含红芪、黄芪玉屏风散对SAMP8小鼠脾淋巴细胞活化影响

摘要

目的 比较研究含红芪或黄芪玉屏风散对快速老化模型(SAMP8)小鼠脾淋巴细胞活化的影响.方法 体外实验分别以含黄芪玉屏风散、含红芪玉屏风散及等体积生理盐水灌胃青龄小鼠14 d制备含药血清,将含药血清与SAMP8小鼠脾淋巴细胞共培养.体内实验分别以含黄芪玉屏风散、含红芪玉屏风散、胸腺肽溶液及等体积生理盐水灌胃SAMP8小鼠14 d后取血和脾脏制备成血清及脾淋巴细胞悬液.采用酶联免疫吸附测定(ELISA)检测细胞培养上清液及血清中的肿瘤坏死因子(TNF-α)水平,实时荧光定量PCR检测脾淋巴细胞中CD28分子、p38MAPK mRNA表达及蛋白质印迹法(Western blot)检测其蛋白表达.结果 与空白血清组及生理盐水组比较,含药血清组上清液及血清中TNF-α含量均增高,脾淋巴细胞中CD28分子、p38MAPK mRNA及其蛋白表达量均增高.其中体外实验中含红芪玉屏风散组TNF-α含量及CD28分子、p38MAPK mRNA表达均高于含黄芪玉屏风散组(P<0.01),体内实验中含黄芪玉屏风散组TNF-α含量高于含红芪玉屏风散组(P<0.05),含红芪玉屏风散组CD28分子、p38MAPK mRNA表达均高于含黄芪玉屏风散组(P<0.05),中药组效果均不及胸腺肽组(P<0.05).结论 含红芪或黄芪的玉屏风散对SAMP8小鼠脾淋巴细胞的活化均具有增强作用,起到一定的抗免疫老化作用.%Objective To compare the effects of Yupingfeng San (YPFS, Jade Wind-Barrier Powder) containing Radix Hedysari (sweetvetch root, hongqi) or Radix Astragali (astragalus root, huangqi) on the splenic lymphocyte activation in SAMP8 mice.Methods In vitro experiment: Yupingfeng San containing Radix Hedysari or Radix Astragali and saline of the same volume were given to young mice respectively via intragastric administration for 14 days to prepare dosed serum for co-culture with SAMP8 mice spleen lymphocytes;In vivo experiment: Yupingfeng San containing Radix Hedysari or Radix Astragali, thymus peptide solution and saline of the same volume were given to SAMP8 mice respectively for 14 days before their blood and spleen samples were taken for preparing dosed serum and splenic lymphocyte suspension.Levels of TNF-α in vitro and in vivo were measured with ELISA;expression of CD28 mRNA and p38MAPK mRNA in vitro and in vivo were measured by using real-time PCR and the expression of CD28 and p38MAPK by using Western blot.Results Compared with blank serum group and normal saline group, in vitro and in vivo levels of TNF-α were increased in dosed serum groups.So were the expression of splenic lymphocyte CD28, p38MAPK mRNA and protein expression.In vitro levels of TNF-α and CD28, p38MAPK mRNA expression in the hedysari group were higher than those in the astragali group (P<0.01);in vivo levels of TNF-α and CD28, p38MAPK mRNA expression in the astragali group were higher than those in the hedysari group (P<0.05).CD28 and p38MAPK mRNA expression in the hedysari group were higher than that in the astragali group (P<0.05).Changes of TCM groups were inferior to that of the thymosin group (P<0.05).Conclusion Yupingfeng San containing Radix Hedysari or Radix Astragali both played a potential role in the splenic lymphocyte activation of SAMP8 mice, which serve as a certain resistance to immunosenescence.