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从细胞因子基因表达角度研究针灸治疗溃疡性结肠炎大鼠的机制

Study of the mechanisms of acupuncture and moxibustion treatment for ulcerative colitis rats in view of the gene expression of cytokines.

作者信息

Wu Huan-Gan, Zhou Li-Bin, Pan Ying-Ying, Huang Cheng, Chen Han-Ping, Shi Zheng, Hua Xue-Gui

出版信息

World J Gastroenterol. 1999 Dec;5(6):515-517. doi: 10.3748/wjg.v5.i6.515.

Abstract

AIM

To observe the effect of acupuncture and moxibustion on the expression of IL-1beta and IL-6 mRNA in ulcerative colitis rats.METHODS:The SD rat ulcerative colitis model was created by immunological method associated with local stimulation. Colonic mucosa was prepared from human fresh surgical colonic specimens, homogenized by adding appropriate amount of normal saline and centrifuged at 3000r/min. The supernatant was collected for measurement of protein conentration and then mixed with Freund adjuvant. This antigen fluid was first injected into the plantae of the model group rats, and then into their plantae, dorsa, inguina and abdominal cavities (noFreund adjuvant for the last injection) again on the 10th, 17th, 24th and 31st day. When a certain titer of serum anti colonic antibody was reached, 2% formalin and antigen fluid (no Freund adjuvant) were administered separately by enema. The ulcerative colitis rat model was thus set up. The animals were randomly divided into four groups: model control group (MC, n = 8), electro acupuncture group (EA, n = 8), herbs partition moxibustion group (HPM 8), normal control group (NC,n = 8). HPM: Moxa cones made of refined mugwort floss were placed on the medicinal pad (medicinal pad dispensing: Radix Aconiti praeparata, cortex Cinnamomi, etc) for Qihai (RN 6) and Tianshu (ST 25, bilateral) and ignited. Two moxa cones were used for each acupoint once a day and 14 times in all. EA: Tianshu (bilateral) and Qihai were stimulated by the intermittent pulse with 2Hz frequency, 4mA intensity for 20 minutes once a day and 14 times in all. After treatment, rats of all four groups were killed simultaneously. The spleen was separated and the distal colon was dissected. Total tissue RNA was isolated by the guanidinium thiocyanate phenol chloroform extraction method. RT-PCR technique was used to study the expression of IL-1 beta and IL-6 mRNA.RESULTS:IL-1 beta and IL-6 mRNAs were not detected in the spleen and colonic mucosa of the NC rats, whereas they were significantly expressed in that of the MC rats.IL-1 beta and IL-6 mRNAs were markedly lower in the EA and HPM rats than that in MC rats. There was no significant difference between the levels of IL-1 beta and IL-6 mRNAs in the EA and HPM rats. The expressions of IL-1 beta and IL-6 mRNAs were nearly the same in the spleen and colon of all groups.CONCLUSION:Acupuncture and moxibustion greatly inhibited the expression of IL-1 beta and IL-6 mRNA in the experimental ulcerative colitis rats.

摘要

目的

观察针灸对溃疡性结肠炎大鼠白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)mRNA表达的影响。方法:采用免疫法结合局部刺激建立SD大鼠溃疡性结肠炎模型。取人新鲜手术结肠标本制备结肠黏膜,加入适量生理盐水匀浆,3000r/min离心,收集上清液测定蛋白浓度,然后与弗氏佐剂混合。将该抗原液先注入模型组大鼠足底,于第10、17、24、31天再分别注入其足底、背部、腹股沟及腹腔(最后一次注射不加弗氏佐剂)。当血清抗结肠抗体达到一定滴度时,分别用2%福尔马林和抗原液(不加弗氏佐剂)灌肠,建立溃疡性结肠炎大鼠模型。将动物随机分为四组:模型对照组(MC,n = 8)、电针组(EA,n = 8)、隔药灸组(HPM,n = 8)、正常对照组(NC,n = 8)。隔药灸组:将精制艾绒制成的艾炷置于药垫(药垫配方:制附子、肉桂等)上,放置于气海穴(任脉6)和双侧天枢穴(足阳明胃经25),点燃。每穴每次用2壮,每日1次,共14次。电针组:以2Hz频率、4mA强度的疏密波刺激双侧天枢穴和气海穴,每次20分钟,每日1次,共14次。治疗后,同时处死四组大鼠,分离脾脏,解剖远端结肠。采用异硫氰酸胍-酚-氯仿抽提法提取组织总RNA。运用逆转录聚合酶链反应(RT-PCR)技术研究IL-1β和IL-6 mRNA的表达。结果:正常对照组大鼠脾脏和结肠黏膜未检测到IL-1β和IL-6 mRNA,而模型对照组大鼠显著表达。电针组和隔药灸组大鼠IL-1β和IL-6 mRNA明显低于模型对照组大鼠。电针组和隔药灸组大鼠IL-1β和IL-6 mRNA水平无显著差异。各组大鼠脾脏和结肠中IL-1β和IL-6 mRNA的表达基本相同。结论:针灸可显著抑制实验性溃疡性结肠炎大鼠IL-1β和IL-6 mRNA的表达。

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