玉屏风散和复芪止汗颗粒对免疫抑制小鼠免疫功能的影响
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摘要
目的：用环磷酰胺制备免疫低下小鼠模型,通过测定各组小鼠胸腺指数、脾脏指数、腹腔巨噬细胞吞噬能力、NK细胞杀伤活性、血清溶血素水平、T、B淋巴细胞亚群、血清IL-1β、IL-4、IFN-γ水平,探讨玉屏风散和复芪止汗颗粒高中低各个剂量组对各组小鼠固有免疫和适应性免疫的影响,进而比较玉屏风散和复芪止汗颗粒在提高小鼠免疫力方面的差异,以期为临床应用提供一定的实验基础。
方法：采用标准昆明种小鼠120只,雌雄各半,应用随机数字表法分为8组：生理盐水组、环磷酰胺组、玉屏风散高剂量组(玉高组),玉屏风散中剂量组(玉中组),玉屏风散低剂量组(玉低组),复芪止汗高剂量组(复高组),复芪止汗中剂量组(复中组),复芪止汗低剂量组(复低组),除正常组外,其余各组小鼠均连续7天腹腔注射环磷酰胺制备免疫力低下小鼠模型,造模同时,生理盐水组、环磷酰胺组每天灌胃生理盐水、玉屏风散高剂量(12g/kg·d)、玉屏风散中剂量(6g/kg·d)、玉屏风散低剂量(3g/kg·d)剂量和复芪止汗颗粒高剂量(54g/kg·d)、复芪止汗颗粒中剂量(27g/kg·d)、复芪止汗颗粒低剂量(13.5g/kg·d)剂量各自灌胃,连续14天,精确测定胸腺、脾脏质量计算出小鼠胸腺指数、脾指数；腹腔巨噬细胞吞噬中性红法检测单核-巨噬细胞的吞噬功能；MTT法检测T淋巴细胞增殖功能；乳酸脱氢酶释放法测定NK细胞杀伤活性；流式细胞仪检测CD3+、 CD4+、CD8+T淋巴细胞及CD19+B淋巴细胞水平；半数溶血值测定法检测血清溶血素含量；ELISA法测定血清IL-1β、IL-4和IFN-y的含量。
结果：1,玉屏风散和复芪止汗颗粒各个剂量组均能不同程度的提高被环磷酰胺抑制了的小鼠固有免疫能,与生理盐水组相比,低剂量组在提高巨噬细胞吞噬指数、NK细胞杀伤活性、血清IL-1β水平方面具有统计学意义,高剂量组在提高小鼠NK细胞杀伤活性方面亦具有统计学意义(P<0.05),与环磷酰胺组相比,除低剂量组不能提高小鼠NK细胞杀伤活性和血清IL-1β水平外(P>0.05),其余各组和环磷酰胺组相比均具有统计学意义(P<0.05)。2,两复方对适应性免疫功能的影响方面,环磷酰胺组CD191、IL-4、血清溶血素水平、IFN-γ、T淋巴细胞增殖指数、T淋巴细胞亚群方面与生理盐水组相比具有统计学意义(P<0.05),低剂量组在提高CD19+、IL-4、血清溶血素水平、IFN-y方面与生理盐水组相比亦具有统计学意义,此外,除低剂量组与环磷酰胺组相比不能提高免疫抑制小鼠CD19+、IL-4.血清溶血素水平外(P>0.05),高、中剂量组均可不同程度的提高小鼠免疫功能(P<0.05)。
结论：玉屏风散和复芪止汗颗粒均能不同程度的提高免疫抑制小鼠的免疫功能,表明其作用相似,但两复方在提高细胞免疫方面疗效较强,且以中剂量组效果最好。
Objective:To discuss the influence of Yupingfengsan and Fuqizhihan granule on inherent immunity and adaptive immunity in mice, compare the difference of Yupingfengsan and Fuqizhihan granule in improving mice immunity, and provide some experimental basis for clinical application.
