含红芪、黄芪的益气养血汤对SAMP8小鼠免疫衰老及p38MAPK表达的影响

Effects of Yiqi Yangxue Tang Containing Radix Astragali or Radix Hedyseri on Immunosenescence and p38MAPK Expression of SAMP8 Mice

目的:比较含红芪和含黄芪的益气养血汤对快速老化小鼠P8(SAMP8)小鼠免疫功能的影响,并探讨其可能的作用机制。方法:取昆明种青龄小鼠10只为青龄鼠组,SAMP8小鼠40只随机分为4组,分别为胸腺肽组、生理盐水组、益气养血红芪组和益气养血黄芪组。分别以生理盐水,胸腺肽每日4.6 mg·kg-1,含红芪、黄芪的益气养血汤每日6 g·kg-1剂量ig小鼠,每日1次,连续ig 15次,末次ig后2 h眼球摘除取血及脾脏。酶联免疫吸附测定(ELISA)检测其血清白细胞介素-2(IL-2),γ干扰素(IFN-γ)含量,流式细胞术检测脾脏淋巴细胞表面CD3,CD4,CD8,CD28及CD152分子的表达情况,实时荧光定量聚合酶链式反应(Real-time PCR)检测p38 MAPK mRNA的表达情况,蛋白质免疫印迹(Western blot)及免疫组化法检测其p38MAPK蛋白表达。结果:与青龄小鼠比较,生理盐水组SAMP8小鼠IL-2,IFN-γ含量降低,CD8,CD28分子表达降低,CD4分子表达及CD28CD152分子双阳性表达增高,p38 MAPK mRNA及蛋白表达降低(P<0.05)。与生理盐水组比较,胸腺肽组及中药组均能提高SAMP8小鼠血清中IL-2,IFN-γ含量,胸腺肽组含量高于中药组,红芪组IL-2含量高于黄芪组;胸腺肽组及中药组均能提高CD28分子表达,降低CD28CD152双阳性分子表达,胸腺肽组及黄芪组双阳性表达率低于红芪组;中药组能提高SAMP8小鼠脾淋巴细胞表面CD8分子表达,且黄芪组CD8表达高于红芪组;胸腺肽组及黄芪组能降低CD4分子表达;胸腺肽组及中药组均能提高p38 MAPK mRNA表达,且胸腺肽组mRNA表达高于中药组,黄芪组高于红芪组;胸腺肽组及中药组均能提高p38MAPK蛋白含量及蛋白阳性率,且胸腺肽组优于中药组(P<0.05)。结论:含红芪、黄芪的益气养血汤均能改善SAMP8小鼠的免疫衰老现象,增加p38MAPK的表达。

Objective： To compare the effects of Yiqi Yangxue Tang containing Radix Astragali or Radix Hedyseri on the immune function of Senescence accelerated mouse P8（ SAMP8） mice. Method： Ten Kunming mice were taken as normal group. 40 SAMP8 mice were randomly divided into normal saline group,thymosins group,Yiqi Yangxue Astragali Radix group and Yiqi Yangxue Hedyseri Radix group. The normal saline and thymosins were intragastrically administrated at 4. 6 mg·kg- 1daily,and the mice in the Yiqi Yangxue Astragali Radix group and the Yiqi Yangxue Hedyseri Radix group received treatment at the dose of 6 g·kg- 1by ig,once a day for fifteen times. Eyeball was taken for blood 2 h after the last ig administration; then spleen was also taken.The contents of interleukins-2（ IL-2） and γ-interferon（ IFN-γ） in the serum were detected by ELISA; the expression levels of CD3,CD4,CD8,CD28 and CD152 on splenic lymphocyte surface were detected by using flow cytometry; mRNA and protein expression levels of p38 MAPK were detected by Real-time PCR and Western blot respectively. Result： As compared with the normal group,the contents of IL-2 and IFN-γ were reduced; the expression of CD8 and CD28 molecules were decreased; CD4 molecule expression and CD28CD152 molecular double positive expression were increased; p38 MAPK mRNA and protein expression levels were decreased in the normal saline group（ P〈0. 05）. As compared with the normal saline group,IL-2 and IFN-γ contents in the serum of SAMP8 mice were increased in thymosins group and Yiqi Yangxue groups; the contents in the thymosins group were higher than those in Yiqi Yangxue groups; the level of IL-2 in Hedyseri Radix group was higher than that in the Astragali Radix group; the expression level of CD28 was increased and the double positive expression level of CD28,CD152 was reduced in both thymosins group and Yiqi Yangxue groups; the double positive expression rate in the thymosins group and Astragali Radix group was lower than that in Hedyseri Radix group. Yiqi Yangxue groups increased the expression levels of CD8 molecules and the expression in Astragali Radix group was higher than that in Hedyseri Radix group. The expression level of CD4 was decreased in both thymosins group and Astragali Radix group. The p38 MAPK mRNA expression level was increased in both thymosins group and Yiqi Yangxue groups,and the level in thymosins group was higher than that in Yiqi Yangxue groups; the level in Astragali Radix group was higher than that in Hedyseri Radix. p38 MAPK protein content and protein positive rate were increased in both thymosins group and Yiqi Yangxue groups groups,and the effect in thymosins group was higher than that in Yiqi Yangxue groups（ P〈0. 05）. Conclusion： The Yiqi Yangxue Tang containing Astragali Radix or Hedyseri Radix could improve the immunosenescence performances of SAMP8 mice,which may be associated with up-regulating p38 MAPK expression levels.