针灸对溃疡性结肠炎大鼠Th1/Th2免疫平衡的影响

目的:探讨针灸对溃疡性结肠炎(UC)大鼠Th 1/Th 2免疫平衡的影响。方法:SD大鼠随机分为模型组、隔药灸组、电针组,每组10只,并另选10只正常大鼠作为对照组。采用免疫学方法并加局部刺激建立大鼠UC模型。隔药灸组、电针组选取"天枢"(双)和"气海"进行隔药灸或电针治疗,连续治疗14d。采用定量PCR法检测结肠组织中干扰素(INF)-γ、白介素(IL)-12、IL-4和IL-10mRNA表达水平;采用酶联免疫吸附法检测结肠组织中INF-γ、IL-12、IL-4和IL-10的含量;采用流式细胞术分析结肠组织细胞中CD4~+INF-γ~+/CD 4~+IL-4~+的比值。结果:与对照组比较,模型组大鼠结肠组织中INF-γ、IL-12mRNA和含量显著升高(P0.05),IL-4、IL-10mRNA和含量显著下降(P0.05);隔药灸组和电针组大鼠结肠组织INF-γ、IL-12mRNA和含量显著低于模型组(P0.05),IL-4、IL-10mRNA和含量显著高于模型组(P0.05)。模型组结肠组织中CD 4~+INF-γ~+/CD 4~+IL-4~+的比值显著高于对照组(P0.05);与模型组比较,隔药灸组和电针组CD4~+INF-γ~+/CD 4~+IL-4~+比值显著下降(P0.05)。隔药灸组和电针组的各项指标比较差异均无统计学意义(P0.05)。结论:隔药灸和电针均能显著改善UC模型大鼠炎性反应,通过下调结肠组织中INF-γ和IL-12水平,上调IL-4和IL-10水平从而保持Th 1/Th 2细胞间平衡,进而改善免疫功能。

Effect of Moxibustion and Electroacupuncture on Th 1/Th 2 Immune Balance in Rats with Experimental Colitis

Objective To observe the effect of moxibustion and electroacupuncture(EA)stimulation of"Tianshu"(ST 25)and"Qihai"(CV 6)on Th 1/Th 2immune balance in rats with experimental ulcerative colitis(UC).Methods The UC rats were randomly allocated to model group,moxibustion group and EA group(n=10in each group).Another 10 rats were used as the control group.The UC model was established by injection of fresh colonic mucosal tissue-made antigen emulsifier(purified mucosal supernate plus complete Freund's adjuvant,3.5mg on day one,and 7mg thereafter)into the bilateral toes,inguinal grooves,back,and the abdominal cavity on day 1,10,17 and 24,respectively.Medicinal paste(containing Cortex Cinnamomi,Flos Carthami,etc.)-separated moxibustion or EA(200Hz,2mA)was applied to ST 25 and CV 6for 20 min,once daily for 14 days.The mRNA expression levels of interferon-γ(INF-γ),IL-12,IL-4 and IL-10 in colon tissues were detected using quantitative RT-PCR and their content determined using enzyme linked immunosorbent assay(ELISA).The colon tissue cell ratios of CD 4~+INF-γ~+and CD 4~+IL-4~+were analyzed by flow cytometry.Results Compared with the control group,the mRNA and contents of colonic INF-γ and IL-12 and the ratio of CD 4~+INF-γ~+and CD 4~+IL-4~+were significantly higher in the model group(P<0.05),and the IL-4 and IL-10 mRNA expression and contents considerably lower after UC modeling(P<0.05).Following medicinal paste-separated moxibustion and EA interventions,the increased INF-γand IL-12 mRNA and contents and the increased ratio of CD 4~+INF-γ~+and CD 4~+IL-4~+,and the decreased IL-4and IL-10 mRNA and contents were reversed(P<0.05).No significant differences were found between the moxibustion and EA groups in the levels of all the indexes(P>0.05).Conclusion Both medicinal paste-separated moxibustion and EA interventions may regulate the balance between Th 1and Th 2cells in the colonic tissues of UC rats,suggesting an improvement of the immune function by down-regulating INF-γand IL-12 levels and up-regulating IL-4and IL-10 levels.