灌肠方治疗溃疡性结肠炎的临床和实验研究
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摘要
溃疡性结肠炎是一种以直肠、结肠粘膜非特异性炎症、溃疡形成为主要特征的病变。临床主要表现为腹泻、里急后重、粘液脓血便、腹痛等。病程迁延,常反复发作。其病因和发病机制目前认为主要与免疫、遗传、环境等因素的相互作用有关。近年来,溃疡性结肠炎的发病率在我国有增高趋势,临床上多采用美沙拉嗪及柳氮磺胺吡啶治疗。
溃疡性结肠炎归属于中医学“痢疾”、“泄泻”、“便血”“滞下”、“肠风”等范畴。其病因病机包括感受外邪、劳倦内伤或饮食不洁、情志失调、脾肾阳虚等方面。中医治疗溃疡性结肠炎的方法主要有中药方剂内服、灌肠、中药加灌肠、中药栓剂及其它剂型局部给药、单味中药、中西药结合、针灸等。其疗效显著,无明显副作用,并且价廉,显示出中医药治疗本病的优越性和广阔的前景。中药灌肠是临床常用的治疗方法之一,灌肠可以使药物直达病所,药物利用率高,且副作用少,所以临床应用较多。
本课题历经文献研究、临床研究、实验研究三个阶段,从临床及实验研究两方面来研究灌肠方治疗溃疡性结肠炎的临床疗效及其作用机制,为中医药治疗溃疡性结肠炎提供思路。论文分为三个部分：第一部分：文献综述
1.古代中医对溃疡性结肠炎病因、病机的总结,并对其辨证论治做了系统总结。现代中医对溃疡性结肠炎病因病机的认识,对其各种治疗方法做了系统论述,为本课题的临床研究提供理论基础和依据。
2.系统论述了近年来现代医学对溃疡性结肠炎发病机理的研究进展,包括免疫因素、细胞凋亡、遗传、精神因素及环境因素的研究进展,为本课题的实验研究提供理论依据。
第二部分：灌肠方治疗溃疡性结肠炎的临床研究
目的：1.通过采用随机对照的临床试验研究,观察灌肠方对溃疡性结肠炎湿热内蕴证患者的临床疗效和安全性,为中医药治疗溃疡性结肠炎提供临床依据。2.观察灌肠方对溃疡性结肠炎患者血清D-二聚体、血小板计数和血小板平均体积的影响,探讨灌肠方发挥作用的途径,分析湿热内蕴证和血瘀证之间的联系。
方法：采用随机对照的临床试验方法,入组40例,治疗组20例,对照组20例。治疗组给予灌肠方水煎液100ml保留灌肠,每晚1次,每日一剂。对照组给予柳氮磺胺吡啶口服,500mg／次,4次／d。治疗3周后,观察临床疗效、不良反应以及治疗前后血清D-二聚体、血小板计数和血小板平均体积水平的变化。
结果：治疗组总有效率为90.00%,对照组总有效率为75.00%,治疗组疗效明显高于对照组(P<0.05)。治疗组患者血清D-二聚体和血小板计数水平降低、血小板平均体积水平升高较对照组显著(P<0.05)。治疗组共发生可能与药物相关的不良反应1例(5%),对照组发生可能与药物相关不良反应2例(10%),两组不良反应发生率无显著差别(P＞0.05)。
结论：灌肠方治疗溃疡性结肠炎疗效显著,可能是通过清热利湿以改善瘀血状态；降低血清D-二聚体和血小板计数水平以及升高血小板平均体积水平可能是其治疗溃疡性结肠炎的机制之一。
第三部分：灌肠方治疗溃疡性结肠炎的实验研究
实验一：灌肠方对溃疡性结肠炎大鼠一般情况及结肠粘膜形态的影响。
目的：观察灌肠方对三硝基苯磺酸诱导的溃疡性结肠炎大鼠一般情况及结肠粘膜大体、病理变化的影响。
方法：取3月龄SD大鼠60只,雌雄各半,随机分为6组(正常组,空白对照组,中药低、中、高剂量组和西药美沙拉嗪对照组),每组10只。采用三硝基苯磺酸诱导法制作溃疡性结肠炎模型,造模后第3d开始给予灌胃给药,中药治疗组分别给予低、中、高剂量灌肠方水煎液,西药对照组给予美沙拉嗪,正常组及空白对照组给予等体积蒸馏水,每天1次,连续18天。观察大鼠一般情况、体重及治疗后结肠粘膜病理变化。
结果：造模后的大鼠出现粘液脓血便或稀便,自主活动减少,喜扎堆,食欲减退、倦怠懒动、反应迟缓。治疗18天后,上述情况改善,中药中、高剂量治疗组及西药对照组优于中药低剂量治疗组,各组大鼠的体重无明显差异。病理结果显示中药治疗组及西药对照组无粘膜层的坏死,仅有不同程度的炎症细胞浸润,优于空白对照组,中药中、高剂量组及西药对照组优于中药低剂量组。说明药物剂量与效果之间有一定相关性。
实验二：灌肠方对溃疡性结肠炎大鼠炎症细胞因子的影响。
目的：观察灌肠方对溃疡性结肠炎大鼠白介素-4和白介素-10的影响,探讨灌肠方在抗炎方面的作用。
方法：动物分组、造模及药物治疗同实验一,ELISA法检测各组大鼠血清白介素-4和白介素-10的水平。
结果：中药中、高剂量组和西药美沙拉嗪对照组之间无显著性差异,其余各组之间均有显著性差异(P<0.05)。空白对照组大鼠血清白介素-4和白介素-10的含量明显低于正常组,各中药治疗组及西药对照组均高于空白对照组。提示灌肠方可能是通过升高白介素-4和白介素-10的水平,纠正了促炎症性细胞因子和抗炎症性细胞因子之间的失衡,发挥治疗溃疡性结肠炎的作用。
实验三：灌肠方对溃疡性结肠炎大鼠过氧化物酶体增殖物激活受体的影响
目的：研究灌肠方对溃疡性结肠炎大鼠过氧化物酶体增殖物激活受体mRNA水平的影响。
方法：动物分组、造模及药物治疗同实验一,RT-PCR法检测各组大鼠结肠粘膜中过氧化物酶体增殖物激活受体mRNA的水平。
结果：空白对照组大鼠结肠粘膜中过氧化物酶体增殖物激活受体基因表达明显降低,与正常组比较有显著性差异(P<0.05)。中药中、高剂量组及西药对照组大鼠过氧化物酶体增殖物激活受体基因表达明显升高,与空白对照组比较,有显著性差异(P<0.05),而中药低剂量组与空白对照组比较,无显著性差异(P>0.05)。提示灌肠方可能是通过对溃疡性结肠炎大鼠低表达的过氧化物酶体增殖物激活受体的上调,抑制NF-κB的活化,进而抑制促炎因子等炎症相关因子表达,从而抑制炎症的发生及发展,类似于过氧化物酶体增殖物激活受体激动剂的作用,这可能是灌肠方发挥作用的机理之一
实验四：灌肠方对溃疡性结肠炎大鼠核因子-κB p65的影响
目的：研究灌肠方对溃疡性结肠炎大鼠核因子-κB p65mRNA水平的影响。
方法：动物分组、造模及药物治疗同实验一,RT-PCR法检测各组大鼠结肠粘膜中核因子-κB p65 mRNA的水平。
结果：空白对照组大鼠结肠组织中核因子-K B p65基因表达明显升高,与正常组比较有显著性差异(P<0.05)。中药低、中、高剂量组及西药对照组大鼠核因子-K B p65基因表达与空白对照组比较明显降低,有显著性差异(P<0.05),而中药低剂量组与中药中、高剂量组比较明显增高,有显著性差异(P<0.05)。提示灌肠方可能是通过对溃疡性结肠炎大鼠过高表达的核因子-K B p65的下调,抑制炎症因子的活化,减轻上皮细胞的损伤,从而抑制炎症的发生及发展。
实验五：灌肠方对溃疡性结肠炎大鼠Toll-like受体2的影响
目的：研究灌肠方对溃疡性结肠炎大鼠Toll-like受体2 mRNA水平的影响。
方法：动物分组、造模及药物治疗同实验一,RT-PCR法检测各组大鼠结肠粘膜中Toll-like受体2 mRNA的水平。
结果：空白对照组大鼠结肠组织中Toll-like受体2基因表达明显升高,与正常组比较有显著性差异(P<0.05)。中药低、中、高剂量组及西药对照组大鼠Toll-like受体2基因表达与空白对照组比较明显减低,有显著性差异(P<0.05),而中药低剂量组明显高于中药中、高剂量组,有显著性差异(P<0.05)。提示灌肠方对溃疡性结肠炎大鼠过高表达的Toll-like受体2有下调作用,阻止核因子-κB的激活,阻止多种致病因子的跨膜信号转导,减少致炎因子的释放,抑制过度的免疫反应,发挥治疗溃疡性结肠炎的作用。
结论：灌肠方发挥作用的机制可能是：1、通过升高白介素-4和白介素-10的水平,抑制促炎因子的合成,增加抗炎因子的大量合成,纠正了促炎症性细胞因子和抗炎症性细胞因子之间的失衡；抑制淋巴细胞介导的宿主自发免疫反应,阻止炎症的发生。2、上调过氧化物酶体增殖物激活受体的水平,抑制核因子-κB的活化和Toll-like受体2的表达,抑制促炎因子等炎症性相关因子的表达和活化,阻止多种致病因子的跨膜信号转导,减轻上皮细胞的损伤,抑制过度的免疫反应。
Objective
Ulcerative colitis is a disease of digestive tract characterised by chronic inflammation and ulceration of colonic mucous membrane. The inflammation of colon is chronic and nonspecific. The clinical manifestation of UC comprise mucus, pus and blood stool, diarrhea, abdominal pain, tenesmus and so on. UC usually last a very long period and do easy to relapse. The pathogenesis of ulcerative colitis is not very clear. Immune factors, environment, microorganism, heredity may have relationship with it. Among all of above, immune factors have been seriously concerned. At present, the incidence of ulcerative colitis is more and more rising. Sulfasalazine and mesalazine slow release tablets are primarily healing medicine in western medical science. In active stage of ulcerative colitis, these medicine can quickly control the symptoms. The clinical remission rate is high. But the disease is easily recur after drug withdrawal. In addition, the side effects of western medicine restrict its clinical using. Traditional Chinese Medicine shows its superiority in improving quality of life, lowering recurrence rate and lower spend. Guanchangfang is a proved recipe of guang'an men hospital, china academy of Chinese medical sciences. The curative effect of guanchangfang is positive which is proved in last curing patient. Based on the theory of Traditional Chinese Medicine, the effect of guanchangfang are dispelling dampness, heating-clearing and detoxicating, spasmolysis and alleviating pain. The topic evaluate its therapeutic effect. Besides, it investigates the mechanism of restraining the reaction of inflammatory injury, promoting the healing of colonic mucous membrane in the use of animal experiment. We analyze the mechanism of guanchangfang in order to offer an objective method and medicine to cure ulcerative colitis.
