CAJ | 학술논문

目的：设计并筛选能高效抑制创伤性关节炎滑膜细胞肿瘤坏死因子α（T N F‐α）表达的最佳干扰序列，为进一步研究TNF‐α基因对创伤性关节炎的临床应用奠定基础。方法根据人源 TNF‐α mRNA 的序列，设计合成3组不同的且能干扰 TNF‐α基因表达的小型干扰 RNA（siRNA）序列，将其转染人创伤性关节炎滑膜细胞，与阴性对照组（NC 组）比较。培养48 h 后，采用实时灾光定量 PCR（RT‐PCR）检测滑膜细胞 TNF‐α mRNA 表达水平，采用 ELISA 法检测细胞上清液 TNF‐α表达水平。结果与 NC 组相比，3组 siRNA 中有2组 siRNA 可有效使人滑膜细胞中 TNF‐α mRNA 表达减少（P＜0．01）；同时细胞上清液中 TNF‐α分泌量下降，与 NC 组比较差异有统计学意义（P＜0．01）。结论成功设计合成了特异且高效阻断 TNF‐α表达的 siRNA 。
목적：설계병사선능고효억제창상성관절염활막세포종류배사인자α（T N F‐α）표체적최가간우서렬，위진일보연구TNF‐α기인대창상성관절염적림상응용전정기출。방법근거인원 TNF‐α mRNA 적서렬，설계합성3조불동적차능간우 TNF‐α기인표체적소형간우 RNA（siRNA）서렬，장기전염인창상성관절염활막세포，여음성대조조（NC 조）비교。배양48 h 후，채용실시재광정량 PCR（RT‐PCR）검측활막세포 TNF‐α mRNA 표체수평，채용 ELISA 법검측세포상청액 TNF‐α표체수평。결과여 NC 조상비，3조 siRNA 중유2조 siRNA 가유효사인활막세포중 TNF‐α mRNA 표체감소（P＜0．01）；동시세포상청액중 TNF‐α분비량하강，여 NC 조비교차이유통계학의의（P＜0．01）。결론성공설계합성료특이차고효조단 TNF‐α표체적 siRNA 。
Objective To design and identify small interfering RNA (siRNA) targeting tumor necrosis factor‐α (TNF‐α) ex‐pression ,siRNAs were electroporated into synovial cells of Human to screen those which can effectively suppress TNF‐α expres‐sion .Methods Three TNF‐α specific double stranded siRNA were designed targeting different regions of TNF‐α mRNA(compared with negative control group) .RT‐PCR and Elisa were applied to detect TNF‐α ,mRNA expression and the secretion of TNF‐α in cell supernatant ,respectively .Results Two of the three customized TNF‐α siRNA could inhibit the expression of TNF‐α mRNA in synovial cells of Human (P< 0 .01) .At the same time ,the secretion of TNF‐α decreased in cell supernatant ,the difference was sig‐nificant statistically compared with the control group(P< 0 .01) .Conclusion TNF‐α siRNA can be successfully designed and syn‐thesized ,which can specifically and effectively suppress TNF‐α mRNA expression .