参杞酵素发酵工艺优化及调节免疫功能评价

Optimization of Fermentation Technology and Evaluation of Regulating Immune Function of Ginseng and Wolfberry Fermentation Liquid

目的:优化参杞酵素的发酵工艺,并研究参杞酵素对小鼠免疫功能的调节作用。方法:以总酸含量为响应值,以发酵时间、发酵温度、菌种比例、接种量为自变量,通过单因素实验和Box-Behnken试验优化发酵工艺。以80 mg/(kg·d)环磷酰胺诱导小鼠构建免疫低下小鼠模型,分别用低(5.0 mL/kg)、中(10.0 mL/kg)、高(15.0 mL/kg)剂量的试验酵素液免疫调节干预,模型组和正常组灌胃等体积的蒸馏水,测定血清中免疫球蛋白G(Immunoglobulin G,IgG)含量和白细胞介素-4(Interleukin-4,IL-4)、白细胞介素-10(Interleukin-10,IL-10)、干扰素-γ(Interferon-γ,IFN-γ)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)的相对表达量,利用碳廓清法检测小鼠吞噬细胞的吞噬能力及对脏器指数的影响。结果:优化得到发酵工艺为发酵时间72 h,发酵温度37℃,接种量5%,菌种比例5:4.5:0.5,总酸含量为1.23 g/100 g。试验酵素可显著(P<0.05)上调免疫抑制小鼠的血清中IFA-γ、TNF-α、IL-4及Ig G水平,下调IL-10水平,高剂量的小鼠血清IL-10质量浓度可恢复至72.26 pg/mL,小鼠血清IFN-γ质量浓度达到213.47 pg/mL,TNF-α质量浓度可达246.13 pg/mL,IgG的质量浓度可达119.89 ng/mL,接近或高于正常组,明显增强吞噬细胞的吞噬能力。结论:参杞酵素对小鼠免疫功能具有正向调节作用。

Objective:To optimize the fermentation process of ginseng and wolfberry fermentation liquid and study its regulating effect on the immune function of mice.Methods:The fermentation process was optimized by single factor test and Box-Behnken test with total acid content as response value and fermentation time,fermentation temperature,strain proportion and inoculum amount as independent variables.The mice were induced with 80 mg/(kg·d)cyclophosphamide to establish an immunocompromised mouse model.The mice were treated with low(5.0 mL/kg),medium(10.0 mL/kg)and high(15.0 mL/kg)doses of experimental fermentation liquid solution,respectively.The contents of immunoglobulin G(IgG),interleukin-4(IL-4),interleukin-10(IL-10)and interferon-γ(interferon-γ)in serum were determined.The relative expression levels of IFN-γand tumor necrosis factor-α(TNF-α)were measured.The phagocytosis ability of mouse phagocytes and its effect on organ index were detected by carbon clearance method.Results:The fermentation process was optimized as follows:Fermentation time 72 h,fermentation temperature 37℃,inoculation amount 5%,strain ratio 5:4.5:0.5,total acid content 1.23 g/100 g.The levels of IFA-γ,TNF-α,IL-4 and IgG in the serum of immunosuppressed mice were significantly up-regulated by the fermentation liquid,and the levels of IL-10 were down-regulated.The mass concentration of IL-10 and IFN-γin the serum of mice with high dose could be restored to 72.26 and 213.47 pg/mL,respectively.The mass concentration of TNF-αand IgG reached 246.13 pg/mL and 119.89 ng/mL,which were close to or higher than those of solvent control group, and significantly enhanced the phagocytic ability of phagocytic cells. Conclusion: The ginseng and wolfberry fermented liquid had a positive regulatory effect on the immune function of mice.