group.(3)The culture supernatant of BV-2 in the LPS group exhibited significantly increased level of PGE2 compared to the normal control group ; while all of the various doses of NFC-containing serum groups exhibited decreased PGE2 level compared to the LPS group.Conclusions The NFC-containing serum is able to significantly down regulate the expression of COX-2 protein in BV-2 activated by LPS and reduce the secretion of PGE2 in the supernatant of BV-2 culture.Inflammation reaction can be relieved while morphology of cells can be improved.