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目的:探讨RNA干扰(RNAi)后对大鼠T淋巴细胞共刺激分子CD28基因的表达及其对细胞功能的影响。方法:设计针对目标基因的小干扰RNA(siRNA),转染大鼠T淋巴细胞。以FCAS检测CD28水平,MTT检测转染后T淋巴细胞对异体淋巴细胞增殖能力的影响,Realtime-PCR及ELISA法检测细胞因子水平。结果:siRNA转染大鼠T淋巴细胞后可抑制CD28的表达,T细胞增殖能力、IL-2、IFN-γ水平明显低于空白对照组(P<0.05)。结论:siRNA可特异性抑制大鼠T淋巴细胞共刺激分子CD28的表达,降低大鼠T细胞的增殖能力,抑制IL-2、IFN-γ细胞因子的基因水平表达和分泌水平,从而产生免疫耐受效应,为器官移植免疫研究提供实验依据。
Objective: To investigate the effect of RNA interference on the expression of costimulatory molecule CD28 in rat T lymphocytes and its effect on cell function. Methods: Small interfering RNA (siRNA) targeting the target gene was designed and transfected into rat T lymphocytes. The level of CD28 was detected by FCAS. The effect of T lymphocytes on the proliferation of allogeneic lymphocytes was detected by MTT assay. The levels of cytokines were detected by Realtime-PCR and ELISA. Results: siRNA transfected T lymphocytes could inhibit the expression of CD28, the proliferation of T cells, the levels of IL-2 and IFN-γ were significantly lower than those of the blank control group (P <0.05). Conclusion: siRNA can specifically inhibit the expression of costimulatory molecule CD28 in rat T lymphocytes, decrease the proliferation ability of T cells and inhibit the gene expression and secretion of IL-2 and IFN-γ cytokines in rats, resulting in immune tolerance By the effect, provide experimental evidence for organ transplantation immune research.