一种中药单体小复方对变应性鼻炎模型小鼠的免疫调节作用  

Immunoregulatory mechanisms of an optimal Chinese herbal monomer compound in mice with allergic rhinitis

作　　者：程坚[1] 张新民[1] 沈自尹[1] 陈伟华[1] 蔡外娇[1,2] 应健[1] 胡国让 刘闰红[1] 

机构地区：[1]复旦大学华山医院中西医结合研究所,上海200040 [2]The Buck Institute [3]Department of Immunology and Allergy Concord Hospital the University of Sydney

出　　处：《中西医结合学报》2011年第12期1360-1366,共7页Journal of Chinese Integrative Medicine

基　　金：上海市科学技术委员会攻关项目(No.02DZ19128)

摘　　要：目的:探讨3种中药单体淫羊藿苷、黄芩苷、黄芪甲苷经优化配比组成的小复方对变应性鼻炎模型小鼠的免疫调节作用。方法:采用卵清蛋白腹腔注射与滴鼻结合的方法建立变应性鼻炎小鼠模型,分离小鼠脾淋巴细胞后体外培养,并将细胞分为空白对照组、刀豆素A组、复方组3组,在不同时间点运用细胞增殖-毒性检测试剂盒连续观察各组细胞的增殖情况,流式细胞术检测各组细胞周期变化,荧光定量分析仪结合荧光探针Fluo-3检测细胞内游离钙离子浓度变化。结果:中药单体小复方可以有效抑制刀豆素A诱导的变应性鼻炎小鼠脾淋巴细胞增殖(P<0.01),且呈现出一定的时效关系,抑制率随时间的延长而增强,在48h达最大值。流式细胞术检测各组细胞的细胞周期变化发现,刀豆素A可以通过降低G0/G1期百分比,升高S期与G2/M期的百分比而促进细胞进入分裂期,复方组与刀豆素A组相比,则在4个时间点均可以升高G0/G1期比例,并降低S期与G2/M期的比例,作用在24h最显著(P<0.05或P<0.01)。荧光定量分析仪检测发现,前24h,刀豆素A诱导后的淋巴细胞荧光强度随时间延长而逐渐增强,而复方组可以明显抑制刀豆素A导致的荧光强度升高,降低细胞内游离钙离子浓度(P<0.01)。结论:中药单体小复方可以抑制刀豆素A诱导的变应性鼻炎模型小鼠脾淋巴细胞增殖,其作用机制可能为通过降低细胞内游离钙离子浓度,从而抑制细胞由G0/G1期进入S和G2/M期。Objective： To study the immunoregulatory effect of an optimal Chinese herbal monomer compound, which consists of three monomers, namely, icariin, baicalin and Astragalus saponin I, in a mouse model of allergic rhinitis.Methods： A mouse model of allergic rhinitis was established by intraperitoneal injection of ovalbumin and aluminum hydroxide gel suspension. The splenic lymphocytes of the mice were separated, cultured in 96-well plates and divided into three groups： control group, concanavalin A group and compound group. Splenic lymphocyte proliferation was detected by cell counting kit-8 method at different time points. Cell cycle distribution was observed by flow cytometry （FCM） also at different time points. The changes of intracellular calcium concentration of splenic lymphocytes were measured by fluorescence microplate reader after the cells were incubated with fluorescence probe Fluo-3/AM. Results： The Chinese herbal monomer compound could inhibit cell proliferation induced by concanavalin A （P〈0.01）. And the inhibition presented a time-effect relationship. With extending of the action time, the inhibition rate gradually increased and reached peak at the 48th hour. FCM test revealed the fact that concanavalin A could promote cells to enter into the mitosis by reducing the percentage of cells in G0/G1 phases while increasing the percentage of cells in S and G2/M phases. Compared with the concanavalin A, the compound could increase the percentage of cells in G0/G1 phases and at the same time reduce the percentage of cells in S and G2/M phases at different time points, with the effect most significant at the 24th hour （P〈0.05 or P〈0. 01）. The results of the test taken by the fluorescence microplate reader revealed that the fluorescence value of the concanavalin A group increased with time in the previous 24 h while the compound could reduce this trend obviously, thus reduce the intracellular calcium concentration （P〈0.01）. Conclusion： The Chinese herbal monomer compound can inh

关 键 词：中草药 复方 鼻炎 变应性 淋巴细胞 细胞周期 细胞内游离钙离子 小鼠 

分 类 号：R285.5[医药卫生—中药学]