Department of Pediatrics, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Eur Rev Med Pharmacol Sci. 2018 Nov;22(22):7994-8003. doi: 10.26355/eurrev_201811_16428.
To discuss the effect of complement C52 (C5a) and complement C5a receptor (C5aR) antagonists on inflammatory status of mouse microglial cells.
Primary culture was performed on mouse microglial cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to detect effect of C5a and C5aR antagonists on vitality of microglial cells. The effect of C5a and C5aR antagonists on mRNA expression of p38MAPK and ERK1/2 was determined using quantitative PCR (qPCR). Enzyme linked immunosorbent assay (ELISA) was used to measure expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cells.
According to quantitative fluorescent PCR, relative expression of p38MAPK and ERK1/2 mRNA in C5a antagonist treatment group was significantly higher compared to normal group and C5a+C5aR antagonist treatment group (p<0.05). However, the relative expression of the C5a+C5aR antagonist treatment group was significantly lower compared to that of the normal group (p<0.05). Expression of Iba1, p-p38MAPK and p-ERK1/2 proteins in C5a antagonist treatment group was significantly higher than normal group, in C5a+C5aR antagonist treatment group was lower than C5a antagonist treatment group (p<0.05). There were significant differences for IL-6 and TNF-α levels among 5 groups (p<0.05). Expression of cytokines was the highest in 100 nM C5a antagonist treatments and lowest in normal group.
Complement C5a upregulated expression of inflammatory factors in mouse microglial cells, while C5aR antagonist inhibited occurrence and progression of inflammatory status. This was achieved by affecting transcriptional and translational processes of different factors in p38MAPK and ERK1/2 signaling pathway.
探讨补体 C52（C5a）和补体 C5a 受体（C5aR）拮抗剂对小鼠小胶质细胞炎症状态的影响。
对小鼠小胶质细胞进行原代培养。噻唑蓝（MTT）比色法检测 C5a 和 C5aR 拮抗剂对小胶质细胞活力的影响。采用实时荧光定量 PCR（qPCR）检测 C5a 和 C5aR 拮抗剂对 p38MAPK 和 ERK1/2 mRNA 表达的影响。酶联免疫吸附试验（ELISA）检测细胞中白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的表达。
根据定量荧光 PCR，C5a 拮抗剂治疗组 p38MAPK 和 ERK1/2 mRNA 的相对表达明显高于正常组和 C5a+C5aR 拮抗剂治疗组（p<0.05）。然而，C5a+C5aR 拮抗剂治疗组的相对表达明显低于正常组（p<0.05）。C5a 拮抗剂治疗组 Iba1、p-p38MAPK 和 p-ERK1/2 蛋白的表达明显高于正常组，C5a+C5aR 拮抗剂治疗组低于 C5a 拮抗剂治疗组（p<0.05）。5 组间 IL-6 和 TNF-α 水平差异有统计学意义（p<0.05）。100 nM C5a 拮抗剂处理组细胞因子表达最高，正常组最低。
补体 C5a 上调了小鼠小胶质细胞炎症因子的表达，而 C5aR 拮抗剂抑制了炎症状态的发生和发展。这是通过影响 p38MAPK 和 ERK1/2 信号通路中不同因子的转录和翻译过程来实现的。
