黄芪多糖调节创伤应激小鼠免疫功能的研究

Study on effect of Astragalus polysaccharide for traumatic stress mice cell immunity

目的采用小鼠截肢应激模型,探讨创伤应激状态下小鼠免疫功能变化及黄芪多糖对应激状态下机体免疫功能影响。方法将50只BALBc小鼠随机分为5组正常对照组(A),每天腹腔注射生理盐水0.5ml只;应激对照组(B),截肢后每天腹腔注射生理盐水0.5ml只;应激+APS高(C)、中(D)、低(E)剂量组,截肢后分别给予APS1000mgkg、500mgkg、250mgkg,生理盐水稀释至0.5ml腹腔注射,每天1次。连续注射3d后,采用免疫组织化学技术检测CD4+、CD8+、cfos蛋白表达,原位杂交检测NFκBmRNA、IL10mRNA表达,计算机图像分析系统定量。结果与A组比较,创伤后72hB组小鼠胸腺、脾脏组织中NFκBmRNA、IL10mRNA表达水平均明显升高(P<0.01);CD4+、CD4+CD8+比值显著减少(P<0.01);cfos抗原表达明显多于正常(P<0.01)。与B组比较,C、D、E组小鼠胸腺、脾脏组织中NFκBmRNA、IL10mRNA的表达受抑(P<0.01);胸腺、脾脏组织中CD4+抗原水平、CD4+CD8+比值升高(P<0.01);胸腺、脾脏组织中cfos抗原水平降低(P<0.01)。与A组比较,C组胸腺、脾脏组织中NFκBmRNA、IL10mRNA的表达,CD4+抗原、CD8+抗原、cfos抗原分布差异无显著性(P>0.05)。结论创伤后小鼠细胞免疫功能明显紊乱;而黄芪多糖体内应用可有效恢复其免疫功能。

Objective To investigate the effects of traumatic stress on cell immunity of mice amputee model, and the influences of Astragalus polysaccharide (APS) on the cell immunity. Methods 50 BALB/c mice were divided randomly into 5 groups: group A (normal control group), group B (traumatic stress group), group C(traumatic stress + high dose APS group), group D(traumatic stress + middle dose APS group) and group E(traumatic stress+low dose APS group). The groups A and B were given 0.5ml of normal saline by interaperitoneal injection daily, and the groups C, D and E were given 1000mg/kg, 500mg/kg and 250mg/kg of APS by interaperitoneal injection daily for three days. CD4+, CD8+ and expression of c-fos protein were detected by immunohistochemisty method. Expression of NF-κB mRNA and IL-10 mRNA were detected by hybridization in situ. Their results were quantitated by HPIAS-1000 system. Results Compared with group A, in thymus and spleen tissues, the expression level of NF-κB mRNA, IL-10 mRNA and the distribution of c-fos antigen of group B were elevated significantly (P<0.01), CD4+ and CD4+/CD8+ were reduced(P<0.01). Compared with group B, the level of expression of NF-κB mRNA and IL-10 mRNA in thymus and spleen tissues were not only depressed by APS and the level of CD4+ antigen, CD4+/CD8+ ratio were elevated in groups C, D and E; but the level of c-fos antigen were reduced(P<0.01). The expression of NF-κB mRNA and IL-10 mRNA, CD4+ and CD4+/CD8+ and the distribution of c-fos antigen had no significant difference in both groups A and C (P>0.05). Conclusion The cell immunity function of mice were disordered after their trauma, APS may play a role in restoring cell immunity.