埋线联合艾灸对溃疡性结肠炎活动期大鼠结肠黏膜Notch信号通路的调控作用

Effect of acupoint catgut embedding combined with moxibustion on Notch signaling pathway in colonic mucosa of rats with active ulcerative colitis

目的:观察穴位埋线联合艾灸对溃疡性结肠炎(UC)大鼠结肠黏膜组织Notch受体1及靶基因Hes 1、Math 1表达的影响,从Notch信号通路角度探讨其治疗UC的机制。方法:SD大鼠随机分为正常组、模型组、埋线+艾灸组、埋线组、艾灸组,每组6只。采用5%三硝基苯磺酸+50%乙醇混合溶液灌肠法复制UC模型。模型成功次日开始治疗,穴位取"上巨虚""天枢""大肠俞"。艾灸组艾条温和灸10 min, 1次/d,共14次。埋线组用简易埋线针进行穴位埋线,1次/周,共2次。埋线+艾灸组于艾灸后6 h埋线。HE染色法观察结肠黏膜组织形态变化;q-PCR法和Western blot法检测结肠黏膜组织Notch 1、Hes 1和Math 1 mRNA及蛋白表达。结果:正常组大鼠结肠上皮完整、结构清晰,杯状细胞丰富,腺管结构正常。模型组大鼠结肠黏膜结构模糊,上皮大量破损、脱落,杯状细胞缺失,腺管破坏,大量炎性细胞浸润,各治疗组较模型组结肠损伤有不同程度改善,埋线+艾灸组最为明显。与正常组比较,模型组结肠黏膜组织Notch 1、Hes 1 mRNA及蛋白表达水平升高(P<0.01),Math 1 mRNA及蛋白表达水平降低(P<0.01);与模型组比较,各治疗组结肠黏膜组织Notch 1、Hes 1 mRNA及蛋白表达水平降低(P<0.01,P<0.05),埋线+艾灸组、埋线组Math 1 mRNA表达水平升高(P<0.01),各治疗组结肠黏膜组织Math 1蛋白表达水平升高(P<0.01)。埋线+艾灸组结肠黏膜组织Notch 1、Hes 1 mRNA及蛋白表达水平低于埋线组、艾灸组(P<0.01,P<0.05),Math 1 mRNA表达水平高于艾灸组(P<0.01),Math 1蛋白表达水平高于埋线组、艾灸组(P<0.01)。结论:穴位埋线+艾灸对UC大鼠结肠损伤修复具有促进作用,其可能通过降低结肠黏膜组织Notch 1、Hes 1表达水平,上调Math 1表达水平,抑制Notch信号通路的过度活化,从而调节结肠上皮分泌细胞系和吸收细胞系间分化的平衡而起效。

Objective To observe the effect of acupoint catgut embedding(ACE) combined with moxibustion on the expressions of Notch receptor 1 and target genes Hes 1 and Math 1 in colonic mucosa of ulcerative colitis(UC) rats, so as to reveal its mechanism of treating UC from the perspective of Notch signaling pathway. Methods Thirty SD rats were randomized into normal, model, ACE combined with moxibustion(combination), ACE, and moxibustion groups(n=6 in each group). The UC model was established by intrarectal administration of 5% trinitro-benzene-sulfonic acid and 50% ethyl alcohol. Moxibustion was applied to bilateral "Shangjuxu"(ST37), "Tianshu"(ST25) and "Dachangshu"(BL25) for 10 min, once daily for 14 days, and ACE applied to the same 3 acupoints, once a week for two weeks. After the treatment, colonic mucosal pathological changes were observed after hematoxylin eosin(HE) stain. Protein and mRNA expressions of Notch 1,Hes 1 and Math 1 were detected by RT-PCR and Western blot, respectively. Results H.E. staining showed severe defect of the colonic mucosal epithelium with infiltration of a large number of inflammatory cells in the model group, which was milder in moxibustion, ACE and moxibustion + ACE groups. After modeling, the protein and mRNA expressions of Notch 1 and Hes 1 in the colonic mucosa were obviously increased(P<0.01), while the protein and mRNA expressions of Math 1 were significantly decreased(P<0.01) in the model group relevant to the normal group. Following the intervention, the protein and mRNA expression levels of Notch 1 and Hes 1 in the colonic mucosa were significantly lower(P<0.05,P<0.01) in moxibustion, ACE and combination groups, the mRNA expression level of Math 1 was significantly increased in both ACE and combination groups(P<0.01), and the protein expression of Math 1 was significantly higher in the three treatment groups relevant to the model group(P<0.01). The above indexes of moxibustion + ACE were considerably superior to those of simple ACE and simple moxibustion(P<0.05, P<0.01). Conclusion Acupoint catgut embedding combined with moxibustion can repair the injured colonic mucosa of UC rats, which may be related with its effects in suppressing the expression of Notch 1 and Hes 1, up-regulating the expression of Math 1 in the colonic mucosa, thereby regulating the balance of differentiation between colonic epithelial secretory and absorption cell lines.