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目的:观察全反式维甲酸(ATRA)对乙酰胆碱受体(AChR)特异性淋巴细胞的体外调控作用,探讨其治疗重症肌无力(MG)的可能机制。方法:建立完全弗氏佐剂(CFA)对照组及实验性自身免疫性重症肌无力(EAMG)组大鼠,并获取淋巴结单个细胞悬液,以ACh R97-116多肽片段以及不同浓度的ATRA体外培养72 h,采用流式细胞仪法、CCK-8法、ELISA法分别检测活细胞比例、细胞凋亡和周期的改变以及Th亚群的格局和B细胞抗体分泌能力的变化。结果:ATRA显著降低活细胞比例(P<0.001);不同浓度的ATRA均促进了特异性细胞群的凋亡(P<0.001),且呈剂量依赖性,而ATRA未改变AChR特异性淋巴细胞的生长周期;ATRA处理后,CFA和EAMG组的淋巴细胞增殖均受到明显抑制,且ATRA对ACh R特异性的淋巴细胞的抑制明显(EAMG组,P<0.01)于CFA组(P<0.05);ATRA干预后,ACh R特异性CD4+T淋巴细胞的比例下降(P<0.01),且ATRA促进了Th2、Treg细胞亚群百分比(P_(IL-4)<0.001,P_(Foxp3)<0.001),而抑制了促炎性的Th17、Th1细胞亚群百分比(P_(IL-17)<0.05,P_(IFN-γ)<0.001);ATRA能够降低ACh R特异性B细胞的抗体分泌能力(P<0.01)。结论:ATRA不仅能抑制ACh R特异性T细胞功能,同时也能抑制ACh R特异性B细胞功能,其在MG的临床治疗中可能起治疗作用。
AIM: To investigate the in vitro regulation of ATRA on AChR-specific lymphocytes in vitro and to explore its possible mechanism for the treatment of myasthenia gravis (MG). Methods: Complete Freund’s adjuvant (CFA) control group and experimental autoimmune myasthenia gravis (EAMG) group were established. Single lymphocyte suspension was obtained. The ACh R97-116 peptide fragment and different concentrations of ATRA in vitro After cultured for 72 h, the proportion of viable cells, the change of cell cycle and apoptosis, the pattern of Th subpopulation and the secretion of B cell antibody were detected by flow cytometry, CCK-8 and ELISA. Results: ATRA significantly decreased the percentage of viable cells (P <0.001). ATRA at different concentrations promoted the apoptosis of specific cell population (P <0.001) in a dose-dependent manner, while ATRA did not change the percentage of AChR-specific lymphocytes After ATRA treatment, the proliferation of lymphocytes in both CFA and EAMG groups was significantly inhibited. ATRA inhibited the ACh R - specific lymphocytes (P <0.01) in the EAMG group (P <0.05). After ATRA intervention, the percentage of ACh R-specific CD4 + T lymphocytes decreased (P <0.01), and ATRA promoted the percentage of Th2 and Treg subsets (P_ (IL-4) <0.001 and P_ (Foxp3) (P_ (IL-17) <0.05, P_ (IFN-γ) <0.001). ATRA could reduce the antibody-secreting capacity of ACh R-specific B cells (P <0.01). CONCLUSION: ATRA can not only inhibit ACh R-specific T cell function but also inhibit ACh R-specific B cell function, which may play a therapeutic role in the clinical treatment of MG.