中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
5期
844-846
,共3页
树突状细胞%RNA干扰%免疫耐受%基因表达
樹突狀細胞%RNA榦擾%免疫耐受%基因錶達
수돌상세포%RNA간우%면역내수%기인표체
Dendritic cells%RNA interference%Immune tolerance%Gene expression
目的 观察小干扰RNA(siRNA)抑制树突状细胞(DC)中主要组织相容性复合体-Ⅰ(MHC-Ⅰ)类分子的表达,探讨抑制后DC中细胞因子的变化.方法 在最佳干扰浓度,通过流式细胞仪与逆转录-聚合酶链反应(RT-PCR)定量分析方法,选择最能有效抑制DC中MHC-Ⅰ类分子表达的siRNA序列.在对照组、白细胞介素(IL) -10 200 μg,/L组和干扰MHC-Ⅰ表达DC组中,用酶联免疫吸附试验(ELISA)比较IL-12、肿瘤坏死因子(TNF)-α表达量和RT-PCR比较IL-6表达量.结果 在140 nmol/L最佳siRNA转染浓度,siRNA-2干扰组中MHC-Ⅰ的表达量最低.IL-12与TNF-α在空白对照组表达量明显高于IL-10组和siRNA干扰组,差异有统计学意义(P＜0.01)；IL-6在空白对照组的表达量明显低于IL-10组与siRNA干扰组(P＜0.01).结论 在siRNA抑制MHC-Ⅰ表达的DC细胞中,IL-12与TNF-α表达量降低,IL-6表达量升高.
目的 觀察小榦擾RNA(siRNA)抑製樹突狀細胞(DC)中主要組織相容性複閤體-Ⅰ(MHC-Ⅰ)類分子的錶達,探討抑製後DC中細胞因子的變化.方法 在最佳榦擾濃度,通過流式細胞儀與逆轉錄-聚閤酶鏈反應(RT-PCR)定量分析方法,選擇最能有效抑製DC中MHC-Ⅰ類分子錶達的siRNA序列.在對照組、白細胞介素(IL) -10 200 μg,/L組和榦擾MHC-Ⅰ錶達DC組中,用酶聯免疫吸附試驗(ELISA)比較IL-12、腫瘤壞死因子(TNF)-α錶達量和RT-PCR比較IL-6錶達量.結果 在140 nmol/L最佳siRNA轉染濃度,siRNA-2榦擾組中MHC-Ⅰ的錶達量最低.IL-12與TNF-α在空白對照組錶達量明顯高于IL-10組和siRNA榦擾組,差異有統計學意義(P＜0.01)；IL-6在空白對照組的錶達量明顯低于IL-10組與siRNA榦擾組(P＜0.01).結論 在siRNA抑製MHC-Ⅰ錶達的DC細胞中,IL-12與TNF-α錶達量降低,IL-6錶達量升高.
목적 관찰소간우RNA(siRNA)억제수돌상세포(DC)중주요조직상용성복합체-Ⅰ(MHC-Ⅰ)류분자적표체,탐토억제후DC중세포인자적변화.방법 재최가간우농도,통과류식세포의여역전록-취합매련반응(RT-PCR)정량분석방법,선택최능유효억제DC중MHC-Ⅰ류분자표체적siRNA서렬.재대조조、백세포개소(IL) -10 200 μg,/L조화간우MHC-Ⅰ표체DC조중,용매련면역흡부시험(ELISA)비교IL-12、종류배사인자(TNF)-α표체량화RT-PCR비교IL-6표체량.결과 재140 nmol/L최가siRNA전염농도,siRNA-2간우조중MHC-Ⅰ적표체량최저.IL-12여TNF-α재공백대조조표체량명현고우IL-10조화siRNA간우조,차이유통계학의의(P＜0.01)；IL-6재공백대조조적표체량명현저우IL-10조여siRNA간우조(P＜0.01).결론 재siRNA억제MHC-Ⅰ표체적DC세포중,IL-12여TNF-α표체량강저,IL-6표체량승고.
Objective To observe the changes of cytokines in rat dentritic cells (DCs) after suppression of major histocompatibility complex- Ⅰ ( MHC- Ⅰ ) molecules expression by small interfering RNA (siRNA).Methods At the optimal interfering concentration,most effective siRNA sequence suppressing the expression of MHC- Ⅰ molecules in DCs was selected.Enzyme-linked immunosorbent assay (ELISA) was applied to determine interleukin (IL)-12 and tumor necrosis factor (TNF)-α cxpression,and reverse transcription-polymerase chain reaction (RT-PCR) to determine IL-6 in the control,IL-10 (200 μg/L) and.DCs siRNA groups.Results 140 nmol/L siRNA was the optimal interfering concentration.In siRNA-2 group,MHC- Ⅰ was expressed lowest.There was significantly higher expression of IL-12 and TNF-α in the control group than in CDs siRNA group and IL-10 groups ( P ＜0.01 ) and significantly lower level of IL-6 in the control group than in CDs siRNA group and IL-10 group (P ＜0.01 ).Condusion IL-12 and TNF-α expression levels are decreased and IL-6 increased in the DCs after suppression of the MHC-Ⅰ expression with siRNA.
