中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
12期
2190-2194
,共5页
MicroRNA-132%肺泡巨噬细胞%炎症反应
MicroRNA-132%肺泡巨噬細胞%炎癥反應
MicroRNA-132%폐포거서세포%염증반응
MicroRNA-132%Alveolar macrophages%Inflammatory response
目的：探讨转染微小RNA-132（miR-132）对肺泡巨噬细胞炎症反应的作用。方法：将体外去致热源培养的大鼠肺泡巨噬细胞株NR8383分为空白对照组、阴性对照组和转染组，分别采用miR-132增敏剂、错义链和PBS作用。转染24 h后，CCK-8法检测细胞増殖；实时荧光定量PCR检测细胞中miR-132的表达；用脂多糖（ LPS）作用细胞后，凝胶电泳迁移率实验（ EMSA ）检测细胞中NF-κB活性；Western blotting法检测细胞中肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）的表达。结果：与空白对照组和阴性对照组相比较，转染组细胞中miR-132的表达明显升高；转染组细胞増殖被明显抑制，与空白对照组相比较，差异有统计学意义（ P＜0.05）；LPS作用后，转染组NF-κB、TNF-α和IL-6表达量显著下降，与空白对照组和阴性对照组相比较，差异均有统计学意义（ P ＜0.05）。结论：转染miR-132可抑制NR8383细胞増殖，并抑制LPS诱导的NR8383细胞的炎症反应。
目的：探討轉染微小RNA-132（miR-132）對肺泡巨噬細胞炎癥反應的作用。方法：將體外去緻熱源培養的大鼠肺泡巨噬細胞株NR8383分為空白對照組、陰性對照組和轉染組，分彆採用miR-132增敏劑、錯義鏈和PBS作用。轉染24 h後，CCK-8法檢測細胞増殖；實時熒光定量PCR檢測細胞中miR-132的錶達；用脂多糖（ LPS）作用細胞後，凝膠電泳遷移率實驗（ EMSA ）檢測細胞中NF-κB活性；Western blotting法檢測細胞中腫瘤壞死因子α（TNF-α）和白細胞介素6（IL-6）的錶達。結果：與空白對照組和陰性對照組相比較，轉染組細胞中miR-132的錶達明顯升高；轉染組細胞増殖被明顯抑製，與空白對照組相比較，差異有統計學意義（ P＜0.05）；LPS作用後，轉染組NF-κB、TNF-α和IL-6錶達量顯著下降，與空白對照組和陰性對照組相比較，差異均有統計學意義（ P ＜0.05）。結論：轉染miR-132可抑製NR8383細胞増殖，併抑製LPS誘導的NR8383細胞的炎癥反應。
목적：탐토전염미소RNA-132（miR-132）대폐포거서세포염증반응적작용。방법：장체외거치열원배양적대서폐포거서세포주NR8383분위공백대조조、음성대조조화전염조，분별채용miR-132증민제、착의련화PBS작용。전염24 h후，CCK-8법검측세포증식；실시형광정량PCR검측세포중miR-132적표체；용지다당（ LPS）작용세포후，응효전영천이솔실험（ EMSA ）검측세포중NF-κB활성；Western blotting법검측세포중종류배사인자α（TNF-α）화백세포개소6（IL-6）적표체。결과：여공백대조조화음성대조조상비교，전염조세포중miR-132적표체명현승고；전염조세포증식피명현억제，여공백대조조상비교，차이유통계학의의（ P＜0.05）；LPS작용후，전염조NF-κB、TNF-α화IL-6표체량현저하강，여공백대조조화음성대조조상비교，차이균유통계학의의（ P ＜0.05）。결론：전염miR-132가억제NR8383세포증식，병억제LPS유도적NR8383세포적염증반응。
[ ABSTRACT] AIM:To investigate the effect of microRNA-132 ( miR-132 ) transfection on the lipopolysaccharide ( LPS)-induced inflammation in rat alveolar macrophages.METHODS:The rat alveolar macrophage NR8383 cultured with-out pyrogen in vitro were divided into blank control group, negative control group and transfected group.The cells in the 3 groups were transfected with phosphate buffer solution ( PBS) , Lipofectamine 2000 and synthesized miR-132 mimic respec-tively.The cell proliferation was detected by Cell Counting Kit-8 ( CCK-8) assay.Real-time PCR was used to detect the ex-pression of miR-132 in the cells.After NR8383 cells were stimulated with LPS for 6 h, the NF-κB DNA-binding activity was measured by electrophoretic mobility shift assay ( EMSA) .The expression of tumor necrosis factor-α( TNF-α) and interleu-kin-6 (IL-6) in NR8383 cells was assayed by Western blotting.RESULTS: After transfection, the expression of miR-132 was significantly higher than that in blank control group and negative control group.The growth of NR8383 cells in transfect-ed group was significantly inhibited compared with blank control group and negative control group ( P<0.05 ) .After the cells were stimulated with LPS, the productions of NF-κB, TNF-αand IL-6 in transfected NR8383 cells were decreased com-pared with blank control group and negative control group (P<0.05).CONCLUSION:Transfection of alveolar macropha-ges with miR-132 significantly suppresses the cell growth, and inhibits inflammatory responses induced by LPS.
