华中科技大学学报(医学版)
華中科技大學學報(醫學版)
화중과기대학학보(의학판)
ACTA UNIVERSITATIS MEDICINAE TONGJI
2015年
4期
416-418,428
,共4页
谢艳萍%辅桓钦%郭慧慧%姚伟%杨文涛%鲁晟%温晓红
謝豔萍%輔桓欽%郭慧慧%姚偉%楊文濤%魯晟%溫曉紅
사염평%보환흠%곽혜혜%요위%양문도%로성%온효홍
微小RN A%内毒素脂多糖%脓毒症%信号通路
微小RN A%內毒素脂多糖%膿毒癥%信號通路
미소RN A%내독소지다당%농독증%신호통로
micro RNA%lipopolysaccharide%sepsis%signaling pathway
目的:探讨微小RNA155(miR‐155)对脂多糖(LPS)信号通路的调节及可能机制。方法建立miR‐155表达载体模型,体外培养单核巨噬细胞(T HP‐1),分别以超纯水处理T HP‐1细胞(CK组)、LPS处理未经转染的T HP‐1细胞(LPS组)、LPS处理经过miR‐155转染的T HP‐1细胞(LPS+miR‐155组),处理后收集细胞培养液,采用酶联免疫吸附实验(ELISA)检测细胞因子表达水平,Western blot检测TAKl相关结合蛋白2(TAB2)、转化生长因子激酶1(TAK1)蛋白表达情况。结果 LPS+miR‐155组TNF‐α、IL‐6表达水平最低,且显著低于其他两组(均 P<0.05),CK组 TNF‐α、IL‐6表达水平又显著低于 LPS组(均 P<0.05);LPS+ miR‐155组 TAB2表达水平显著低于其他两组(均 P<0.05), LPS组、LPS+miR‐155组TAK1表达水平比较差异无统计学意义(P>0.05)。结论 MiR‐155表达上调具有抑制炎症因子释放的作用,miR‐155可能是通过抑制靶基因T AB2的表达,在调控炎症反应中具有重要作用。
目的:探討微小RNA155(miR‐155)對脂多糖(LPS)信號通路的調節及可能機製。方法建立miR‐155錶達載體模型,體外培養單覈巨噬細胞(T HP‐1),分彆以超純水處理T HP‐1細胞(CK組)、LPS處理未經轉染的T HP‐1細胞(LPS組)、LPS處理經過miR‐155轉染的T HP‐1細胞(LPS+miR‐155組),處理後收集細胞培養液,採用酶聯免疫吸附實驗(ELISA)檢測細胞因子錶達水平,Western blot檢測TAKl相關結閤蛋白2(TAB2)、轉化生長因子激酶1(TAK1)蛋白錶達情況。結果 LPS+miR‐155組TNF‐α、IL‐6錶達水平最低,且顯著低于其他兩組(均 P<0.05),CK組 TNF‐α、IL‐6錶達水平又顯著低于 LPS組(均 P<0.05);LPS+ miR‐155組 TAB2錶達水平顯著低于其他兩組(均 P<0.05), LPS組、LPS+miR‐155組TAK1錶達水平比較差異無統計學意義(P>0.05)。結論 MiR‐155錶達上調具有抑製炎癥因子釋放的作用,miR‐155可能是通過抑製靶基因T AB2的錶達,在調控炎癥反應中具有重要作用。
목적:탐토미소RNA155(miR‐155)대지다당(LPS)신호통로적조절급가능궤제。방법건립miR‐155표체재체모형,체외배양단핵거서세포(T HP‐1),분별이초순수처리T HP‐1세포(CK조)、LPS처리미경전염적T HP‐1세포(LPS조)、LPS처리경과miR‐155전염적T HP‐1세포(LPS+miR‐155조),처리후수집세포배양액,채용매련면역흡부실험(ELISA)검측세포인자표체수평,Western blot검측TAKl상관결합단백2(TAB2)、전화생장인자격매1(TAK1)단백표체정황。결과 LPS+miR‐155조TNF‐α、IL‐6표체수평최저,차현저저우기타량조(균 P<0.05),CK조 TNF‐α、IL‐6표체수평우현저저우 LPS조(균 P<0.05);LPS+ miR‐155조 TAB2표체수평현저저우기타량조(균 P<0.05), LPS조、LPS+miR‐155조TAK1표체수평비교차이무통계학의의(P>0.05)。결론 MiR‐155표체상조구유억제염증인자석방적작용,miR‐155가능시통과억제파기인T AB2적표체,재조공염증반응중구유중요작용。
Objective To explore the regulation of MiR‐155 on LPS signaling pathway and its mechanism.Methods miR‐155 expression vector model was established.Mononuclear cell line THP‐1 was cultured in vitro.The cells were divided into 3 groups:CK group ,THP‐1 cells were treated with ultrapure water ;LPS group ,THP‐1 cells were treated with LPS ;LPS+miR‐155 group ,miR‐155 transfected THP‐1 cells were treated with LPS.After that ,cell culture fluid was collected.Cytokines were determined by enzyme‐linked immunosorbent assay (ELISA) .TAB2 and TAK1 were detected by Western blot.Results Ex‐pression of TNF‐αand IL‐6 was the lowest in LPS+miR‐155 group ,and significantly lower than that in the other two groups(P<0.05).Expression of TNF‐αand IL‐6 was lower in CK group than in LPS group(P<0.05).The level of TAB2 was signifi‐cantly lower in LPS+miR‐155 group than in other two groups(both P<0.05).TAK1 expression level was not different be‐tween LPS group and LPS+miR‐155 group(P>0.05).Conclusion Up‐regulated expression of MiR‐155 can inhibit the release of inflammatory factors.miR‐155 may inhibit TAK1 expression through inhibiting target gene TAB2 expression ,and decrease expression of NF‐κB.miR‐155 plays an important role in regulating inflammatory response.