从细胞凋亡角度验证隔药灸对溃疡性结肠炎大鼠环氧合酶2及前列腺素E_2的调节作用

Regulatory effect of herbs-partition moxibustion on cycloxygenase and prostaglandin E_2 in ulcerative colitis rats at apoptosis directions

背景:研究证实,艾灸有可能通过细胞因子调节环氧合酶2及前列腺素E2表达、影响细胞凋亡。目的:实验拟观察环氧合酶2、Bcl-2、Bax的表达及前列腺素E2的含量与溃疡性结肠炎的关系,探讨灸疗的影响作用并比较隔药灸和温和灸治疗作用的差异。设计、时间及地点:随机对照动物实验,于2006-04/2007-03在上海中医药大学实验动物中心和国家中医药管理局针灸免疫实验室完成。材料:SPF级雄性SD大鼠40只,体质量(140±20)g,采用免疫学方法加局部刺激制备溃疡性结肠炎大鼠模型。方法:40只大鼠随机数字表法分为正常组、模型组、隔药灸组、温和灸组,每组10只。正常组:不做任何治疗。模型组:只作与各治疗组相同的抓取固定。温和灸组:选取天枢、气海穴悬灸,每次每穴灸10min。隔药灸组:选取天枢、气海穴隔药灸,每次每穴灸2壮。疗程:1次/d,共灸14次。主要观察指标:免疫组织化学法检测溃疡性结肠炎大鼠结肠组织中环氧合酶2、Bcl-2、Bax的表达;放射免疫法测定溃疡性结肠炎大鼠结肠组织中前列腺素E2的含量。结果:纳入SD大鼠40只,造模过程有1只大鼠死亡,解剖见结肠明显扩张,未见穿孔,39只进入结果分析。模型组大鼠结肠组织环氧合酶2、Bcl-2的表达及前列腺素E2的含量高于正常组(P<0.01);而Bax的表达低于正常大鼠(P<0.01)。经过灸治后,与模型组比较,隔药灸组和温和灸组环氧合酶2、Bcl-2的表达及前列腺素E2的含量均显著降低(P<0.01);隔药灸组Bax的表达上调(P<0.05)。结论:灸疗干预可调节溃疡性结肠炎大鼠结肠组织环氧合酶2、Bax、Bcl-2的表达并降低前列腺素E2的含量,隔药灸的作用优于温和灸。

BACKGROUND： Previous researches show that moxibustion can regulate the expression of cycloxygenase （COX-2） and prostaglandin E2 （PGE2） and affect cell apoptosis through cytokine. OBJECTIVE： The study observed the correlation of the expression of COX-2, Bax, Bcl-2, PGE2 content to ulcerative colitis, and explored the effects of the moxibustion, compared the differences between herbs-partition moxibustion and mild moxibustion. DESIGN, TIME AND SETTING： Randomized control animal experiments were performed at the Animal Experimental Center of Shanghai University of TCM and Laboratory of Acupuncture and Immunity of State Administration of Traditional Chinese Medicine from April 2006 to March 2007. MATERIALS： Total 40 male SD rats （SPF grade） with body weight of （140±20） g were selected to establish rat models of ulcerative colitis by immunology and local excitation. METHODS： Forty rats were randomly divided into a normal control group, a model group, a herbs-partition moxibustion group, a mild-warm moxibustion group and each group have 10 rats. Rat models in the normal control group did not do any therapy. Rat models in the model group did grip fixation as the therapy group. Rat models in the mild-warm moxibustion group were treated by suspense moxibustion at the Tianshu （ST 25） and Qihai （RN 6） acupoints for 10 minutes. Rat models in the herbs-partition moxibustion group were treated by herbs-partition moxibustion at the Tianshu （ST 25） and Qihai （RN 6） acupoints for 2 zhuang. All treatments were used once a day for 14 days. MAIN OUTCOME MEASURES; Expression of COX-2, Bax, Bcl-2 was detected by immunohistochemistry. PGE2 content in colon tissue of ulcerative colitis rats was measured by radioimmunity. RESULTS： Thirty-nine SD rats were included in the final analysis. One rat was dead during modeling. Dissection showed that the rat＇s colon was obviously expanded, but did not perforate. The expression of COX-2, Bcl-2 and the content of PGE2 in colon tissue were significantly higher compared with control rats （P 〈 0.01）. The Bax expression was significantly lower than normal rats （P 〈 0.01）. After moxibustion therapy, the expression of COX-2, Bcl-2 and the content of PGE2 in the herbs-partition moxibustion group and the mild-warm moxibustion group were reduced compared with the model group （P 〈 0.01）. The Bax expression in the herbs-partition moxibustion group was upregulated （P 〈 0.05）. CONCLUSION： The expression of COX-2, Bax, Bcl-2 and the content of PGE2 can be regulated by moxibustion therapy. The effect of herbs-partition moxibustion is superior to mild moxibustion.