Methods
The clinical research study was strictly designed under correlated standard. The random positive control method was adopted, on the basis of diagnosis criterion established on Inflammatory Bowel Disease Seminar at chengdu in 2000 year.40 cases of ulcerative colitis are selected from out-patient clinic (OPD) of Qilu hospital of Shandong university. Treated group (20 cases) were received guanchangfang(coptis chinensis 10g, forsythia suspense 12g, jasmine fruit 10g, ledebourellae 12g, paeoniaealbe 12g, glycyrrhizae 10g). Control group were received sulfasalazine (2g/d). Both treatment course was 3 weeks. Mainly observed items:1. Therapeutic effect, including clinical general effect, change of Traditional Chinese Medicine syndrome.2. The serum levels of D-dimer (D-D)、Platelet (PLT) and Mean Platelet Volume (MPV).
In animal experiment, the SD rats were randomly divided into 6 groups:normal group, model group, treated group (low dose), treated group (middle dose) treated group (high dose) and mesalazine group. The last 5 groups are UC rats induced by 2,4,6—trinitrobenzenesulfonic Acid(TNBS)/ethanol mixture. The treatment time is 18 days, then we took serum and colon tissues from the rats. Mainly observed items:1. The effects of the general behaviors, living state and the weight of the rats.2. The pathological change of colon of rats both gross and microscopic observations.3. Serum level of Interleukin-4(IL-4), Interleukin-10(IL-10).4. The mRNA expression of peroxisome proliferators activated receptorγ(PPARγ), nuclear factor kB p 65(NF-kB p 65), Toll-Like Receptors 2(TLR 2)from colon mucous membrane were detected by reverse transcription PCR (RT-PCR).
Results
1. Evaluation of therapeutic effect of clinical research:
(1) Clinical therapeutic effect of treated group was better than control group. The total effective rate of treated group was 90.00% and control group was 75.00%. The statistic significance was difference. Treated group was obviously better than control group (P<0.05).
(2) The serum D-D and PLT of treated group were lower than control group. The statistic significance was difference (P<0.05). The serum MPV of treated group was remarkablely higher control group. The statistic significance was difference (P<0.05).
(3) Adverse effect rating of treated group was 5% and that of control group was 10%. The statistic significance was not difference (P>0.05).
2. Animal experiment research:
(1) On the aspect of the general behaviors, glossiness of coat and improving the living state, treated group and mesalazine group were better than model group. Besides, treated group (low dose) was worse than other treated group and mesalazine. Maybe the effect of medicine was concerned with the dose of medicine. The weight of all the rats was not significant difference. In addition, on the aspect of pathological change of colon of rats both gross and microscopic observations, treated group and mesalazine group were better than model group. Treated group (middle and high dose) and mesalazine group were better than treated group (low dose)
(2)Serum IL-4 of all groups was lower than normal group (P<0.05). Serum IL-4 of treated groups and mesalazine group were higher than model group (P<0.05) Treated group (middle and high dose) and mesalazine group were higher than treated group (low dose) (P<0.05). Treated group (middle and high dose) and mesalazine group were not significant difference(P>0.05).
(3) Serum IL-10 of all groups was lower than normal group (P<0.05). Serum IL-10 of treated groups and mesalazine group were higher than model group (P<0.05) Treated group (middle and high dose) and mesalazine group were higher than treated group (low dose) (P<0.05). Treated group (middle and high dose) and mesalazine group were not significant difference(P>0.05). Guanchangfang could increase the level of IL-4 which could improve the inflammation of colon.
(4) The colon mucous peroxisome proliferators—activated receptorγmRNA of normal group was significantly higher than other groups (P<0.05). The colon mucous peroxisome proliferators—activated receptorγmRNA of treated group (middle and high dose) and mesalazine group were higher than model group and treated group (low dose) (P<0.05). The colon mucous peroxisome proliferators—activated receptorγmRNA of treated group (low dose) was similar with model group (P>0.05). Maybe the effect of guanchangfang is elevation to the level of peroxisome proliferators—activated receptorγmRNA which could inhibit the activation of NF-κB. So the express of proinflammatory factor was interrupted. The development and growth of inflammation was prevented. The role of guanchangfang was similar with excitomotor of peroxisome proliferators activated receptorγ.
(5) The colon mucous nuclear factor k B p65 mRNA of model group, treated group and mesalazine group were significantly higher than normal groups (P<0.05) That of treated group and mesalazine group were lower model group (P<0.05) In addition, the colon mucous nuclear factorκB p65 mRNA of treated group (low dose) was higher than treated group (high and middle dose). Guanchangfang could cut down the level of nuclear factorκB p65 mRNA which could prevent the activation of proinflammatory factor. The deterioration of endothelial cell was improved and some ulcerative colitis was cured.
(6) The colon mucous Toll-Like Receptors 2 mRNA of model group, treated group and mesalazine group were significantly higher than normal groups (P<0.05) That of treated group and mesalazine group were lower model group (P<0.05) In addition, the colon mucous Toll-Like Receptors 2 mRNA of treated group (low dose) was higher than treated group (high and middle dose). Guanchangfang probably could control the gene expression of Toll-Like Receptors 2 and down-regulate the activation of nuclear factorκB which could block the signaling pathway of proinflammatory cytokines. The excessive immune response was influenced that maked endothelial cells restored. In addition, guanchangfang produce a marked effect in a dose-dependent manner. Above routes could be the mechanism that guanchangfang could improve the ulcerative colitis.
Conclusion
Guanchangfang is effective to treat ulcerative colitis whose type of syndrome is endoretention of damp heat by using the methods of clearing away heat evil, promoting diuresis and spasmolysis. The therapeutic effect is obviously better than SASP. It shows its superiority Compared with western medicine and has not obvious ill-effect. It can lower the serum content of D-dimer and platelet, and can increase mean platelet volume in sera, which may be an important pathway in clinical trail. The following is the mechanism of guanchangfang:1. Raising the expression of IL-4 and IL-10, preventing the aggregation of inflamed cell, helping the balance between proinflammatory factor and anti-inflammatory factor, restrainting the idiopathetic immune reaction.2. Raising the expression of colon mucous peroxisome proliferators—activated receptorγ, preventing nuclear factor k B and toll-Like receptors 2 so as to block the signaling pathway of proinflammatory cytokines, cut down the condition of local immunity, enhance mucosa epithelial cell proliferation and promote the repair of injuried mucous membrane.
溃疡性结肠炎归属于中医学“痢疾”、“泄泻”、“便血”“滞下”、“肠风”等范畴。其病因病机包括感受外邪、劳倦内伤或饮食不洁、情志失调、脾肾阳虚等方面。中医治疗溃疡性结肠炎的方法主要有中药方剂内服、灌肠、中药加灌肠、中药栓剂及其它剂型局部给药、单味中药、中西药结合、针灸等。其疗效显著,无明显副作用,并且价廉,显示出中医药治疗本病的优越性和广阔的前景。中药灌肠是临床常用的治疗方法之一,灌肠可以使药物直达病所,药物利用率高,且副作用少,所以临床应用较多。
本课题历经文献研究、临床研究、实验研究三个阶段,从临床及实验研究两方面来研究灌肠方治疗溃疡性结肠炎的临床疗效及其作用机制,为中医药治疗溃疡性结肠炎提供思路。论文分为三个部分：第一部分：文献综述
1.古代中医对溃疡性结肠炎病因、病机的总结,并对其辨证论治做了系统总结。现代中医对溃疡性结肠炎病因病机的认识,对其各种治疗方法做了系统论述,为本课题的临床研究提供理论基础和依据。
2.系统论述了近年来现代医学对溃疡性结肠炎发病机理的研究进展,包括免疫因素、细胞凋亡、遗传、精神因素及环境因素的研究进展,为本课题的实验研究提供理论依据。
第二部分：灌肠方治疗溃疡性结肠炎的临床研究
目的：1.通过采用随机对照的临床试验研究,观察灌肠方对溃疡性结肠炎湿热内蕴证患者的临床疗效和安全性,为中医药治疗溃疡性结肠炎提供临床依据。2.观察灌肠方对溃疡性结肠炎患者血清D-二聚体、血小板计数和血小板平均体积的影响,探讨灌肠方发挥作用的途径,分析湿热内蕴证和血瘀证之间的联系。
方法：采用随机对照的临床试验方法,入组40例,治疗组20例,对照组20例。治疗组给予灌肠方水煎液100ml保留灌肠,每晚1次,每日一剂。对照组给予柳氮磺胺吡啶口服,500mg／次,4次／d。治疗3周后,观察临床疗效、不良反应以及治疗前后血清D-二聚体、血小板计数和血小板平均体积水平的变化。
结果：治疗组总有效率为90.00%,对照组总有效率为75.00%,治疗组疗效明显高于对照组(P<0.05)。治疗组患者血清D-二聚体和血小板计数水平降低、血小板平均体积水平升高较对照组显著(P<0.05)。治疗组共发生可能与药物相关的不良反应1例(5%),对照组发生可能与药物相关不良反应2例(10%),两组不良反应发生率无显著差别(P＞0.05)。
结论：灌肠方治疗溃疡性结肠炎疗效显著,可能是通过清热利湿以改善瘀血状态；降低血清D-二聚体和血小板计数水平以及升高血小板平均体积水平可能是其治疗溃疡性结肠炎的机制之一。
第三部分：灌肠方治疗溃疡性结肠炎的实验研究
实验一：灌肠方对溃疡性结肠炎大鼠一般情况及结肠粘膜形态的影响。
目的：观察灌肠方对三硝基苯磺酸诱导的溃疡性结肠炎大鼠一般情况及结肠粘膜大体、病理变化的影响。
方法：取3月龄SD大鼠60只,雌雄各半,随机分为6组(正常组,空白对照组,中药低、中、高剂量组和西药美沙拉嗪对照组),每组10只。采用三硝基苯磺酸诱导法制作溃疡性结肠炎模型,造模后第3d开始给予灌胃给药,中药治疗组分别给予低、中、高剂量灌肠方水煎液,西药对照组给予美沙拉嗪,正常组及空白对照组给予等体积蒸馏水,每天1次,连续18天。观察大鼠一般情况、体重及治疗后结肠粘膜病理变化。
