微生物与感染
微生物與感染
미생물여감염
JOURNAL OF MICROBES AND INFECTION
2015年
3期
159-165
,共7页
秦园%曹浚垣%吴淑燕%黄瑞
秦園%曹浚垣%吳淑燕%黃瑞
진완%조준원%오숙연%황서
脂多糖%巨噬细胞%CD36%胞外信号调节激酶通路%炎症因子
脂多糖%巨噬細胞%CD36%胞外信號調節激酶通路%炎癥因子
지다당%거서세포%CD36%포외신호조절격매통로%염증인자
Lipopolysaccharide%Macrophage%CD36%Extracellular regulated kinase signaling pathway%Inflammatory cytokine
本研究以鼠源巨噬细胞RAW264．7为模型，研究CD36和胞外信号调节激酶（ERK ）通路对脂多糖（LPS）诱导巨噬细胞分泌炎症因子的影响。首先用100 ng／ml LPS刺激正常及小干扰RNA （siRNA ）技术沉默CD36表达的巨噬细胞16 h ，检测巨噬细胞的ERK活性及分泌炎症因子如肿瘤坏死因子α（TNF‐α）、白细胞介素6（IL‐6）和IL‐10的水平；继而以20 nmol／L ERK抑制剂处理细胞，再用LPS刺激，检测以上各项指标的变化，进一步明确ERK通路与LPS诱导巨噬细胞分泌炎症因子的相关性。结果显示，经LPS刺激，巨噬细胞的ERK活性显著增强，分泌的促炎因子 TNF‐α和 IL‐6显著增高，抑炎因子 IL‐10水平无明显变化；与CD36正常表达的巨噬细胞相比，CD36表达下降的巨噬细胞ERK活性及促炎因子TNF‐α、IL‐6水平显著下降，抑炎因子IL‐10显著增多。与未处理组相比，ERK抑制剂预处理的巨噬细胞中LPS诱导的ERK活性显著降低，促炎因子 TNF‐α和 IL‐6水平降低，抑炎因子 IL‐10水平升高。结果提示，LPS能通过其受体———CD36，激活巨噬细胞内ERK活性，进而促进巨噬细胞促炎因子的分泌。
本研究以鼠源巨噬細胞RAW264．7為模型，研究CD36和胞外信號調節激酶（ERK ）通路對脂多糖（LPS）誘導巨噬細胞分泌炎癥因子的影響。首先用100 ng／ml LPS刺激正常及小榦擾RNA （siRNA ）技術沉默CD36錶達的巨噬細胞16 h ，檢測巨噬細胞的ERK活性及分泌炎癥因子如腫瘤壞死因子α（TNF‐α）、白細胞介素6（IL‐6）和IL‐10的水平；繼而以20 nmol／L ERK抑製劑處理細胞，再用LPS刺激，檢測以上各項指標的變化，進一步明確ERK通路與LPS誘導巨噬細胞分泌炎癥因子的相關性。結果顯示，經LPS刺激，巨噬細胞的ERK活性顯著增彊，分泌的促炎因子 TNF‐α和 IL‐6顯著增高，抑炎因子 IL‐10水平無明顯變化；與CD36正常錶達的巨噬細胞相比，CD36錶達下降的巨噬細胞ERK活性及促炎因子TNF‐α、IL‐6水平顯著下降，抑炎因子IL‐10顯著增多。與未處理組相比，ERK抑製劑預處理的巨噬細胞中LPS誘導的ERK活性顯著降低，促炎因子 TNF‐α和 IL‐6水平降低，抑炎因子 IL‐10水平升高。結果提示，LPS能通過其受體———CD36，激活巨噬細胞內ERK活性，進而促進巨噬細胞促炎因子的分泌。
본연구이서원거서세포RAW264．7위모형，연구CD36화포외신호조절격매（ERK ）통로대지다당（LPS）유도거서세포분비염증인자적영향。수선용100 ng／ml LPS자격정상급소간우RNA （siRNA ）기술침묵CD36표체적거서세포16 h ，검측거서세포적ERK활성급분비염증인자여종류배사인자α（TNF‐α）、백세포개소6（IL‐6）화IL‐10적수평；계이이20 nmol／L ERK억제제처리세포，재용LPS자격，검측이상각항지표적변화，진일보명학ERK통로여LPS유도거서세포분비염증인자적상관성。결과현시，경LPS자격，거서세포적ERK활성현저증강，분비적촉염인자 TNF‐α화 IL‐6현저증고，억염인자 IL‐10수평무명현변화；여CD36정상표체적거서세포상비，CD36표체하강적거서세포ERK활성급촉염인자TNF‐α、IL‐6수평현저하강，억염인자IL‐10현저증다。여미처리조상비，ERK억제제예처리적거서세포중LPS유도적ERK활성현저강저，촉염인자 TNF‐α화 IL‐6수평강저，억염인자 IL‐10수평승고。결과제시，LPS능통과기수체———CD36，격활거서세포내ERK활성，진이촉진거서세포촉염인자적분비。
The present paper aims to establish a RAW264 .7 macrophage model to investigate the effects of CD36 and extracellular regulated kinase (ERK) signaling pathway on the secretion of inflammation‐related cytokines induced by lipopolysaccharide (LPS) .Firstly ,the macrophages with or without CD36 knockdown were stimulated with 100 ng/ml LPS for 16 h .Then the activation of ERK and secretion of tumor necrosis factorα(TNF‐α) ,interleukin 6 (IL‐6) and IL‐10 were analyzed .Secondly ,the cells were incubated with 20 nmol/L ERK inhibitor ,and LPS was added for further stimulation .The changes in the above indexes were investigated .The relationship between the secretion of inflammatory cytokines and ERK signaling pathway was studied .The results showed that in normal RAW264 .7 cells ,LPS treatment stimulated the activation of ERK and secretion of TNF‐α and IL‐6 significantly ,and had no significant effects on IL‐10 level .CD36 knockdown inhibited the activation of ERK and secretion of TNF‐α and IL‐6 , and increased IL‐10 level upon LPS treatment . The pharmaceutical inhibition of ERK decreased TNF‐α and IL‐6 secretion and enhanced IL‐10 secretion upon LPS treatment .The results suggest that both CD36 and ERK pathway are involved in LPS‐mediated secretion of proinflammatory cytokines .
