CAJ | 학술논문

背景：类风湿关节炎的病因尚不十分明确，但核转录因子κB在类风湿性关节炎发病中的作用，己逐渐引起风湿病学者的关注。 　　目的：通过RNA干扰技术阻断人类风湿关节炎滑膜细胞核转录因子κB信号通路，探索其在类风湿关节炎治疗中的应用前景。 　　方法：分离、消化、培养滑膜细胞备用。根据小分子干扰RNA设计原则，确定核转录因子κB的小分子干扰RNA 四条目标基因序列，合成并构建核转录因子κB小分子干扰RNA表达载体。将构建好的4条pGenesil-1/核转录因子κB小分子干扰RNA表达载体转染入生长状态良好的一代滑膜细胞，并设立空白和阴性对照组。收集转染后12 h、24 h、48 h、72 h、5 d、7 d不同时间段的细胞，并提取RNA。测定细胞内核转录因子κB mRNA相对表达水平，筛选出有效抑制核转录因子κB mRNA表达的小分子干扰RNA质粒载体。 　　结果与结论：核转录因子κB在人类风湿关节炎滑膜细胞高表达，3#pGenesil-1/核转录因子κB能显著抑制人类风湿关节炎滑膜细胞核转录因子κB mRNA表达。RNA干扰技术可阻断核转录因子κB mRNA的表达，因此，可将阻断核转录因子κB信号通路作为基因治疗类风湿关节炎的靶点。
배경：류풍습관절염적병인상불십분명학，단핵전록인자κB재류풍습성관절염발병중적작용，기축점인기풍습병학자적관주。 　　목적：통과RNA간우기술조단인류풍습관절염활막세포핵전록인자κB신호통로，탐색기재류풍습관절염치료중적응용전경。 　　방법：분리、소화、배양활막세포비용。근거소분자간우RNA설계원칙，학정핵전록인자κB적소분자간우RNA 사조목표기인서렬，합성병구건핵전록인자κB소분자간우RNA표체재체。장구건호적4조pGenesil-1/핵전록인자κB소분자간우RNA표체재체전염입생장상태량호적일대활막세포，병설립공백화음성대조조。수집전염후12 h、24 h、48 h、72 h、5 d、7 d불동시간단적세포，병제취RNA。측정세포내핵전록인자κB mRNA상대표체수평，사선출유효억제핵전록인자κB mRNA표체적소분자간우RNA질립재체。 　　결과여결론：핵전록인자κB재인류풍습관절염활막세포고표체，3#pGenesil-1/핵전록인자κB능현저억제인류풍습관절염활막세포핵전록인자κB mRNA표체。RNA간우기술가조단핵전록인자κB mRNA적표체，인차，가장조단핵전록인자κB신호통로작위기인치료류풍습관절염적파점。
BACKGROUND:The etiological factor for rheumatoid arthritis remains unclear, but the effects of nuclear factor-κB on the onset of rheumatoid arthritis have been gradual y paid great attention by rheumatologists. OBJECTIVE:By using the RNA interference (RNAi) technique to block the signal pathway of nuclear factor-κB mRNA of human rheumatoid arthritis synovial cells, this study explored its application prospect in the treatment of rheumatoid arthritis. METHODS:The synovial cells were isolated, digested, and cultured for further use. In accordance with the design principle of smal interfering RNA (siRNA), target sequences of siRNA of nuclear factor-κB were identified, and siRNA expression vector of nuclear factor-κB was synthesized and constructed. The four pGenesil-1/nuclear factor-κB siRNA expression vectors were transfected into the first passage of synovial cells that wel grew. Blank and negative control groups were set. cells at 12, 24, 48, 72 hours, 5 and 7 days after transfection were col ected, and RNA was extracted. Intracellular nuclear factor-κB mRNA expression levels were measured, and siRNA plasmid vector that could effectively inhibit nuclear factor-κB mRNA expression was screened out. RESULTS AND CONCLUSION:Nuclear factor-κB highly expressed in synovial cells after human rheumatoid arthritis. 3#pGenesil-1/nuclear factor-κB apparently suppressed nuclear factor-κB mRNA expression in synovial cells with human rheumatoid arthritis. RNAi technique blocked nuclear factor-κB mRNA expression. Therefore, the block of nuclear factor-κB signal pathway might be a good target for rheumatoid arthritis gene therapy.