结果：造模后的大鼠出现粘液脓血便或稀便,自主活动减少,喜扎堆,食欲减退、倦怠懒动、反应迟缓。治疗18天后,上述情况改善,中药中、高剂量治疗组及西药对照组优于中药低剂量治疗组,各组大鼠的体重无明显差异。病理结果显示中药治疗组及西药对照组无粘膜层的坏死,仅有不同程度的炎症细胞浸润,优于空白对照组,中药中、高剂量组及西药对照组优于中药低剂量组。说明药物剂量与效果之间有一定相关性。
实验二：灌肠方对溃疡性结肠炎大鼠炎症细胞因子的影响。
目的：观察灌肠方对溃疡性结肠炎大鼠白介素-4和白介素-10的影响,探讨灌肠方在抗炎方面的作用。
方法：动物分组、造模及药物治疗同实验一,ELISA法检测各组大鼠血清白介素-4和白介素-10的水平。
结果：中药中、高剂量组和西药美沙拉嗪对照组之间无显著性差异,其余各组之间均有显著性差异(P<0.05)。空白对照组大鼠血清白介素-4和白介素-10的含量明显低于正常组,各中药治疗组及西药对照组均高于空白对照组。提示灌肠方可能是通过升高白介素-4和白介素-10的水平,纠正了促炎症性细胞因子和抗炎症性细胞因子之间的失衡,发挥治疗溃疡性结肠炎的作用。
实验三：灌肠方对溃疡性结肠炎大鼠过氧化物酶体增殖物激活受体的影响
目的：研究灌肠方对溃疡性结肠炎大鼠过氧化物酶体增殖物激活受体mRNA水平的影响。
方法：动物分组、造模及药物治疗同实验一,RT-PCR法检测各组大鼠结肠粘膜中过氧化物酶体增殖物激活受体mRNA的水平。
结果：空白对照组大鼠结肠粘膜中过氧化物酶体增殖物激活受体基因表达明显降低,与正常组比较有显著性差异(P<0.05)。中药中、高剂量组及西药对照组大鼠过氧化物酶体增殖物激活受体基因表达明显升高,与空白对照组比较,有显著性差异(P<0.05),而中药低剂量组与空白对照组比较,无显著性差异(P>0.05)。提示灌肠方可能是通过对溃疡性结肠炎大鼠低表达的过氧化物酶体增殖物激活受体的上调,抑制NF-κB的活化,进而抑制促炎因子等炎症相关因子表达,从而抑制炎症的发生及发展,类似于过氧化物酶体增殖物激活受体激动剂的作用,这可能是灌肠方发挥作用的机理之一
实验四：灌肠方对溃疡性结肠炎大鼠核因子-κB p65的影响
目的：研究灌肠方对溃疡性结肠炎大鼠核因子-κB p65mRNA水平的影响。
方法：动物分组、造模及药物治疗同实验一,RT-PCR法检测各组大鼠结肠粘膜中核因子-κB p65 mRNA的水平。
结果：空白对照组大鼠结肠组织中核因子-K B p65基因表达明显升高,与正常组比较有显著性差异(P<0.05)。中药低、中、高剂量组及西药对照组大鼠核因子-K B p65基因表达与空白对照组比较明显降低,有显著性差异(P<0.05),而中药低剂量组与中药中、高剂量组比较明显增高,有显著性差异(P<0.05)。提示灌肠方可能是通过对溃疡性结肠炎大鼠过高表达的核因子-K B p65的下调,抑制炎症因子的活化,减轻上皮细胞的损伤,从而抑制炎症的发生及发展。
实验五：灌肠方对溃疡性结肠炎大鼠Toll-like受体2的影响
目的：研究灌肠方对溃疡性结肠炎大鼠Toll-like受体2 mRNA水平的影响。
方法：动物分组、造模及药物治疗同实验一,RT-PCR法检测各组大鼠结肠粘膜中Toll-like受体2 mRNA的水平。
结果：空白对照组大鼠结肠组织中Toll-like受体2基因表达明显升高,与正常组比较有显著性差异(P<0.05)。中药低、中、高剂量组及西药对照组大鼠Toll-like受体2基因表达与空白对照组比较明显减低,有显著性差异(P<0.05),而中药低剂量组明显高于中药中、高剂量组,有显著性差异(P<0.05)。提示灌肠方对溃疡性结肠炎大鼠过高表达的Toll-like受体2有下调作用,阻止核因子-κB的激活,阻止多种致病因子的跨膜信号转导,减少致炎因子的释放,抑制过度的免疫反应,发挥治疗溃疡性结肠炎的作用。
结论：灌肠方发挥作用的机制可能是：1、通过升高白介素-4和白介素-10的水平,抑制促炎因子的合成,增加抗炎因子的大量合成,纠正了促炎症性细胞因子和抗炎症性细胞因子之间的失衡；抑制淋巴细胞介导的宿主自发免疫反应,阻止炎症的发生。2、上调过氧化物酶体增殖物激活受体的水平,抑制核因子-κB的活化和Toll-like受体2的表达,抑制促炎因子等炎症性相关因子的表达和活化,阻止多种致病因子的跨膜信号转导,减轻上皮细胞的损伤,抑制过度的免疫反应。
Objective
Ulcerative colitis is a disease of digestive tract characterised by chronic inflammation and ulceration of colonic mucous membrane. The inflammation of colon is chronic and nonspecific. The clinical manifestation of UC comprise mucus, pus and blood stool, diarrhea, abdominal pain, tenesmus and so on. UC usually last a very long period and do easy to relapse. The pathogenesis of ulcerative colitis is not very clear. Immune factors, environment, microorganism, heredity may have relationship with it. Among all of above, immune factors have been seriously concerned. At present, the incidence of ulcerative colitis is more and more rising. Sulfasalazine and mesalazine slow release tablets are primarily healing medicine in western medical science. In active stage of ulcerative colitis, these medicine can quickly control the symptoms. The clinical remission rate is high. But the disease is easily recur after drug withdrawal. In addition, the side effects of western medicine restrict its clinical using. Traditional Chinese Medicine shows its superiority in improving quality of life, lowering recurrence rate and lower spend. Guanchangfang is a proved recipe of guang'an men hospital, china academy of Chinese medical sciences. The curative effect of guanchangfang is positive which is proved in last curing patient. Based on the theory of Traditional Chinese Medicine, the effect of guanchangfang are dispelling dampness, heating-clearing and detoxicating, spasmolysis and alleviating pain. The topic evaluate its therapeutic effect. Besides, it investigates the mechanism of restraining the reaction of inflammatory injury, promoting the healing of colonic mucous membrane in the use of animal experiment. We analyze the mechanism of guanchangfang in order to offer an objective method and medicine to cure ulcerative colitis.
Methods
The clinical research study was strictly designed under correlated standard. The random positive control method was adopted, on the basis of diagnosis criterion established on Inflammatory Bowel Disease Seminar at chengdu in 2000 year.40 cases of ulcerative colitis are selected from out-patient clinic (OPD) of Qilu hospital of Shandong university. Treated group (20 cases) were received guanchangfang(coptis chinensis 10g, forsythia suspense 12g, jasmine fruit 10g, ledebourellae 12g, paeoniaealbe 12g, glycyrrhizae 10g). Control group were received sulfasalazine (2g/d). Both treatment course was 3 weeks. Mainly observed items:1. Therapeutic effect, including clinical general effect, change of Traditional Chinese Medicine syndrome.2. The serum levels of D-dimer (D-D)、Platelet (PLT) and Mean Platelet Volume (MPV).
In animal experiment, the SD rats were randomly divided into 6 groups:normal group, model group, treated group (low dose), treated group (middle dose) treated group (high dose) and mesalazine group. The last 5 groups are UC rats induced by 2,4,6—trinitrobenzenesulfonic Acid(TNBS)/ethanol mixture. The treatment time is 18 days, then we took serum and colon tissues from the rats. Mainly observed items:1. The effects of the general behaviors, living state and the weight of the rats.2. The pathological change of colon of rats both gross and microscopic observations.3. Serum level of Interleukin-4(IL-4), Interleukin-10(IL-10).4. The mRNA expression of peroxisome proliferators activated receptorγ(PPARγ), nuclear factor kB p 65(NF-kB p 65), Toll-Like Receptors 2(TLR 2)from colon mucous membrane were detected by reverse transcription PCR (RT-PCR).
Results
1. Evaluation of therapeutic effect of clinical research:
(1) Clinical therapeutic effect of treated group was better than control group. The total effective rate of treated group was 90.00% and control group was 75.00%. The statistic significance was difference. Treated group was obviously better than control group (P<0.05).
(2) The serum D-D and PLT of treated group were lower than control group. The statistic significance was difference (P<0.05). The serum MPV of treated group was remarkablely higher control group. The statistic significance was difference (P<0.05).
(3) Adverse effect rating of treated group was 5% and that of control group was 10%. The statistic significance was not difference (P>0.05).
2. Animal experiment research:
(1) On the aspect of the general behaviors, glossiness of coat and improving the living state, treated group and mesalazine group were better than model group. Besides, treated group (low dose) was worse than other treated group and mesalazine. Maybe the effect of medicine was concerned with the dose of medicine. The weight of all the rats was not significant difference. In addition, on the aspect of pathological change of colon of rats both gross and microscopic observations, treated group and mesalazine group were better than model group. Treated group (middle and high dose) and mesalazine group were better than treated group (low dose)
(2)Serum IL-4 of all groups was lower than normal group (P<0.05). Serum IL-4 of treated groups and mesalazine group were higher than model group (P<0.05) Treated group (middle and high dose) and mesalazine group were higher than treated group (low dose) (P<0.05). Treated group (middle and high dose) and mesalazine group were not significant difference(P>0.05).
(3) Serum IL-10 of all groups was lower than normal group (P<0.05). Serum IL-10 of treated groups and mesalazine group were higher than model group (P<0.05) Treated group (middle and high dose) and mesalazine group were higher than treated group (low dose) (P<0.05). Treated group (middle and high dose) and mesalazine group were not significant difference(P>0.05). Guanchangfang could increase the level of IL-4 which could improve the inflammation of colon.
(4) The colon mucous peroxisome proliferators—activated receptorγmRNA of normal group was significantly higher than other groups (P<0.05). The colon mucous peroxisome proliferators—activated receptorγmRNA of treated group (middle and high dose) and mesalazine group were higher than model group and treated group (low dose) (P<0.05). The colon mucous peroxisome proliferators—activated receptorγmRNA of treated group (low dose) was similar with model group (P>0.05). Maybe the effect of guanchangfang is elevation to the level of peroxisome proliferators—activated receptorγmRNA which could inhibit the activation of NF-κB. So the express of proinflammatory factor was interrupted. The development and growth of inflammation was prevented. The role of guanchangfang was similar with excitomotor of peroxisome proliferators activated receptorγ.
