免疫调节性袋泡茶配方优化及功能性评价

Formulation Optimization and Functional Evaluation of Immunomodulatory Teabags

目的:以绿茶、黄芪、党参、枸杞子、大枣为原料研制一款具有增强免疫力的功能性袋泡茶。方法:以感官评价为指标,应用正交试验结合模糊数学综合评定法优化袋泡茶配方,采用苯酚-硫酸法检测样品中总多糖含量。将SPF级昆明小鼠随机分为空白对照组、模型对照组、阳性对照组(1.40 mg/kg BW的蛋白粉)和袋泡茶低(0.52 g/kg BW)、中(1.04 g/kg BW)、高剂量组(1.56 g/kg BW),除空白对照组外,其余各组小鼠腹腔注射环磷酰胺制备免疫低下模型,连续给药21 d后测定小鼠体重和脏器指数;流式细胞术检测小鼠脾脏淋巴细胞CD3^(+)细胞、CD4^(+)细胞、CD8^(+)细胞及CD4^(+)CD8^(+)细胞比例值;ELISA法测定小鼠血清中干扰素γ(interferon-γ,IFN-γ)含量。结果:袋泡茶的最佳配方为黄芪、党参、枸杞子、大枣、绿茶量分别为茶袋内容物总质量的13.00%、26.30%、20.50%、20.50%、19.70%,多糖含量为33.84±0.52 mg/g。药效实验结果表明,与空白对照组相比,模型对照组小鼠体重和胸腺指数显著降低(P<0.05),CD3^(+)细胞、CD4^(+)细胞、CD8^(+)细胞及CD4^(+)CD8^(+)细胞比例值均显著降低(P<0.05),血清中IFN-γ的含量显著降低(P<0.05),表明免疫抑制小鼠模型建立成功;与模型对照组相比,袋泡茶低剂量组小鼠体重,胸腺指数,CD3^(+)细胞、CD4^(+)细胞、CD8^(+)细胞及CD4^(+)CD8^(+)细胞比例值,血清IFN-γ含量显著升高(P<0.05),袋泡茶中剂量组小鼠胸腺指数,CD4^(+)细胞、CD8^(+)细胞及CD4^(+)CD8^(+)细胞比例值均显著升高(P<0.05);袋泡茶高剂量组小鼠胸腺指数显著升高(P<0.05)。袋泡茶低剂量组效果优于中、高剂量组。结论:袋泡茶具有增强免疫作用,其作用与调节T细胞免疫机制有关。

Objective:The aim of this study was to develop a functional teabags for enhancing immunity with green tea,Astragali radix,Codonopsis radix,Lycii fructus and Jujubae fructus.Methods:Sensory evaluation was used as indicator,the best formula of the teabags was optimized by orthogonal test combined with fuzzy mathematics comprehensive evaluation method.And the total polysaccharide content was detected by the phenol-sulfuric acid method.SPF-grade Kunming mice were randomly divided into blank control group,model control group,positive control group and teabags group with low-,medium-and high-dose(0.52,1.04,1.56 g/kg BW).Except for the blank control group,the immunosuppressive model was prepared by intraperitoneally injected with cyclophosphamidein in all groups,and the body weight and organ index of mice were measured after 21 days of continuous administration.After those processes,CD3^(+)cells,CD4^(+)cells,CD8^(+)cells and CD4^(+)CD8^(+)cells were detected by flow cytometry in the spleen of mice.In addition,the level of interferon-γ(IFN-γ)in mouse serum was detected by ELISA.Results:In the optimal formulation of the teabags,the amounts of Astragali radix,Codonopsis radix,Lycii fructus,Jujubae fructus and green tea were 13.00%,26.30%,20.50%,20.50%and 19.70%of the total mass of the teabags contents,respectively.In addition,the content of polysaccharide was 33.84±0.52 mg/g.The efficacy test results showed that compared with the blank control group,the body weight and thymus index of mice in the model control group were significantly declined(P<0.05),CD3^(+)cells,CD4^(+)cells,CD8^(+)cells and CD4^(+)CD8^(+)cells ratio were significantly reduced(P<0.05),and IFN-γcontent in serum significantly decreased(P<0.05),which indicated that the immunosuppressive model was established successfully.Compared with the model control group,the body weight,thymus index,CD3^(+)cells,CD4^(+)cells,CD8^(+)cells and CD4^(+)CD8^(+)cells ratio,and the serum IFN-γcontent of mice in the low dose group significantly increased(P<0.05).The thymus index,CD4^(+)cells,CD8^(+)cells and CD4^(+)CD8^(+)cells ratio of mice in the medium dose group significantly increased(P<0.05).Additionally,the thymus index of mice in the high dose group significantly increased(P<0.05).Overall,the immunomodulation effect of the teabags in the low-dose group was better than that of the medium-and high-dose group.Conclusion:Immunomodulatory teabags had an immune-enhancing effect,and its effect is related to the regulation of the T cell immune mechanism.