纳米雄黄影响MRL/lpr小鼠B细胞表型和功能的初步研究

Realgar nano-particles down-regulated the expression of CD40 in B cells from MRL / lpr mice and promote B cell apoptosis through rising their Ca^(2+) concentra-tion

目的:探索纳米雄黄对MRL/lpr小鼠B细胞功能的影响。方法:分别用生理盐水,低、中、高剂量纳米雄黄混悬液各0.2 ml,给MRL/lpr小鼠灌胃12周后,处死各组小鼠,磁珠分选小鼠脾脏B细胞。CCK-8法检测B细胞增殖活力;流式细胞仪检测B细胞凋亡,Ca2+浓度及CD69、CD40的表达。结果:获得的B细胞纯度高达94%。雄黄不影响MRL/lpr小鼠脾脏B细胞的增殖,也不影响早期活化分子CD69的表达。但雄黄能促进MRL/lpr小鼠脾脏B凋亡,可升高其内Ca2+浓度,并可下调其表面共刺激分子CD40的表达。结论:雄黄能通过升高MRL/lpr小鼠B细胞内Ca2+浓度,促进其凋亡,减少其表面共刺激因子的表达,抑制其活化,从而减少自身抗体的产生,减轻MRL/lpr小鼠自身免疫损伤,减缓SLE病程。

Objective：To explore the effect of Realgar nano-particles on function of B cells obtained from MRL/lpr mouse spleen in vivo. Methods： MRL/lpr mouse were lavaged respectively with physiological saline, low, middle and high dose nano realgar suspension each 0.2 ml, after 12 weeks, the mice was put to death. B cells were sorted from spleen of MRL/lpr mouse using magnetic bead. Proliferation of B cells was tested by CCKS-kit; B cell Ca2＋ concentration, activation and apoptosis of B cells and expression of CD69 and CD40 were both checked by FCM. Results： The purity of sorted B cells was over 94%. Realgar nano-particles influenced neither the proliferation nor the expression of CD69 of B cells from MRL/lpr mice ,while Realgar nano-particles could promote the apop- tosis of B cells from MRL/lpr mice, Furthermore, Realgar nano-partieles could increase its Ca2 ＋concentration, and down -regulated the expression of CD40 in B cells from MRL/lpr mice. Conclusion ： Realgar could promote apoptosis of B cells from MRL/lpr mice through rising Ca2＋ concentration in them, reduce its surface molecule activation and stimulating surface factor expression, restrain the activa- tion, so as to reduce the production of antibodies, reduce MRL/LPR mice autoimmune injury, slow SLE course.