(5) The colon mucous nuclear factor k B p65 mRNA of model group, treated group and mesalazine group were significantly higher than normal groups (P<0.05) That of treated group and mesalazine group were lower model group (P<0.05) In addition, the colon mucous nuclear factorκB p65 mRNA of treated group (low dose) was higher than treated group (high and middle dose). Guanchangfang could cut down the level of nuclear factorκB p65 mRNA which could prevent the activation of proinflammatory factor. The deterioration of endothelial cell was improved and some ulcerative colitis was cured.
(6) The colon mucous Toll-Like Receptors 2 mRNA of model group, treated group and mesalazine group were significantly higher than normal groups (P<0.05) That of treated group and mesalazine group were lower model group (P<0.05) In addition, the colon mucous Toll-Like Receptors 2 mRNA of treated group (low dose) was higher than treated group (high and middle dose). Guanchangfang probably could control the gene expression of Toll-Like Receptors 2 and down-regulate the activation of nuclear factorκB which could block the signaling pathway of proinflammatory cytokines. The excessive immune response was influenced that maked endothelial cells restored. In addition, guanchangfang produce a marked effect in a dose-dependent manner. Above routes could be the mechanism that guanchangfang could improve the ulcerative colitis.
Conclusion
Guanchangfang is effective to treat ulcerative colitis whose type of syndrome is endoretention of damp heat by using the methods of clearing away heat evil, promoting diuresis and spasmolysis. The therapeutic effect is obviously better than SASP. It shows its superiority Compared with western medicine and has not obvious ill-effect. It can lower the serum content of D-dimer and platelet, and can increase mean platelet volume in sera, which may be an important pathway in clinical trail. The following is the mechanism of guanchangfang:1. Raising the expression of IL-4 and IL-10, preventing the aggregation of inflamed cell, helping the balance between proinflammatory factor and anti-inflammatory factor, restrainting the idiopathetic immune reaction.2. Raising the expression of colon mucous peroxisome proliferators—activated receptorγ, preventing nuclear factor k B and toll-Like receptors 2 so as to block the signaling pathway of proinflammatory cytokines, cut down the condition of local immunity, enhance mucosa epithelial cell proliferation and promote the repair of injuried mucous membrane.
引文
[1]李军昌,张宏博,成晓娟,等.溃疡性结肠炎患者98例的中医证型.第四军医大学学报,2003,24(5)：462.
[2]刘桂兰,石少波,黄笃高,等.中医治疗溃疡性结肠炎的思路.湖南中医药导报,2001,7(2)：61.
[3]李惠霞.参苓白术散加减配合中药灌肠治疗脾虚湿困型溃疡性结肠炎47例.辽宁中医杂志,2008,35(6)：886.
[4]杨运高.慢性非特异性结肠炎从淤论治初探.陕西中医,1993,14(1)：23-24.
[5]王波,史继英,杨亚东.中医辨证治疗慢性非特异性溃疡性结肠炎238例.中国中医急症,2006,15(2)：202.
[6]范恒,邱明义,段雪云,等.理肠四方治疗溃疡性结肠炎病机探讨.贵阳中医学院学报,2004,26(4)：17.
[7]夏永良.李德新教授治疗慢性结肠炎经验.北京中医药大学学报：中医临床版,2003,10(2)：35-36.
[8]顾培青.路广晁治疗溃疡性结肠炎经验.山东中医杂志,2006,25(10)：709-710.
[9]迟莉丽.隗继武教授治疗溃疡性结肠炎的临证应用.中医药学刊,2005,5(5)：787-788.
[10]劳国平.中医辩证加化瘀药治疗慢性非特异性溃疡性结肠炎的疗效观察.广西医科 大学学报,2000,17(5)：889-890.
[11]梁郁贤,宋仓棉.辨证治疗慢性非特异性溃疡性结肠炎39例.实用医技杂志,2006,3(3)：427.
[12]李天和.中药治疗溃疡性结肠炎66例.新中医,2002,34(10)：68.
[13]杨珍宝,董顺芳.中药辨证结合抗菌药物治疗慢性溃疡性结肠炎.中国肛肠病杂志,1998,18(7)：22-23.
[14]王伟民.溃疡性结肠炎辨治经验.中国肛肠病杂志,1997,17(1)：41-42.
[15]蔡全意,马腾飞,蔡静宜.辨证治疗溃疡性结肠炎162例报道.甘肃中医,2006,19(5)：17-18.
[16]黄德亚.下消补三步治疗溃疡性结肠炎9例.实用中医药杂志,1998,14(2)：31-32.
[17]杨现彩,杜丽萍.中医辨证治疗溃疡性结肠炎100例疗效观察.云南中医中药杂志,2005,26(4)：11.
[18]朱凤玲.中药灌肠治疗慢性溃疡性结肠炎50例.实用中医内科杂志,2005,19(1)：71.
[19]肖海,李长虹.中药灌注治疗溃疡性结肠炎17例临床分析.中国中西医结合外科杂志,2003,9(3)：189.
[20]于明环.灌肠散保留灌肠治疗溃疡性结肠炎141例.实用中医内科杂志,2004,18(2)：9.
[21]吴连恩,杨建平.五倍子灌肠液治疗慢性溃疡性结肠炎60例.江苏中医,1999,20(6)：21.
[22]张素琴,王明星.白头翁汤保留灌肠治疗慢性结肠炎120例.河南中医,1995,15(3)：147.
[23]陈庆华.愈溃汤保留灌肠治疗溃疡性结肠炎.山西中医,2004,20(2)：43.
[24]高祥华,指导李春婷.中医保留灌肠治疗慢性非特异性疡性结肠炎32例分析.吉林中医药,2003,23(12)：21.
[25]张尊环.白头翁灌肠治疗慢性溃疡性结肠炎37例.江苏中医,1998,19(6)：25.
[26]王美兰.血竭保留灌肠治疗慢性溃疡性结肠炎12例.安徽中医临床杂志,1999,11(1)：9.
[27]李柏林,韩永强.复方甘草片混悬液保留治疗慢性溃疡性结肠炎疗效观察.现代中西医结合杂志,2004,13(14)：1859.
[28]李春琳.中药煎剂保留灌肠治疗慢性溃疡性结肠炎26例疗效观察.中国民族民间医药杂志,2002,(59)：338.
[29]李德和,余涛.辨证分型治疗慢性溃疡性结肠炎经验.河北中医,2007,29(3)：228.
[30]孙静,单景烨.中药保留灌肠法治疗慢性非特异性溃疡结肠炎30例.河北中医,2005,27(12)：933.
[31]王子成.秦艽汤保留灌肠治疗溃疡性结肠炎.浙江中西医结合杂志,2002,12(8)：503.
[32]张锋,渠桂玲.阳和汤保留灌肠治疗溃疡性结肠炎78例临床观察.中国社区医师,2002,(1)：37.
[33]徐新勇,刘建武,陈秋红.加味膈下逐瘀汤保留灌肠治疗慢性结肠炎24例.国医论坛,2004,19(5)：24.
[34]刘华宝.中西医结合治疗慢性非特异性溃疡性结肠炎30例.四川中医,2006,24(4)：46-47.
[35]倪宝琢.中西医结合治疗慢性非特异性溃疡性结肠炎疗效观察.河北中医,2006,28(6)：447.
[36]赵增虎,丁瑞亮,范青键,等.思密达复合液保留灌肠治疗溃疡性结肠炎62例.实用中医内科杂志,2002,16(7)：22.
[37]刘谦民,毕锋,周绍娟.中西药联合灌肠治疗溃疡性结肠炎.新消化病杂志,1995,3(2)：115.
[38]汤效群.中药内外合治溃疡性结肠炎36例.安徽中医临床杂志,1998,10(2)：70.
[39]张进取.锡类散加西药保留灌肠治疗溃疡性结肠炎64例.安徽中医临床杂志,1998,10(2)：72.
[40]曹友平.综合疗法治疗慢性非特异性溃疡性结肠炎43例.新中医,2005,37(3)：70.
[41]金小晶,张文博,史肃玉,等.中药治疗慢性溃疡性结肠炎63例疗效观察.时珍国医国药,2005,16(11)：1146-1147.
[42]麦世杰.中药内服配合灌肠治疗溃疡性结肠炎36例.广西医学,2008,30(6)：851.
[43]侯冬梅,王利平,梁燕.针灸并用治疗慢性结肠炎30例.中医外治杂志,2004,13(2)：43.
[44]李应超.温脐化浊散外敷神阙穴、命门穴治疗慢性非特异性溃疡性结肠炎49例.中医外治杂志,2003,12(5)：20.
[45]刘兴,孙月霞.温针灸治疗溃疡性结肠炎30例.新疆中医药,2002,20(5)：33.
[46]杨顺益.针灸治疗溃疡性结肠炎的临床观察.上海针灸杂志,2001,20(1)：17.
[47]殷英.针药并用治疗慢性溃疡性结肠炎.中国社区医师,2006,8(4)：44.
[48]方向明,刘维洲,肖永俭,等.艾灸治疗溃疡性结肠炎58例疗效观察.针刺研究,1999,24(1)：66.
[49]赵维杰.穴位埋线治疗溃疡性结肠炎的临床研究.中国中医药杂志,2005,3(11)：975.
[50]王洪伟,韩宏妮,贾丽君.水罐疗法加针刺治疗溃疡性结肠炎56例.中国针灸杂志,2005,25(1)14.
[51]锁堂,吴焕,施达仁.白介素与溃疡性结肠炎.世界华人消化杂志,2006,14(4)：405.
[52]Macdonald TT, Monteleone G, Pender S L. Recent developments in the immunology of inflammatory bowel disease. Scan J Immunol,2000,51(1):2.
[53]Gan H, Ouyang Q, Jia D, et al. Activation of nuclear factor-kappaB and its relationship with cytokine gene expression in colonic mucosa of ulcerative colitis patients. Zhonghua Neike Zazhi,2002,41:252-255.
[54]张涛,谢建群.清肠栓对溃疡性结肠炎大鼠血清白介素影响的研究.辽宁中医药大学学报,2006,8(4)：50.
[55]Haruia J, Kusugam I K, Kuroiwa A, et al. Phenotypic and functional analysis of lam in a propria mononuclear cills from colono-scopic biopsy specimens in patients with ulcerative colitis. Am gastroenterolotgy,1992,87(4):448.
