6mmol/L葡萄糖和24.4mmol/L甘露醇的DMEM/F12培养液。各组培养48h用MTT法检测各组视网膜色素上皮细胞的增生活力,用CM-H2DCFDA荧光染色检测RPE细胞中ROS的产生量。结果:与对照组相比,高糖培养人视网膜色素上皮细胞48h可以导致RPE细胞的损伤,抑制RPE细胞增殖,并使ROS生成增加,在一定程度上,与p38-MAPK途径的激活有关。结论:高糖培养ARPE-19可致细胞损伤,抑制增殖,并使ROS生成增加。
The tissue destruction characteristic of syphilis infection may be caused by inflammation due to Treponema pallidum and the ensuing immune responses to the pathogen.T.pallidum 许多 membrane proteins are thought to be potent inducers
of inflammation during the early stages of infection.However,the actual membrane proteins that induce inflammatory cytokine production are not known,nor are the molecular mechanisms responsible for triggering and sustaining the inflammatory cascades.In the present study,Tp0751 recombinant protein from T.pallidum was found to induce the production of proinflammatory cytokines,including TNF-α,IL-1βand
IL-6,in a THP-1 human monocyte cell line.The signal transduction pathways involved in the production of these cytokines were 确认细节 then further investigated.No inhibition of TNF-a,IL-1β,or IL-6 production was observed following treatment with the SAPK/JNK specific inhibitor SP600125 or with an 17-AAG核磁共振 ERK inhibitor PD98059.By contrast,anti-TLR2 mAb,anti-CD14 mAb,and the p38 inhibitor SB203580 significantly inhibited the production of all three cytokines.In addition,pyrrolidine dithiocarbamate (PDTC),a specific inhibitor of NF-κB,profoundly inhibited the production of these cytokines.Tp0751
treatment strongly activated NF-κB,as revealed by Western blotting.However,NF-κB translocation was significantly inhibited by treatment with PDTC.These results indicated that TLR2,CD14,MAPKs/p38,and NF-κB might be implicated in the inflammatory reaction caused by T.pallidum infection.
目的:研究脂多糖(LPS)对肥大细胞高迁移率族蛋白B1(HMGB1)释放的诱导作用及其胞内信号通路。方法:采用小鼠肥大细胞株P815,观察不同剂量LPS诱导后细胞培养上清液中HMGB1含量和细胞HMGB1mRNA表达水平的变化,及不同剂量丝裂原活化蛋白激酶(MAPK)信号通路抑制剂(SB203580、SB202190、U0126和PD98059)对LPS诱导后HMGB1胞外释放的影响。HMGB1含量采用酶联免疫吸附试验检测。结果:LPS诱导后细胞培养上清液中HMGB1含量明显升高,并与LPS剂量、诱导时间有关;100μg/LLPS分别诱导8、24、48h后,HMGB1mRNA表达水平明显增强(P<0.01);SB203580和SB202190对LPS诱导HMGB1释放有明显的抑制作用(P<0.