Methods:Kunming mice (60male and60female) were randomly divided into8groups:normal group, model group, Yupingfengsan high dose group, Yupingfengsan middle dose group, Yupingfengsan low dose group,Fuqizhihankeli high dose group,Fuqizhihankeli middle dose group,Fuqizhihankeli low dose group.Make immunosuppression model mice through introperitoneal inject cyclophosphamide except normal group for seven days. gavage normal saline to normal group and model group, Yupingfengsan to Yupingfengsan high, middle, low dose group and Fuqizhihan granule to Fuqizhihan granule high, middle, low dose group once a day respectively for14days. determine the weight of thymus and spleen precisely and calculate the immune organ index; detect the phagocytosis of monocyte-macrophages through neutral red method, proliferation of T lymphocyte by MTT assay, killing activity of natural killer cells by LDH-release method, contents of CD3+, CD4+, CD8+, CD19+on flow cytometry, contents of serum hemolysin by HC50assay, serum contents of IL-1, IL-4and IFN-y by ELISA.
Results:1, Every dose group in Yupingfengsan and Fuqizhihan granule can improve innate immunity of Immunosuppressed mice in different degree. Compared with saline group, all indexes in cyclophosphamide group were statistically significant difference, besides, low dose group could improve killing activity of natural killer cells, phagocytosis index, serum IL-1β,in certain extent, but compared with saline group had statistically significant difference (P<0.05). Compared with cyclophosphamide group, all index except killing activity of natural killer cells and IL-1β had statistically difference.2, The influence of two compounds on adaptive immunity. CD19+,IL-4serum hemolysin, IFN-y, proliferation of T lymphocyte and contents of CD3+, CD4+, CD8+in cyclophosphamide group had statistically significant difference compared with saline group (P<0.05). low dose group could improve CD19+,IL-4serum hemolysin, IFN-y in some extent, but compared with saline group had statistically significant difference(P<0.05). Besides, compared with cyclophosphamide group low dose group could not improve CD19+,IL-4serum hemolysin, IFN-y (P>0.05), but high, middle dose group could improve adaptive immunity in some extent (P<0.05)
Conlusion:Both Yupingfengsan and Fuqizhihan granule could improve immunosuppressive mice immunity, the result revealed that their effect was similar. Both two compounds were superior in restoring cell immunity for immunosuppressive mice. Middle dose group had the best effect.
方法：采用标准昆明种小鼠120只,雌雄各半,应用随机数字表法分为8组：生理盐水组、环磷酰胺组、玉屏风散高剂量组(玉高组),玉屏风散中剂量组(玉中组),玉屏风散低剂量组(玉低组),复芪止汗高剂量组(复高组),复芪止汗中剂量组(复中组),复芪止汗低剂量组(复低组),除正常组外,其余各组小鼠均连续7天腹腔注射环磷酰胺制备免疫力低下小鼠模型,造模同时,生理盐水组、环磷酰胺组每天灌胃生理盐水、玉屏风散高剂量(12g/kg·d)、玉屏风散中剂量(6g/kg·d)、玉屏风散低剂量(3g/kg·d)剂量和复芪止汗颗粒高剂量(54g/kg·d)、复芪止汗颗粒中剂量(27g/kg·d)、复芪止汗颗粒低剂量(13.5g/kg·d)剂量各自灌胃,连续14天,精确测定胸腺、脾脏质量计算出小鼠胸腺指数、脾指数；腹腔巨噬细胞吞噬中性红法检测单核-巨噬细胞的吞噬功能；MTT法检测T淋巴细胞增殖功能；乳酸脱氢酶释放法测定NK细胞杀伤活性；流式细胞仪检测CD3+、 CD4+、CD8+T淋巴细胞及CD19+B淋巴细胞水平；半数溶血值测定法检测血清溶血素含量；ELISA法测定血清IL-1β、IL-4和IFN-y的含量。