[56]Stevens C, Walz G, Singaran C, et al. Tumor necrosis factoralapha,interleukin I beta, interlcukin-6 expression in inflammatory bowel disease. Dig Dis Sei,1992,37:818.
[57]Wang L, Walia B, Evans J, et al. IL-6 induces NF-kappa B activation in the intestinal epithelia. Immunol,2003,171:3194-3201.
[58]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学讨论.中国现代医学杂志,2003,13(16)：74.
[59]Kullberg MC, Jankovic D, Feng CG, et al. IL-23 plays a key role in Helicobacter hepaticus-induced T cell-dependent colitis. J Exp Med,2006, 30;203(11):2485-2494.
[60]Szlosarek PW, Balkwill FR. Tumour necrosis factor alpha:a potential target for the therapy of solid tumours. Lancet Oncol,2003,4:565-573.
[62]Ebert EC. Tumour necrosis factor-alpha enhances intraepithelial lymphocyte proliferation and migration. Gut,1998,42(5):650-655.
[61]Hibi T, Inoue N, Ogata H, et al. Introduction and overview:recent advances in the immunotherapy of inflammatory bowel disease. Gastroenterol,2003,38 (Suppl 15):36-42.
[62]Nielsen OH, Koppen T, Rudiger. Involvement of interleukirr-4 and 10 in inflammatory bowel disease. Dig Dis Sei,1996,41:1786.
[63]崔海宏,陈村龙,杨玉捷,等.溃疡性结肠炎患者肠黏膜Th1/Th2类细胞因子-mRNA的表达.世界华人消化杂志,2003,11：1524-1527.
[64]Erdman SE, Rao VP, Poutahidis T, et al. CD4(+) CD25(+) regulatory lymphocytes require interleukin 10 to interrupt colon carcinogenesis in mice. Cancer Res,2003,63:6042-6050.
[65]Schreiber S, Fedorak RN, Nielsen OH, et al. Safety and efficaty of recombinant human interleukin 10 in chronic active Crohn's disease Crohn's disease IL-10 Cooperative Study Group. Gastroeterology,2000,119(6):1461.
[66]Melgar S, YeungM M, BasA, et al. Over-expression of interleukin-10 in mucosal T cells of patientswith active ulcerative colitis. Clin Exp Immunol, 2003,134:127-137.
[67]Fedorak RN, Gangl A, Elson CO, et al. Reconbinant human interleukin-10 in the treatment of patients with mild to moderately active Crohn's disease. The Interlenkin-10 Inflammatory Bowel Disease Cooperative Study Group. Gastroennterology,2000,199(6):1473.
[68]Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration:the multistep paradigm. Cell 1994,76:301-314.
[69]江学良,权启镇,孙自秦,等.溃疡性结肠炎患者粘附因子的变化意义.世界华人消化杂志,1998,6(6)：54-55.
[70]Briskin MJ, McEvoy LM, Butcher EC. MAdCAM-1 has homology to immunoglobulin and mucin—like adhesion receptors and to IgAl. Nature 1993,363:461-464.
[71]Van Assche G, Rutgeerts P. Physiological basis for novel drug therapies used to treat the inflammatory bowel diseases. Immunology and therapeutic potential of antiadhesion molecule therapy in inflammatory bowel disease. Am J Physiol Gastrointest Liver Physiol 2005,288:G169—174.
[72]Kato s, Hokari R, Matsuzaki K, Iwai A, et al. Amelioration of murine experimental colitis by inhibition of mucosal addressin cell adhesion molecule-1. J Pharmacof Exp 2000,295:183-189.
[73]Halstensen T. Epithelial deposition of immunogolbulin Gr and activated complement Ic3b and Terminal complement complex I in ulcerative colitis. Gastroenterology,1990,98:1264.
[74]Freeman H, Roecd B, Devine D, et al. Prospective evaluation of antineutrophil autoantibodise in 500 consecutive patients with inflammatory bowel disease. Can J Gastroenterol,1997,11:203.
[75]邓长生,夏冰主编.炎症性肠病.北京：人民卫生出版社,1998,131-145.
[76]Mulden TPJ, Verspaget HW, Janssens AR, et al. Decrease in two intestinal copper/zinc containing proteins w ith antioxidant function in inflammatory bowel disease. Gut,1991,32(10):1146.
[77]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学探讨.中国现代医学杂志,2003,8(13)：74-79.
[78]Caplan MS, Hedlund E, Hill N et al.The role of endogenous nitric oxide and platelet-activating factor in hypoxia-induced intestinal injury in rats. Gastroenterology.1994,106(2):346.
[79]Swidsinski A, Ladhoff A, Pernthaler A, et al. Mucosal flora in inflammatory bowel disease. Gatroenterology,2002,122:44-54.
[80]Miller MJ, Angeles FM, Reuter BK, et al. Dietary antioxidants protect gut epithelial cells from oxidant induced apoptosis. BMC Complement Altern Med,2001,1:11.
[81]GuobinHe, QinOu yang, Dai yunChen. The Microvacular Thrombi of Colonic Tissuein Ulcerative Colitis. Dig Dis Sci,2007,52(9):2236-2240.
[82]Collins CE, Rampton DS. Platelet dysfunction:a new dimension in inflammatory bowel disease. Gut,1995,36(1):5.
[83]Collins CE, Rampton DS. Platelets in inflammatory bowel disease pathogenetic role and therapeutic implications. A liment Pharmacol Ther, 1997,11 (2):237.
[84]Brannigan AE,0'Connell PR, Hurley H, et al. Neutrophil apoptosis is delayed in patients with inflammatory bowel disease. Shock,2000,13 (5):361-366.
[85]Ina K, usugami K, Hosokawa T, et al. Increased mucosal production of granulocyte colony-stimulating factor is related to a delay in neutrophil apoptosis in Inflammatory Bowel disease. Gastroenterol Hepatol,2001,14(1):46.
[86]Suzuki A, Sugimura K, Ohtsuka K, et al. Fas/Fax ligand expression and characateristics of primed CD45RO+T cell in the inflamed mucosa of ulcerative colitis. Scand J Gastroenterol 2001,35(22):1278.
[87]Neurath M, Finotto S, Fuss I, et al. Regulation of T-cell apoptosis in inflamatory bowel disease:To die or not to die, that is the mucosal guestion. Trends Immunol,2001,22(1):21.
[88]Cummings JH, Pomare EW, Branch WJ, et al. Short chain fatty acids in human large intestine, portal hepatic and venous blood. Gut,1987,28:1221.
[89]Vandcrhoof JA, Park JHY, Mohammadpour H. Effects of dietary lipids in recovery from mucosal injury. Gastroenterology,1990,98:1226-1331.
[90]Den Hond E, Hide M, Evenepoel P, et al. In vivo butyrate metabolism and colitis permeability in extensive ulcerative colit is. Gast roentero logy, 1998,115:584.
[9l]Scheppach W, Sommer H, Kirchner T, Paganelli GM, Bartram P, Christl S, Richter F, Dusel G, Kasper H. Effect of butyrate enemas on the colonic mucosa in distal ulcerative colitis. Gastroenterology,1992,103:51-56.
[92]Madsen KL, Doyle JS, Jewell LD, et al. Lactobacillus species prevents colitis in interleuk in 10 gene-deficient mice. Gastroenterology 1999,116: 1107.
[1]中华医学会消化病学分会.对炎症性肠病诊断治疗规范的建议(2000年).现代实用医学,2003,15(10)：656-659.
[2]贾黎明,周春立.炎症性肠病的内镜检查.见：郑家驹主编.炎症性肠病基础与临床.北京：科学出版社,2001,195.
[3]Ardizzone S, Dolodo P, Ranzi T, et al. Mesalazine foam (salofalk foam) in the treatment of active distal ulcerative colitis:a comparative trial vs Salofalk enema. The SAF-3 study group. Ital J Gastroenterol Hepatol,1999, 31(8):677.
[4]贺国斌,欧阳钦.溃疡性结肠炎的血栓前状态与肝素治疗.国外医学·内科学分册,2000,(6)：21.
[5]Hatoum OA, Spinelli KS, Abu-Hajir M, et al. Mesenteric venous thrombosis in inflammatory bowel disease. J Clin Gastroenterol,2005,39(1):27.
[6]Greenberg CS, Dcvinc DV, Mc Grac KM, et al. Measurement of plasma fibrin D-diner levels with the use of a monoclonal antibody coupled latex beed. Am J Clin Ppathol,1987,87(1):94-99.
[7]Bjerregaard LT, Nederby NT, Fredholm L, et al. Hyperhomocysteinaemia. Coagulation pathway activation and thrombophilia in patients with inflammtory bowel disease. Scand J Gastroenterol,2002,37(1):62.
[8]Collins CE, Rampton DS. Platelet dysfunction:a new dimension in inflammatory bowel disease. Gut,1995,36(1):5.
[9]Collins CE, Rampton DS. Platelets in inflammatory bowel disease pathogenetic role and therapeutic implications. Aliment Pharmacol Ther,1997,11(2):237.
[10]陈隆典,杨英.炎症性肠病疾病活动程度与血小板参数的相关性分析.中华消化杂志,2006,26(11)：771.
[11]Kapsoritakis AN, Koukourakis MI, Sfiridaki A, et al. Mean platelet volume: a useful marker of inflammatory bowel disease activity. Am J Gastroenterol,2001, 96(3):776.
[1]Morris GP, Beck PL, Herridge MS, et al. Hapte-induced model of chronic inflammation and ulceration in the rat colon. Gastroenterology,1989,96(3):795-803.
[2]锁堂,吴焕,施达仁. 白介素与溃疡性结肠炎. 世界华人消化杂志,2006,14(4)：405.
[3]Nielsen OH, Koppen T, Rudiger. Involvement of interleukirr-4 and 10 in inflammatory bowel disease. Dig Dis Sei,1996,41:1786.
[4]崔海宏,陈村龙,杨玉捷,等.溃疡性结肠炎患者肠黏膜Thl/Th2类细胞因子-mRNA的表达.世界华人消化杂志,2003,11：1524-1527.
[5]Melgar S, YeungM M, BasA, et al. Over-expression of inter-leukin 10 in mucosal T cells of patientswith active ulcerative colitis. Clin Exp Immunol,2003,134: 127-137.
[6]Dreyer C, Krey G, Keller H, et al. Control of the peroxisomal beta-oxidation pathway by a novel family of nuclear hormone receptors. Cell,1992,68(5):879-887.
[7]Meithaeghe A, Fajas L, Gouilleux F, et al. A functional polymorphism in a STA5B site of human PPAR gamma3gene promoter affects height and lipid metabolism in a French population. Arterropscler Thromb Vasc Biol,2003,23:L284.
