目的 探討參麥注射液(SMI)對(duì)RBL-2H3細(xì)胞脫顆粒的影響及其原因。方法 以C48/80為工具藥建立RBL-2H3細(xì)胞脫顆粒模型, 檢測(cè)不同濃度C48/80與RBL-2H3細(xì)胞作用不同時(shí)間后細(xì)胞β-己糖苷酶、類胰蛋白酶和組胺的釋放率以及細(xì)胞活力, 在細(xì)胞活力大于80%情況下, 選擇釋放程度較高的指標(biāo)和條件為優(yōu)選考察指標(biāo)和條件。將SMI和其溶劑(Tween-80)原液等比稀釋成不同濃度后與RBL-2H3細(xì)胞共同培養(yǎng), 通過中性紅染色法觀察細(xì)胞脫顆粒的形態(tài)學(xué)變化, 分別用顯色法和間接熒光法檢測(cè)細(xì)胞上清的β-己糖苷酶和組胺釋放率, 采用細(xì)胞計(jì)數(shù)法(CCK-8)檢測(cè)細(xì)胞的活力。結(jié)果 RBL-2H3細(xì)胞脫顆粒模型的最佳作用時(shí)間為30 min, 最佳指標(biāo)為β-己糖苷酶和組胺釋放率。與空白組相比, SMI質(zhì)量濃度低于13.3 g生藥/L(3倍臨床濃度)時(shí), 細(xì)胞中性紅染色未見脫顆?，F(xiàn)象, 組胺和β-己糖苷酶釋放率亦無差異;而在Tween-80質(zhì)量濃度為1.00 g/L時(shí), SMI 40 g生藥/L(9倍臨床濃度)組和溶劑1.00 g/L組細(xì)胞中性紅染色均可見脫顆?，F(xiàn)象, 細(xì)胞上清組胺和β-己糖苷酶釋放率亦明顯增加。此外, CCK-8結(jié)果顯示, 與空白組相比, 各濃度的SMI對(duì)細(xì)胞活力均無影響。結(jié)論 SMI低于3倍臨床濃度無明顯RBL-2H3細(xì)胞脫顆粒作用;而在9倍臨床濃度能刺激細(xì)胞脫顆粒, 這種脫顆粒作用可能與所含溶劑(Tween-80)有關(guān), 與其對(duì)RBL-2H3的細(xì)胞毒性作用無關(guān)。提示SMI在低于3倍臨床濃度相對(duì)安全, 在9倍臨床濃度時(shí)有致類過敏反應(yīng)的風(fēng)險(xiǎn)。

Objective To explore the effect on degranulation of RBL-2H3 cells and its reasons induced by Shenmai Injection (SMI). Methods Compound 48/80 (C48/80) was used to establish RBL-2H3 cells degranulation model, the release rates of β-hexosaminidase, Tryptase, and histamine and the cell viability of RBL-2H3 cells were detected after treated by different times at the different concentration of C48/80; The indicators and conditions which had a higher degree of release were selected as the preferred study indicators and conditions in the case of that the cell viability was more than 80 percents. RBL-2H3 cells were cultured with the geometric dilution at different concentrations of the stock solution of SMI and its solvent (Tween-80), the morphology of degranulation was observed by neutral red staining. In addition, the releasing rates of β-hexosaminidase and histamine in cell supernatants were detected by chromogenic assay and indirect fluorescence analysis, respectively; The viability of RBL-2H3 cells was tested by cell counting kits. Results The optimal condition of C48/80-induced RBL-2H3 cells degranulation test was 30 min and the best indicators were the release rates of β-hexosaminidase and histamine. When compared with the control group, the release rates of histamine and β-hexosaminidase were no difference and the degranulation by neutral red staining did not appear in SMI groups with the concentrations less than 13.3 g crude drug/L (3 times as much as the clinical concentration) while the release rates of histamine and β-hexosaminidase were significantly increased and the degranulation by neutral red staining appeared in SMI 40 g crude drug/L (9 times as the clinical concentration) group and SMI solvent group that both of them included 1.00 g/L Tween-80. In addition, the results of CCK-8 showed that there was no significant effect in different SMI doses groups on cell viability compared with the control group. Conclusion SMI at the concentration below 3 times as much as the clinical concentration has no significant degranulation of RBL-2H3 cells while at the concentration of 9 times as much as the clinical concentration shows the certain effect on inducing the degranulation of cells which may be related to its solvent that contains Tween-80 and shows no relationship with its toxic effects on the cells. It suggests that SMI is relatively safe at the concentration below 3 times as much as the clinical concentration while it has the risk of allergic reactions at the concentration of 9 times as much as the clinical concentration.

重大新藥創(chuàng)制科技重大專項(xiàng)(2013ZX09302303);重大新藥創(chuàng)制科技重大專項(xiàng)(2012ZX09301003-001-008);國(guó)家自然基金(81001254);河北省科技計(jì)劃項(xiàng)目(13272503D);北京市科委項(xiàng)目基金(Z131100006513010)