【摘要】：目的:研究干擾素誘導蛋白16(IFI16)表達抑制后對人主動脈外膜成纖維細胞(HAAFs)凋亡的影響。方法:應用IFI16基因小干擾RNA(siRNA)轉染HAAFs使IFI16基因沉默(IFI16-siRNA組),以非特異性siRNA轉染作為其轉染陰性對照組(Con-siRNA組),未經(jīng)處理的HAAFs作為未干預陰性對照組(Neg組)。應用流式細胞儀檢測細胞周期及凋亡,實時定量逆轉錄-聚合酶鏈反應(Realtime qRT-PCR)檢測細胞中IFI16 mRNA表達變化,蛋白免疫印跡(Western blotting)檢測IFI16、抑癌因子(p53)、細胞周期依賴性蛋白激酶抑制因子p21~(WAF)(p21)、B細胞淋巴瘤-2(Bcl-2)相關X蛋白(Bax)及Bcl-2蛋白表達。結果:IFI16-siRNA組與Con-siRNA組、Neg組相比,細胞凋亡率[(3.33±0.41)%vs(7.42±1.51)%(、6.49±1.10)%,P0.05]與G_0/G_1期細胞比例[(56.64±4.77)%vs(69.67±3.54)%、(68.29±4.14)%,P0.05]減少;而S期細胞比例[(25.23±5.19)%vs(13.76±2.07)%、(14.04±3.00)%,P0.05]增加;伴隨著IFI16mRNA表達減少(P0.05),以及IFI16-siRNA組IFI16、p53、p21、Bax蛋白表達量減少(P0.05);同時Bcl-2蛋白表達量增加(P0.05)。結論:抑制IFI16表達可減少HAAFs細胞凋亡,促進細胞周期G_1/S轉換,其機制部分與抑制p53/p21信號通路及調(diào)控Bax/Bcl-2表達有關。
[Abstract]:Aim: to investigate the effect of inhibition of interferon-induced protein 16 (IFI16) expression on (HAAFs) apoptosis in human aortic adventitial fibroblasts. Methods: IFI16 gene was silenced by IFI16 gene small interfering RNA (siRNA) transfection into HAAFs (IFI16-siRNA group), non-specific siRNA transfection was used as the negative control group (Con-siRNA group), and untreated HAAFs was used as unintervention negative control group (Neg group). Flow cytometry was used to detect cell cycle and apoptosis, real-time quantitative reverse transcription-polymerase chain reaction (Realtime qRT-PCR) was used to detect the expression of IFI16 mRNA, and Western blot (Western blotting) was used to detect IFI16, tumor suppressor factor (p53). Cell cycle dependent protein kinase inhibitor p21 ~ (WAF) (p21), B cell lymphoma-2 (Bcl-2)-related X protein (Bax) and Bcl-2 protein were expressed. Results: compared with Con-siRNA group and Neg group, apoptosis rate of IFI16-siRNA group was (3.33 鹵0.41)% vs (7.42 鹵1.51)% (6.49 鹵1.10)%. The proportion of cells in G_0/G_1 phase [(56.64 鹵4.77)% vs, (69.67 鹵3.54)%, (68.29 鹵4.14)%, P0.05] was decreased. The percentage of S phase cells increased [(25.23 鹵5.19)% vs (13.76 鹵2.07)%, (14.04 鹵3.00)%, P0.05]. With the decrease of IFI16mRNA expression (P0.05) and the decrease of IFI16,p53,p21,Bax protein expression in IFI16-siRNA group (P0.05), the expression of Bcl-2 protein increased (P0.05). Conclusion: inhibiting the expression of IFI16 can reduce the apoptosis of HAAFs cells and promote the cell cycle 1 / S transition. The mechanism is partly related to the inhibition of p53/p21 signaling pathway and the regulation of Bax/Bcl-2 expression.
【作者單位】： 貴州省人民醫(yī)院貴州醫(yī)科大學附屬人民醫(yī)院心內(nèi)科;
【基金】：國家自然科學基金資助項目(81260030) 貴州省科技合作計劃項目(黔科合LH字[2015]7159號)
【分類號】：R54

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