[8]Malamut G, Cabane C, Dubuquoy L, et al.No evidence for an involvement of the p38 and JNK mitogen-activated protein in inflammatory bowel diseases. Dig Dis Sci.2006,51:1443-1453.
[9]Sanchez-Hidalgo M, Martin AR, Villegas I, et al. Rosiglitazone, a PPAR gamma ligand, modulates signal transduction pathways during the development of acute TNBS-induced colitis in rats. Eur J Pharmacol,2007,562(3):247-258.
[10]Desreumaux P, Ernst 0, Geboes K, et al. Inflammatory alterations in mesenteric adipose tissue in Crohn's disease. Gastroenterology,1999,117(1):73-81.
[11]Dubuquoy L, Jansson EA, Deeb S, et al. Impaired expression of peroxisome proliferator-activated receptor gamma in ulcerative colitis. Gastroenterology, 2003,124(5):1265-1276.
[12]Desreumaux P, Dubuquoy L, Nutten S, et al. Attenuation of colon inflammation through activators of the retinoid X receptor(RXR)/peroxisome proliferator-activated receptor gamma(PPARgamma) heterodimer:A basis for new therapeutic strategies.J Exp Med,2001,193(7):827-838.
[13]梁红亮,欧阳钦.过氧化物酶体增生物激活受体-γ在小鼠噁唑酮结肠炎中作用的研究.胃肠病学,2007,12(2)：101-105.
[14]Sen R, Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell,1986,46(5):705-716.
[15]Chen YG, Zhang C, Chiang SK, et al. Increased nuclear factor-kappa B p65 immunoreactivity following retinal ischemia and reperfusion injury in mice. J Neurosci Res,2003,72(1):125-131.
[16]Zaninoni A, Imperiali FG, Pasquini C, et al. Cytokine modulation of nuclear factor2kappa B activity in B2chronic lymphocytic leukemia. Exp Hematol,2003,31 (3):185-190.
[17]Chen YM, Tu CJ, Hung KY, et al. Inhibition by pentoxifylline of TNF-alpha stimulated f ractalkine production in vascular smoot h muscle cells:evidence for mediation by NF2kappa B downregulation. Br J Pharmacol,2003,138 (5):950-958.
[18]Kis A, Yellon DM, Baxter GF. Role of nuclear factor-kappa B activation in acute ischaemia reperfusion injury in myocardium. Br J Pharmacol,2003,138 (5):894-900.
[19]Calder VL, Bondeson J, Brennan FM, et al. Antigen-specific T2 cell down regulation by human dendritic cells following blockade of NF-kappaB. Scand J Immunol,2003,57(3):261-270.
[20]Abreu, Maria T. Immunologic regulation of toll-like receptors in gut epithelium. Gastroenterol,2003,19(6):559-564.
[21]Ashwood P, Harvey R, Ver jee T, et al. Funtional interactions between mucosal IL-1, IL-ra and TGF-bete 1 in ulcerative colitis. Inflamm Res,2004,53(2):53-59.
[22]Sawa Y, Oshitani N, Adachi K, et al. Comprehensive analysis of intestinal cytokine messenger RNA profile by real-time quantitative polymerase chain reaction in patient s with inflammatory bowel disease. Int J Mol Med,2003,11 (2):175-179.
[23]甘华田,欧阳钦,贾道全,等.溃疡性结肠炎患者核因子-κB活化与细胞因子基因表达.中华内科杂志,2002,41：252-255.
[24]Hashimoto C, Hudson KL, Anderson KV. The Toll Gene of Drosophila, required for dorsal-ventral embryonic polarity, appears to encode a transmembrane protein. Cell,1988,52(2):269-279.
[25]Medzhitov R, Preston-Hurlburt P, Janeway CA, et al. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature, 1997,388 (6640):394-397.
[26]Netea MG, Van derMeer JW, Kullberg BJ. Recognition of pathogenic microorganisms by Toll-like receptors. Drugs Today (Bare),2006,42 (supp lA): 99-105.
[27]Canto E, Ricart E, MonfortD, et al. TNF alphap roduction to TLR2 ligands in active IBD patients. Clin Immunol,2006,119(2):156-165.
[28]Cario E, Podolsky DK. Differential alteration in intestinal epithelial cell expression of Toll-like receptor 3 (TLR3) and TLR4 in inflammatory bowel disasel Infect Immun,2000,68(12):7010-7017.
[29]Toiyama Y, Araki T, Yoshiyama S, et al. The expression patterns of Toll-like receptors in the ileal pouch mucosa of postoperative ulcerative colitis patientsl Surg Today,2006,36(3):287-290.
[30]Douglas R, McDonald, Riny Janssen, Raif Geha. Lessons learned from molecular defects in nuclear factor κ B dependent signaling. Microbes and infection,2006,8(4):1151-1156.
[31]Hanauer, Stephen B MD. Inflammatory bowel disease: epidemiology, pathogenesis, and therapeutic opportunities. Inflammatory Bowl Disease,2006,12 (supplement 1):S3-S9.
[32]Abreu MT, Vora P, Fature E, et al. Decreased expression of Toll-like receptor-4 and MD-2correlates with intestinal epithelial cell protection against dysregulated proinflammatory gene expression in response to bacterial lipopoly saccharide. J Immunol,2001,167(3):1609-1617.
[33]Paul N, Moynag H. TLR signalling and activation of IRFs:revisiting old friends from the NF-κB pathway. Trends in immunology,2005,26(9):469-476.
[34]Liu Y, Wang Y, Yamankuchi M, et al. Upregulation of Toll-like receptor 2 gene expression in macrophage response to peptidoglycan and high concentration of lipopolysaccharide is involved in NF-κB activation. Infection Immun,2001,69 (5):2788-2796.
[35]刘一品,李延青.核因子-κB的表达在溃疡性结肠炎发病机制中的意义.胃肠病 学,2006,11(2)：103-106.
1、分析了溃疡性结肠炎湿热内蕴和血瘀之间的关系,并以现代医学高凝状态的指标验证血瘀证的病理状态,求证灌肠方的作用机理。
2、动物实验从调整细胞因子间的平衡状态、抑制自身免疫反应方面,从蛋白、基因水平多角度的分析灌肠方的作用机理,开阔了视野,拓展了中医药治疗溃疡性结肠炎的思路。
3、溃疡性结肠炎模型制作中,我们采用了双猪尾管从肛门插入,根据管上的刻度,易于掌握插入的长度；此管为软管,易于操作,并且对大鼠结肠粘膜的机械损伤小,更符合溃结的病理生理模型。
[2]刘桂兰,石少波,黄笃高,等.中医治疗溃疡性结肠炎的思路.湖南中医药导报,2001,7(2)：61.
[3]李惠霞.参苓白术散加减配合中药灌肠治疗脾虚湿困型溃疡性结肠炎47例.辽宁中医杂志,2008,35(6)：886.
[4]杨运高.慢性非特异性结肠炎从淤论治初探.陕西中医,1993,14(1)：23-24.
[5]王波,史继英,杨亚东.中医辨证治疗慢性非特异性溃疡性结肠炎238例.中国中医急症,2006,15(2)：202.
[6]范恒,邱明义,段雪云,等.理肠四方治疗溃疡性结肠炎病机探讨.贵阳中医学院学报,2004,26(4)：17.
[7]夏永良.李德新教授治疗慢性结肠炎经验.北京中医药大学学报：中医临床版,2003,10(2)：35-36.
[8]顾培青.路广晁治疗溃疡性结肠炎经验.山东中医杂志,2006,25(10)：709-710.
[9]迟莉丽.隗继武教授治疗溃疡性结肠炎的临证应用.中医药学刊,2005,5(5)：787-788.
[10]劳国平.中医辩证加化瘀药治疗慢性非特异性溃疡性结肠炎的疗效观察.广西医科 大学学报,2000,17(5)：889-890.
[11]梁郁贤,宋仓棉.辨证治疗慢性非特异性溃疡性结肠炎39例.实用医技杂志,2006,3(3)：427.
[12]李天和.中药治疗溃疡性结肠炎66例.新中医,2002,34(10)：68.
[13]杨珍宝,董顺芳.中药辨证结合抗菌药物治疗慢性溃疡性结肠炎.中国肛肠病杂志,1998,18(7)：22-23.
[14]王伟民.溃疡性结肠炎辨治经验.中国肛肠病杂志,1997,17(1)：41-42.
[15]蔡全意,马腾飞,蔡静宜.辨证治疗溃疡性结肠炎162例报道.甘肃中医,2006,19(5)：17-18.
[16]黄德亚.下消补三步治疗溃疡性结肠炎9例.实用中医药杂志,1998,14(2)：31-32.
[17]杨现彩,杜丽萍.中医辨证治疗溃疡性结肠炎100例疗效观察.云南中医中药杂志,2005,26(4)：11.
[18]朱凤玲.中药灌肠治疗慢性溃疡性结肠炎50例.实用中医内科杂志,2005,19(1)：71.
[19]肖海,李长虹.中药灌注治疗溃疡性结肠炎17例临床分析.中国中西医结合外科杂志,2003,9(3)：189.
[20]于明环.灌肠散保留灌肠治疗溃疡性结肠炎141例.实用中医内科杂志,2004,18(2)：9.
[21]吴连恩,杨建平.五倍子灌肠液治疗慢性溃疡性结肠炎60例.江苏中医,1999,20(6)：21.
[22]张素琴,王明星.白头翁汤保留灌肠治疗慢性结肠炎120例.河南中医,1995,15(3)：147.
[23]陈庆华.愈溃汤保留灌肠治疗溃疡性结肠炎.山西中医,2004,20(2)：43.
[24]高祥华,指导李春婷.中医保留灌肠治疗慢性非特异性疡性结肠炎32例分析.吉林中医药,2003,23(12)：21.
[25]张尊环.白头翁灌肠治疗慢性溃疡性结肠炎37例.江苏中医,1998,19(6)：25.
[26]王美兰.血竭保留灌肠治疗慢性溃疡性结肠炎12例.安徽中医临床杂志,1999,11(1)：9.
[27]李柏林,韩永强.复方甘草片混悬液保留治疗慢性溃疡性结肠炎疗效观察.现代中西医结合杂志,2004,13(14)：1859.
[28]李春琳.中药煎剂保留灌肠治疗慢性溃疡性结肠炎26例疗效观察.中国民族民间医药杂志,2002,(59)：338.
[29]李德和,余涛.辨证分型治疗慢性溃疡性结肠炎经验.河北中医,2007,29(3)：228.
