资木瓜总苷抑制RBL-2H3细胞脱颗粒及其机制研究  

Inhibitory effects of Chaenomeles speciosa total saponins on the degranulation of RBL-2H3 mast cells and its mechanism

作　　者：贾云莉 李世刚[1] 柳蔚[1] 喻玲玲[1] 曾小威[1] 

机构地区：[1]三峡大学医学院国家中药药理三级实验室,宜昌443002

出　　处：《免疫学杂志》2015年第10期864-867,共4页Immunological Journal

摘　　要：目的研究资木瓜总苷对RBL-2H3肥大细胞脱颗粒的影响及其作用机制。方法培养大鼠来源的RBL-2H3肥大细胞,资木瓜总苷与RBL-2H3细胞共培养,MTT检测资木瓜总苷对肥大细胞的毒性作用,用荧光分光光度法定量检测RBL-2H3细胞释放的β-己糖胺酶量,计算其释放率,ELISA检测细胞上清TNF-α的释放,流式细胞术检测细胞表面高亲和力Ig E受体FcεRIα的表达,Western blot检测胞内信号Lyn/P-Lyn、PLCγ1/P-PLCγ1的表达。结果与空白对照组相比,资木瓜总苷可显著降低β-己糖胺酶释放率以及TNF-α释放量,降低肥大细胞表面FcεRIα表达,减弱肥大细胞Lyn和PLCγ1磷酸化水平。结论资木瓜总苷可显著抑制RBL-2H3肥大细胞脱颗粒,并存在着明显的量效关系,其作用机制可能是通过Ig E-FcεRIα-Lyn-PLCγ1通路抑制肥大细胞活化脱颗粒。This study aimed to study the effect of total saponins of Chaenomeles speciosa on the degranulation of RBL-2H3 mast cell. RBL-2H3 mast cells from rat was cultured, and then co-cultured with total saponins of Chaenomeles speciosa. MTT method was performed to measure the toxic effects of total saponins of Chaenomeles speciosa on mast cells; fluorescence quantitative spectrophotometric method was used to measure the releasing amount and rate of β- hexosaminidase enzyme; ELISA assay was adopted to test the release of TNF-α in cell supernatants. While flow cytometry was utilized to assay cell surface expression of high affinity Ig E receptor FcεRIα; Western blot was performed to detect the expression of Lyn/P-Lyn, PLCγ1/P-PLCγ1. Compared to the normal control group, the release of β- hexosaminidase and the levels of TNF-α were significantly increase, the expression of cell surface high affinity Ig E receptor FcεRIα was significantly up-regulated, and the phosphorylation levels of intracellular signal protein Lyn and PLCγ1 were significantly elevated in the group stimulated by DNP;whereas, total saponins of Chaenomeles speciosa could significantly decrease the release of β- hexosaminidase and the levels of TNF-α, significantly down-regulate the expression of cell surface high affinity Ig E receptor FcεRIα,and significantly reduce the phosphorylation levels of intracellular signal protein Lyn and PLCγ1. In conclusion,total saponins of Chaenomeles speciosa can inhibit the degranulation of RBL-2H3 mast cells in a dose-dependent manner, and the mechanism may related to the down-regulated expression of FcεRIα, P-Lyn and P-PLCγ1.

关 键 词：资木瓜总苷 RBL-2H3 β-己糖胺酶 TNF-Α FcεRiα 

分 类 号：R285.5[医药卫生—中药学]