艾灸对克罗恩病大鼠结肠NF-κB p65及TNF-α、IL-1β调节作用的研究

Study on the Regulating Effect of Moxibustion on NF-κB p65,TNF-α,and IL-1β in Colons of CD Rats

目的:通过观察隔药灸对克罗恩病大鼠结肠NF-κB p65及炎症因子TNF-α、IL-1β表达的影响,探讨艾灸治疗克罗恩病的效应机制。方法:清洁级雄性SD大鼠随机分为4组,分别为正常组、模型组、隔药灸组和西药组,采用三硝基苯磺酸(TN-BS)合乙醇溶液灌肠制备克罗恩病大鼠模型,模型制备成功后,隔药灸组取天枢穴(双)、气海穴进行隔药灸治疗,西药组采用美沙拉秦灌胃治疗。治疗结束后,采用HE染色,光镜下观察各组大鼠结肠组织病理学变化,应用免疫组织化学技术观察各组大鼠结肠NF-κB p65与TNF-α、IL-1β的表达,并进行图像分析。结果:与正常组比较,模型组大鼠结肠组织呈现裂隙状溃疡及炎症表现,部分大鼠结肠在黏膜下层可见肉芽肿形成,结肠NF-κB p65、TNF-α、IL-1β的表达均显著增高(均P<0.01);与模型组比较,隔药灸组、西药组大鼠结肠形态结构改善,肠道炎症减轻,结肠NF-κB p65与TNF-α、IL-1β的表达均显著降低(均P<0.01);相关性分析结果显示,TNF-α、IL-1β的表达与NF-κB p65的表达均呈正相关性。结论:隔药灸能下调克罗恩病大鼠结肠NF-κB p65、TNF-α、IL-1β的表达;艾灸(隔药灸)可能通过抑制克罗恩病大鼠结肠NF-κB p65的表达,进而减少其下游炎症因子TNF-α、IL-1β的表达,从而减轻肠道炎症,改善结肠组织形态结构,发挥治疗作用。

Objective：To explore the action mechanism of moxibustion in treating Crohn＇s disease（CD）,by observing the effect of herb-partitioned moxibustion on the expressions of NF-κB p65,TNF-α,and IL-1β.Methods：Male Sprague Dawley（SD） rats of clean conventional grade were randomized into four groups,i.e.a normal control（NC） group,a model control（MC） group,a herb-partitioned moxibustion（HPM） group,and a Western medication（WM） group.The CD rat models were developed by using the mixture of 2,4,6 trinitrobenzene sulfonic acid（TNBS） and alcohol via enema.When the models were successfully developed,the HPM group received HPM intervention at bilateral Tianshu（ST 25） and Qihai（CV 6）,and the WM group received Mesalazine via intragastric administration.At the end of treatment,the histopathological variations of rats＇ colons were observed by adopting HE staining and light microscope,and the expressions of NF-κB p65,TNF-α,and IL-1β in rats＇ colons were determined by using immunohistochemical technique,and the images were all analyzed.Results：Compared to the NC group,the MC group had its rats＇ colons present with fissured ulcers and inflammation,and granulomas in submucosa of some colons,and its expressions of NF-κB p65,TNF-α,and IL-1β increased significantly（P 0.01） ;compared to the MC group,the HPM group and WM group both had rats＇ colons present with improved structures and reduced intestinal inflammation,and their expressions of NF-κB p65,TNF-α,and IL-1β dropped significantly（P 0.01） ;the relevance analyses showed that the expressions of TNF-α and IL-1β both had a positive correlation with the expression of NF-κB p65.Conclusion：HPM can down-regulate the expressions of NF-κB p65,TNF-α,and IL-1β in CD rats＇ colons;it＇s possible that moxibustion（HPM） produces its therapeutic efficacy by inhibiting the expression of NF-κB p65 in rat＇s colon,and subsequently decreasing the expressions of its downstream inflammatory factors,i.e.TNF-α and IL-1β,by which,the intestinal inflammation is relieved and the morphology of colon is improved.