[30]孙静,单景烨.中药保留灌肠法治疗慢性非特异性溃疡结肠炎30例.河北中医,2005,27(12)：933.
[31]王子成.秦艽汤保留灌肠治疗溃疡性结肠炎.浙江中西医结合杂志,2002,12(8)：503.
[32]张锋,渠桂玲.阳和汤保留灌肠治疗溃疡性结肠炎78例临床观察.中国社区医师,2002,(1)：37.
[33]徐新勇,刘建武,陈秋红.加味膈下逐瘀汤保留灌肠治疗慢性结肠炎24例.国医论坛,2004,19(5)：24.
[34]刘华宝.中西医结合治疗慢性非特异性溃疡性结肠炎30例.四川中医,2006,24(4)：46-47.
[35]倪宝琢.中西医结合治疗慢性非特异性溃疡性结肠炎疗效观察.河北中医,2006,28(6)：447.
[36]赵增虎,丁瑞亮,范青键,等.思密达复合液保留灌肠治疗溃疡性结肠炎62例.实用中医内科杂志,2002,16(7)：22.
[37]刘谦民,毕锋,周绍娟.中西药联合灌肠治疗溃疡性结肠炎.新消化病杂志,1995,3(2)：115.
[38]汤效群.中药内外合治溃疡性结肠炎36例.安徽中医临床杂志,1998,10(2)：70.
[39]张进取.锡类散加西药保留灌肠治疗溃疡性结肠炎64例.安徽中医临床杂志,1998,10(2)：72.
[40]曹友平.综合疗法治疗慢性非特异性溃疡性结肠炎43例.新中医,2005,37(3)：70.
[41]金小晶,张文博,史肃玉,等.中药治疗慢性溃疡性结肠炎63例疗效观察.时珍国医国药,2005,16(11)：1146-1147.
[42]麦世杰.中药内服配合灌肠治疗溃疡性结肠炎36例.广西医学,2008,30(6)：851.
[43]侯冬梅,王利平,梁燕.针灸并用治疗慢性结肠炎30例.中医外治杂志,2004,13(2)：43.
[44]李应超.温脐化浊散外敷神阙穴、命门穴治疗慢性非特异性溃疡性结肠炎49例.中医外治杂志,2003,12(5)：20.
[45]刘兴,孙月霞.温针灸治疗溃疡性结肠炎30例.新疆中医药,2002,20(5)：33.
[46]杨顺益.针灸治疗溃疡性结肠炎的临床观察.上海针灸杂志,2001,20(1)：17.
[47]殷英.针药并用治疗慢性溃疡性结肠炎.中国社区医师,2006,8(4)：44.
[48]方向明,刘维洲,肖永俭,等.艾灸治疗溃疡性结肠炎58例疗效观察.针刺研究,1999,24(1)：66.
[49]赵维杰.穴位埋线治疗溃疡性结肠炎的临床研究.中国中医药杂志,2005,3(11)：975.
[50]王洪伟,韩宏妮,贾丽君.水罐疗法加针刺治疗溃疡性结肠炎56例.中国针灸杂志,2005,25(1)14.
[51]锁堂,吴焕,施达仁.白介素与溃疡性结肠炎.世界华人消化杂志,2006,14(4)：405.
[52]Macdonald TT, Monteleone G, Pender S L. Recent developments in the immunology of inflammatory bowel disease. Scan J Immunol,2000,51(1):2.
[53]Gan H, Ouyang Q, Jia D, et al. Activation of nuclear factor-kappaB and its relationship with cytokine gene expression in colonic mucosa of ulcerative colitis patients. Zhonghua Neike Zazhi,2002,41:252-255.
[54]张涛,谢建群.清肠栓对溃疡性结肠炎大鼠血清白介素影响的研究.辽宁中医药大学学报,2006,8(4)：50.
[55]Haruia J, Kusugam I K, Kuroiwa A, et al. Phenotypic and functional analysis of lam in a propria mononuclear cills from colono-scopic biopsy specimens in patients with ulcerative colitis. Am gastroenterolotgy,1992,87(4):448.
[56]Stevens C, Walz G, Singaran C, et al. Tumor necrosis factoralapha,interleukin I beta, interlcukin-6 expression in inflammatory bowel disease. Dig Dis Sei,1992,37:818.
[57]Wang L, Walia B, Evans J, et al. IL-6 induces NF-kappa B activation in the intestinal epithelia. Immunol,2003,171:3194-3201.
[58]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学讨论.中国现代医学杂志,2003,13(16)：74.
[59]Kullberg MC, Jankovic D, Feng CG, et al. IL-23 plays a key role in Helicobacter hepaticus-induced T cell-dependent colitis. J Exp Med,2006, 30;203(11):2485-2494.
[60]Szlosarek PW, Balkwill FR. Tumour necrosis factor alpha:a potential target for the therapy of solid tumours. Lancet Oncol,2003,4:565-573.
[62]Ebert EC. Tumour necrosis factor-alpha enhances intraepithelial lymphocyte proliferation and migration. Gut,1998,42(5):650-655.
[61]Hibi T, Inoue N, Ogata H, et al. Introduction and overview:recent advances in the immunotherapy of inflammatory bowel disease. Gastroenterol,2003,38 (Suppl 15):36-42.
[62]Nielsen OH, Koppen T, Rudiger. Involvement of interleukirr-4 and 10 in inflammatory bowel disease. Dig Dis Sei,1996,41:1786.
[63]崔海宏,陈村龙,杨玉捷,等.溃疡性结肠炎患者肠黏膜Th1/Th2类细胞因子-mRNA的表达.世界华人消化杂志,2003,11：1524-1527.
[64]Erdman SE, Rao VP, Poutahidis T, et al. CD4(+) CD25(+) regulatory lymphocytes require interleukin 10 to interrupt colon carcinogenesis in mice. Cancer Res,2003,63:6042-6050.
[65]Schreiber S, Fedorak RN, Nielsen OH, et al. Safety and efficaty of recombinant human interleukin 10 in chronic active Crohn's disease Crohn's disease IL-10 Cooperative Study Group. Gastroeterology,2000,119(6):1461.
[66]Melgar S, YeungM M, BasA, et al. Over-expression of interleukin-10 in mucosal T cells of patientswith active ulcerative colitis. Clin Exp Immunol, 2003,134:127-137.
[67]Fedorak RN, Gangl A, Elson CO, et al. Reconbinant human interleukin-10 in the treatment of patients with mild to moderately active Crohn's disease. The Interlenkin-10 Inflammatory Bowel Disease Cooperative Study Group. Gastroennterology,2000,199(6):1473.
[68]Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration:the multistep paradigm. Cell 1994,76:301-314.
[69]江学良,权启镇,孙自秦,等.溃疡性结肠炎患者粘附因子的变化意义.世界华人消化杂志,1998,6(6)：54-55.
[70]Briskin MJ, McEvoy LM, Butcher EC. MAdCAM-1 has homology to immunoglobulin and mucin—like adhesion receptors and to IgAl. Nature 1993,363:461-464.
[71]Van Assche G, Rutgeerts P. Physiological basis for novel drug therapies used to treat the inflammatory bowel diseases. Immunology and therapeutic potential of antiadhesion molecule therapy in inflammatory bowel disease. Am J Physiol Gastrointest Liver Physiol 2005,288:G169—174.
[72]Kato s, Hokari R, Matsuzaki K, Iwai A, et al. Amelioration of murine experimental colitis by inhibition of mucosal addressin cell adhesion molecule-1. J Pharmacof Exp 2000,295:183-189.
[73]Halstensen T. Epithelial deposition of immunogolbulin Gr and activated complement Ic3b and Terminal complement complex I in ulcerative colitis. Gastroenterology,1990,98:1264.
[74]Freeman H, Roecd B, Devine D, et al. Prospective evaluation of antineutrophil autoantibodise in 500 consecutive patients with inflammatory bowel disease. Can J Gastroenterol,1997,11:203.
[75]邓长生,夏冰主编.炎症性肠病.北京：人民卫生出版社,1998,131-145.
[76]Mulden TPJ, Verspaget HW, Janssens AR, et al. Decrease in two intestinal copper/zinc containing proteins w ith antioxidant function in inflammatory bowel disease. Gut,1991,32(10):1146.
[77]王伟宁,张熙纯,刘丽.活动期溃疡性结肠炎发病机制的免疫学探讨.中国现代医学杂志,2003,8(13)：74-79.
[78]Caplan MS, Hedlund E, Hill N et al.The role of endogenous nitric oxide and platelet-activating factor in hypoxia-induced intestinal injury in rats. Gastroenterology.1994,106(2):346.
[79]Swidsinski A, Ladhoff A, Pernthaler A, et al. Mucosal flora in inflammatory bowel disease. Gatroenterology,2002,122:44-54.
[80]Miller MJ, Angeles FM, Reuter BK, et al. Dietary antioxidants protect gut epithelial cells from oxidant induced apoptosis. BMC Complement Altern Med,2001,1:11.
[81]GuobinHe, QinOu yang, Dai yunChen. The Microvacular Thrombi of Colonic Tissuein Ulcerative Colitis. Dig Dis Sci,2007,52(9):2236-2240.
[82]Collins CE, Rampton DS. Platelet dysfunction:a new dimension in inflammatory bowel disease. Gut,1995,36(1):5.
[83]Collins CE, Rampton DS. Platelets in inflammatory bowel disease pathogenetic role and therapeutic implications. A liment Pharmacol Ther, 1997,11 (2):237.
[84]Brannigan AE,0'Connell PR, Hurley H, et al. Neutrophil apoptosis is delayed in patients with inflammatory bowel disease. Shock,2000,13 (5):361-366.
[85]Ina K, usugami K, Hosokawa T, et al. Increased mucosal production of granulocyte colony-stimulating factor is related to a delay in neutrophil apoptosis in Inflammatory Bowel disease. Gastroenterol Hepatol,2001,14(1):46.
[86]Suzuki A, Sugimura K, Ohtsuka K, et al. Fas/Fax ligand expression and characateristics of primed CD45RO+T cell in the inflamed mucosa of ulcerative colitis. Scand J Gastroenterol 2001,35(22):1278.
[87]Neurath M, Finotto S, Fuss I, et al. Regulation of T-cell apoptosis in inflamatory bowel disease:To die or not to die, that is the mucosal guestion. Trends Immunol,2001,22(1):21.
[88]Cummings JH, Pomare EW, Branch WJ, et al. Short chain fatty acids in human large intestine, portal hepatic and venous blood. Gut,1987,28:1221.
[89]Vandcrhoof JA, Park JHY, Mohammadpour H. Effects of dietary lipids in recovery from mucosal injury. Gastroenterology,1990,98:1226-1331.
