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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14d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 Radix Astragali and Astragalus Yupingfeng Powder on the activation of splenic lymphocytes in a mouse model of rapid aging (SAMP8). Methods In vitro experiments were conducted with Astragalus Yupingfeng San, YRF-containing Yupingfeng San, and an equal volume of normal saline intragastric administration of serum-containing mice for 14 days. The serum was incubated with splenic lymphocytes of SAMP8 mice. In vivo experiments, respectively, containing Astragalus Yupingfeng San, YR with YHD, thymosin solution and equal volume of saline administered intragastric SAMP8 mice 14d after blood and spleen to prepare serum and spleen lymphocyte suspension. The level of tumor necrosis factor (TNF-α) was detected by enzyme-linked immunosorbent assay (ELISA) and the expression of CD28 and p38MAPK mRNA in splenic lymphocytes were detected by real-time fluorescence quantitative PCR and Western blotting blot to detect the protein expression. Results Compared with blank serum group and normal saline group, the content of TNF-α in supernatant and serum of drug-containing serum group increased, and the expression of CD28, p38MAPK mRNA and protein in spleen lymphocytes increased. The content of TNF-α and the expression of CD28 and p38MAPK mRNA in HHD-containing Yupingfeng powder group were significantly higher than that in the Yupingfeng group (P <0.01) in vitro. The content of TNF-α (P <0.05), and the expression of CD28 and p38MAPK mRNA in the YHD group was significantly higher than that in the Yupingfeng group (P <0.05). The Chinese herbal medicine group was not as effective as the thymosin group (P <0.05). Conclusion Yupingfeng San with Radix Astragali or Radix Astragali can enhance the activation of splenic lymphocytes in SAMP8 mice and play a role in anti-immune aging.