加味四君子汤对脾气虚犬血清细胞因子水平、肠道免疫球蛋白水平以及脾脏组织形态及功能的影响

Effect of supplementary Sijunzi decoction on serum cytokine levels, intestinal immunoglobulin levels and spleen tissue morphology and function in Spleen-Qi deficiency canines

【目的】研究加味四君子汤对脾气虚犬免疫机能的影响.【方法】将24只1岁龄健康‘比格犬’随机均分为对照组、脾气虚组、自然恢复组和加味四君子汤组,用番泻叶药液复制犬脾气虚模型.第7 d造模成功,脾气虚组犬处死采样,对照组和自然恢复组犬饲喂基础日粮,加味四君子汤组犬在基础日粮中添加2%加味四君子汤.试验期14 d.第0、7、14天,采集血液,采用酶联免疫吸附法测定血清中干扰素(IFN)-γ、白细胞介素(IL)-2、IL-4、IL-6水平;在第14天,全部试验犬处死采样,计算脾脏指数,测定脾脏组织中促吞噬肽(Tuftsin)水平,观察脾脏显微和超微结构的变化;免疫组织化学(IHC)检测小肠各段分泌型免疫球蛋白A(sIgA)蛋白阳性表达水平,荧光定量PCR检测小肠各段免疫球蛋白A(IgA)、IgG、IgM mRNA相对表达量.【结果】与对照组犬相比,脾气虚犬血清细胞因子IL-4、IL-6水平显著升高(P<0.05),IFN-γ、IL-2水平显著降低(P<0.05);脾脏指数、脾脏Tuftsin水平、小肠sIgA蛋白阳性表达水平以及IgA、IgG、IgM mRNA的相对表达量均显著或极显著降低(P<0.05,P<0.01).经加味四君子汤治疗后,脾气虚犬上述指标和脾脏组织结构恢复正常.【结论】脾气虚犬免疫功能低于对照组犬,饲粮中添加加味四君子汤可以显著改善脾气虚症状,增强机体免疫机能.

【Objective】To study the effect of supplementary Sijunzi decoction(SJZD)on immune function of spleen qi deficiency canines.【Method】24 one-year-old healthy beagle canines were randomly divided into control group,spleen qi deficiency group,natural recovery group and supplementary SJZD group.The senna solution was used to replicate spleen qi deficiency model canines.The spleen qi deficiency model was established on 7 d,then the canines in spleen qi deficiency group were killed for collecting samples.The canines in the control group and the natural recovery group were fed a basal diet,the canines in supplementary SJZD group were fed a basal diet with 2% supplementary SJZD.The trial lasted for 14 d.The blood samples were taken at the 0,7 and 14 d,and the serum levels of IFN-γ,IL-2,IL-4,IL-6 were detected by enzyme linked immunosorbent assay(ELISA).On the 14 d,all the canines were killed for collecting samples,and the spleen index was calculated,the spleen Tuftsin level was detected,the microscopic and ultrastructural changes in spleen were observed.In small intestine,the protein positive expression level of sIgA was measured by immunohistochemistry(IHC),and the relative expression values of IgA,IgG and IgM mRNA were measured by fluorescent quantitative PCR.【Result】Compare with the control group canines,the levels of serum cytokines IL-4 and IL-6 in the spleen qi deficiency canines were significantly increased(P<0.05)and the levels of IFN-γand IL-2 were significantly decreased(P<0.05).The spleen index,spleen Tuftsin level and the protein positive expression level of sIgA and the relative expression values of IgA,IgG and IgM mRNA in small intestine were all significantly or extremely significantly decreased(P<0.05,P<0.01).After treatment with supplementary SJZD,the above indexes and spleen tissue structure of the spleen qi deficiency canines returned to normal.【Conclusion】The immune function of the spleen qi deficiency canines decreased than the control group canines,the basal diet with supplementary can significantly improve the symptoms of spleen qi deficiency and enhance the body's immune function.