[90]Den Hond E, Hide M, Evenepoel P, et al. In vivo butyrate metabolism and colitis permeability in extensive ulcerative colit is. Gast roentero logy, 1998,115:584.
[9l]Scheppach W, Sommer H, Kirchner T, Paganelli GM, Bartram P, Christl S, Richter F, Dusel G, Kasper H. Effect of butyrate enemas on the colonic mucosa in distal ulcerative colitis. Gastroenterology,1992,103:51-56.
[92]Madsen KL, Doyle JS, Jewell LD, et al. Lactobacillus species prevents colitis in interleuk in 10 gene-deficient mice. Gastroenterology 1999,116: 1107.
[1]中华医学会消化病学分会.对炎症性肠病诊断治疗规范的建议(2000年).现代实用医学,2003,15(10)：656-659.
[2]贾黎明,周春立.炎症性肠病的内镜检查.见：郑家驹主编.炎症性肠病基础与临床.北京：科学出版社,2001,195.
[3]Ardizzone S, Dolodo P, Ranzi T, et al. Mesalazine foam (salofalk foam) in the treatment of active distal ulcerative colitis:a comparative trial vs Salofalk enema. The SAF-3 study group. Ital J Gastroenterol Hepatol,1999, 31(8):677.
[4]贺国斌,欧阳钦.溃疡性结肠炎的血栓前状态与肝素治疗.国外医学·内科学分册,2000,(6)：21.
[5]Hatoum OA, Spinelli KS, Abu-Hajir M, et al. Mesenteric venous thrombosis in inflammatory bowel disease. J Clin Gastroenterol,2005,39(1):27.
[6]Greenberg CS, Dcvinc DV, Mc Grac KM, et al. Measurement of plasma fibrin D-diner levels with the use of a monoclonal antibody coupled latex beed. Am J Clin Ppathol,1987,87(1):94-99.
[7]Bjerregaard LT, Nederby NT, Fredholm L, et al. Hyperhomocysteinaemia. Coagulation pathway activation and thrombophilia in patients with inflammtory bowel disease. Scand J Gastroenterol,2002,37(1):62.
[8]Collins CE, Rampton DS. Platelet dysfunction:a new dimension in inflammatory bowel disease. Gut,1995,36(1):5.
[9]Collins CE, Rampton DS. Platelets in inflammatory bowel disease pathogenetic role and therapeutic implications. Aliment Pharmacol Ther,1997,11(2):237.
[10]陈隆典,杨英.炎症性肠病疾病活动程度与血小板参数的相关性分析.中华消化杂志,2006,26(11)：771.
[11]Kapsoritakis AN, Koukourakis MI, Sfiridaki A, et al. Mean platelet volume: a useful marker of inflammatory bowel disease activity. Am J Gastroenterol,2001, 96(3):776.
[1]Morris GP, Beck PL, Herridge MS, et al. Hapte-induced model of chronic inflammation and ulceration in the rat colon. Gastroenterology,1989,96(3):795-803.
[2]锁堂,吴焕,施达仁. 白介素与溃疡性结肠炎. 世界华人消化杂志,2006,14(4)：405.
[3]Nielsen OH, Koppen T, Rudiger. Involvement of interleukirr-4 and 10 in inflammatory bowel disease. Dig Dis Sei,1996,41:1786.
[4]崔海宏,陈村龙,杨玉捷,等.溃疡性结肠炎患者肠黏膜Thl/Th2类细胞因子-mRNA的表达.世界华人消化杂志,2003,11：1524-1527.
[5]Melgar S, YeungM M, BasA, et al. Over-expression of inter-leukin 10 in mucosal T cells of patientswith active ulcerative colitis. Clin Exp Immunol,2003,134: 127-137.
[6]Dreyer C, Krey G, Keller H, et al. Control of the peroxisomal beta-oxidation pathway by a novel family of nuclear hormone receptors. Cell,1992,68(5):879-887.
[7]Meithaeghe A, Fajas L, Gouilleux F, et al. A functional polymorphism in a STA5B site of human PPAR gamma3gene promoter affects height and lipid metabolism in a French population. Arterropscler Thromb Vasc Biol,2003,23:L284.
[8]Malamut G, Cabane C, Dubuquoy L, et al.No evidence for an involvement of the p38 and JNK mitogen-activated protein in inflammatory bowel diseases. Dig Dis Sci.2006,51:1443-1453.
[9]Sanchez-Hidalgo M, Martin AR, Villegas I, et al. Rosiglitazone, a PPAR gamma ligand, modulates signal transduction pathways during the development of acute TNBS-induced colitis in rats. Eur J Pharmacol,2007,562(3):247-258.
[10]Desreumaux P, Ernst 0, Geboes K, et al. Inflammatory alterations in mesenteric adipose tissue in Crohn's disease. Gastroenterology,1999,117(1):73-81.
[11]Dubuquoy L, Jansson EA, Deeb S, et al. Impaired expression of peroxisome proliferator-activated receptor gamma in ulcerative colitis. Gastroenterology, 2003,124(5):1265-1276.
[12]Desreumaux P, Dubuquoy L, Nutten S, et al. Attenuation of colon inflammation through activators of the retinoid X receptor(RXR)/peroxisome proliferator-activated receptor gamma(PPARgamma) heterodimer:A basis for new therapeutic strategies.J Exp Med,2001,193(7):827-838.
[13]梁红亮,欧阳钦.过氧化物酶体增生物激活受体-γ在小鼠噁唑酮结肠炎中作用的研究.胃肠病学,2007,12(2)：101-105.
[14]Sen R, Baltimore D. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell,1986,46(5):705-716.
[15]Chen YG, Zhang C, Chiang SK, et al. Increased nuclear factor-kappa B p65 immunoreactivity following retinal ischemia and reperfusion injury in mice. J Neurosci Res,2003,72(1):125-131.
[16]Zaninoni A, Imperiali FG, Pasquini C, et al. Cytokine modulation of nuclear factor2kappa B activity in B2chronic lymphocytic leukemia. Exp Hematol,2003,31 (3):185-190.
[17]Chen YM, Tu CJ, Hung KY, et al. Inhibition by pentoxifylline of TNF-alpha stimulated f ractalkine production in vascular smoot h muscle cells:evidence for mediation by NF2kappa B downregulation. Br J Pharmacol,2003,138 (5):950-958.
[18]Kis A, Yellon DM, Baxter GF. Role of nuclear factor-kappa B activation in acute ischaemia reperfusion injury in myocardium. Br J Pharmacol,2003,138 (5):894-900.
[19]Calder VL, Bondeson J, Brennan FM, et al. Antigen-specific T2 cell down regulation by human dendritic cells following blockade of NF-kappaB. Scand J Immunol,2003,57(3):261-270.
[20]Abreu, Maria T. Immunologic regulation of toll-like receptors in gut epithelium. Gastroenterol,2003,19(6):559-564.
[21]Ashwood P, Harvey R, Ver jee T, et al. Funtional interactions between mucosal IL-1, IL-ra and TGF-bete 1 in ulcerative colitis. Inflamm Res,2004,53(2):53-59.
[22]Sawa Y, Oshitani N, Adachi K, et al. Comprehensive analysis of intestinal cytokine messenger RNA profile by real-time quantitative polymerase chain reaction in patient s with inflammatory bowel disease. Int J Mol Med,2003,11 (2):175-179.
[23]甘华田,欧阳钦,贾道全,等.溃疡性结肠炎患者核因子-κB活化与细胞因子基因表达.中华内科杂志,2002,41：252-255.
[24]Hashimoto C, Hudson KL, Anderson KV. The Toll Gene of Drosophila, required for dorsal-ventral embryonic polarity, appears to encode a transmembrane protein. Cell,1988,52(2):269-279.
[25]Medzhitov R, Preston-Hurlburt P, Janeway CA, et al. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature, 1997,388 (6640):394-397.
[26]Netea MG, Van derMeer JW, Kullberg BJ. Recognition of pathogenic microorganisms by Toll-like receptors. Drugs Today (Bare),2006,42 (supp lA): 99-105.
[27]Canto E, Ricart E, MonfortD, et al. TNF alphap roduction to TLR2 ligands in active IBD patients. Clin Immunol,2006,119(2):156-165.
[28]Cario E, Podolsky DK. Differential alteration in intestinal epithelial cell expression of Toll-like receptor 3 (TLR3) and TLR4 in inflammatory bowel disasel Infect Immun,2000,68(12):7010-7017.
[29]Toiyama Y, Araki T, Yoshiyama S, et al. The expression patterns of Toll-like receptors in the ileal pouch mucosa of postoperative ulcerative colitis patientsl Surg Today,2006,36(3):287-290.
[30]Douglas R, McDonald, Riny Janssen, Raif Geha. Lessons learned from molecular defects in nuclear factor κ B dependent signaling. Microbes and infection,2006,8(4):1151-1156.
[31]Hanauer, Stephen B MD. Inflammatory bowel disease: epidemiology, pathogenesis, and therapeutic opportunities. Inflammatory Bowl Disease,2006,12 (supplement 1):S3-S9.
[32]Abreu MT, Vora P, Fature E, et al. Decreased expression of Toll-like receptor-4 and MD-2correlates with intestinal epithelial cell protection against dysregulated proinflammatory gene expression in response to bacterial lipopoly saccharide. J Immunol,2001,167(3):1609-1617.
[33]Paul N, Moynag H. TLR signalling and activation of IRFs:revisiting old friends from the NF-κB pathway. Trends in immunology,2005,26(9):469-476.
[34]Liu Y, Wang Y, Yamankuchi M, et al. Upregulation of Toll-like receptor 2 gene expression in macrophage response to peptidoglycan and high concentration of lipopolysaccharide is involved in NF-κB activation. Infection Immun,2001,69 (5):2788-2796.
[35]刘一品,李延青.核因子-κB的表达在溃疡性结肠炎发病机制中的意义.胃肠病 学,2006,11(2)：103-106.
1、分析了溃疡性结肠炎湿热内蕴和血瘀之间的关系,并以现代医学高凝状态的指标验证血瘀证的病理状态,求证灌肠方的作用机理。
2、动物实验从调整细胞因子间的平衡状态、抑制自身免疫反应方面,从蛋白、基因水平多角度的分析灌肠方的作用机理,开阔了视野,拓展了中医药治疗溃疡性结肠炎的思路。
3、溃疡性结肠炎模型制作中,我们采用了双猪尾管从肛门插入,根据管上的刻度,易于掌握插入的长度；此管为软管,易于操作,并且对大鼠结肠粘膜的机械损伤小,更符合溃结的病理生理模型。