艾灸对溃疡性结肠炎大鼠结肠Ubiquitin及NLRP3蛋白的调节作用

目的:观察艾灸对溃疡性结肠炎(UC)大鼠结肠炎症和泛素(ubiquitin)、核苷酸结合寡聚化结构域(Nod)样受体蛋白3(NLRP3)蛋白表达的影响,探讨艾灸治疗UC的抗炎机制。方法:清洁级雄性Sprague-Dawley(SD)大鼠随机分为正常组、模型组、艾条灸组和西药组。采用自由饮用35g/L葡聚糖硫酸钠(DSS)溶液制备大鼠UC模型。艾条灸组取双侧天枢穴,给予温和灸治疗;西药组给予美沙拉嗪溶液灌胃治疗。正常组和模型组不进行任何治疗,只做与艾条灸组相同的抓取和固定。治疗结束后,采用苏木素-伊红(HE)染色,光镜下观察结肠组织病理损伤并评分;采用免疫荧光法测定结肠ubiquitin蛋白的表达;运用兔疫组织化学法检测结肠白细胞介素(IL)-1β和NLRP3蛋白的表达。结果:与正常组比较,模型组大鼠结肠组织损伤严重,病理评分明显升高(P<0.01),结肠ubiquitin、NLRP3及IL-1β蛋白表达均显著增强(均P<0.01)。与模型组比较,艾条灸组及西药组大鼠结肠损伤有所修复,炎症反应减轻,病理评分均降低(均P<0.01);ubiquitin、NLRP3及IL-1β蛋白的表达减少(均P<0.01)。相关分析显示,结肠组织ubiquitin蛋白与结肠病理评分、NLRP3蛋白的表达之间存在相关性(r=0.677,P<0.01;r=0.536,P<0.05).结论:艾灸能下调UC大鼠结肠ubiquitin、NLRP3和IL-IP蛋白的表达,该作用可能是其促进结肠炎性损伤修复、发挥抗炎作用的机制之一。

Regulatory effects of moxibustion on ubiquitin and NLRP3 proteins in colon of ulcerative colitis rats

Objective:To observe the effects of moxibustion on colonic inflammation,and the expressions of ubiquitin and nucleotide-binding oligomerization domain(Nod)-Iike receptor protein 3(NLRP3)proteins in rats with ulcerative colitis(UC),and to explore the anti-inflammatory mechanism of moxibustion in the UC treatment.Methods:Clean grade male Sprague-Dawley(SD)rats were randomly divided into a normal group(NG),a model group(MG),a moxa-stick moxibustion group(MSMG)and a Western medicine group(WMG).UC model was prepared by freely drinking 35 g/L d ext ran sulfate sodium(DSS)solution.Bilateral Tianshu(ST 25)were selected for mild moxibustion treatment in the MSMG;mesalazine solution was intragastrically administrated in the WMG.Rats in the NG and MG were only grasped and fixed as in the MSMG without any treatment.After treatment,hematoxylin-eosin(HE)staining was performed to observe and score the colonic pathological damage under light microscope;immunofluorescence method was used to determine the expression of colonic ubiquitin protein;immunohistochemical method was used to detect the expressions of colonic interleukin(IL)-1β and NLRP3 proteins.Results:The colon tissue was severely injured,and the pathological score was significantly increased in the MG than in the NG(P<0.01),and the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon were significantly increased(all P<0.01).Compared with the MG,the colonic damage was repaired,the inflammation and pathological scores were reduced,and the ubiquitin,NLRP3 and IL-1β protein expressions were decreased in the MSMG and WMG(all P<0.01).Correlation analysis revealed that the ubiquitin protein expression was correlated with the colonic pathological score and the NLRP3 protein expression(r=0.677,P<0.01;r=0.536,P<0.05).Conclusion:Moxibustion can down-regulate the protein expressions of ubiquitin,NLRP3 and IL-1β in the colon of UC rats,which may be one of the mechanisms to promote the repair of colonic inflammatory lesions and exert anti-inflammatory effects.