结果：1,玉屏风散和复芪止汗颗粒各个剂量组均能不同程度的提高被环磷酰胺抑制了的小鼠固有免疫能,与生理盐水组相比,低剂量组在提高巨噬细胞吞噬指数、NK细胞杀伤活性、血清IL-1β水平方面具有统计学意义,高剂量组在提高小鼠NK细胞杀伤活性方面亦具有统计学意义(P<0.05),与环磷酰胺组相比,除低剂量组不能提高小鼠NK细胞杀伤活性和血清IL-1β水平外(P>0.05),其余各组和环磷酰胺组相比均具有统计学意义(P<0.05)。2,两复方对适应性免疫功能的影响方面,环磷酰胺组CD191、IL-4、血清溶血素水平、IFN-γ、T淋巴细胞增殖指数、T淋巴细胞亚群方面与生理盐水组相比具有统计学意义(P<0.05),低剂量组在提高CD19+、IL-4、血清溶血素水平、IFN-y方面与生理盐水组相比亦具有统计学意义,此外,除低剂量组与环磷酰胺组相比不能提高免疫抑制小鼠CD19+、IL-4.血清溶血素水平外(P>0.05),高、中剂量组均可不同程度的提高小鼠免疫功能(P<0.05)。
结论：玉屏风散和复芪止汗颗粒均能不同程度的提高免疫抑制小鼠的免疫功能,表明其作用相似,但两复方在提高细胞免疫方面疗效较强,且以中剂量组效果最好。
Objective:To discuss the influence of Yupingfengsan and Fuqizhihan granule on inherent immunity and adaptive immunity in mice, compare the difference of Yupingfengsan and Fuqizhihan granule in improving mice immunity, and provide some experimental basis for clinical application.
Methods:Kunming mice (60male and60female) were randomly divided into8groups:normal group, model group, Yupingfengsan high dose group, Yupingfengsan middle dose group, Yupingfengsan low dose group,Fuqizhihankeli high dose group,Fuqizhihankeli middle dose group,Fuqizhihankeli low dose group.Make immunosuppression model mice through introperitoneal inject cyclophosphamide except normal group for seven days. gavage normal saline to normal group and model group, Yupingfengsan to Yupingfengsan high, middle, low dose group and Fuqizhihan granule to Fuqizhihan granule high, middle, low dose group once a day respectively for14days. determine the weight of thymus and spleen precisely and calculate the immune organ index; detect the phagocytosis of monocyte-macrophages through neutral red method, proliferation of T lymphocyte by MTT assay, killing activity of natural killer cells by LDH-release method, contents of CD3+, CD4+, CD8+, CD19+on flow cytometry, contents of serum hemolysin by HC50assay, serum contents of IL-1, IL-4and IFN-y by ELISA.
Results:1, Every dose group in Yupingfengsan and Fuqizhihan granule can improve innate immunity of Immunosuppressed mice in different degree. Compared with saline group, all indexes in cyclophosphamide group were statistically significant difference, besides, low dose group could improve killing activity of natural killer cells, phagocytosis index, serum IL-1β,in certain extent, but compared with saline group had statistically significant difference (P<0.05). Compared with cyclophosphamide group, all index except killing activity of natural killer cells and IL-1β had statistically difference.2, The influence of two compounds on adaptive immunity. CD19+,IL-4serum hemolysin, IFN-y, proliferation of T lymphocyte and contents of CD3+, CD4+, CD8+in cyclophosphamide group had statistically significant difference compared with saline group (P<0.05). low dose group could improve CD19+,IL-4serum hemolysin, IFN-y in some extent, but compared with saline group had statistically significant difference(P<0.05). Besides, compared with cyclophosphamide group low dose group could not improve CD19+,IL-4serum hemolysin, IFN-y (P>0.05), but high, middle dose group could improve adaptive immunity in some extent (P<0.05)
Conlusion:Both Yupingfengsan and Fuqizhihan granule could improve immunosuppressive mice immunity, the result revealed that their effect was similar. Both two compounds were superior in restoring cell immunity for immunosuppressive mice. Middle dose group had the best effect.
引文
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