艾灸神阙穴对溃疡性结肠炎小鼠血清促炎性因子及肠黏膜紧密连接蛋白的影响

Effect of Moxibustion at Shenque(RN8)on Serum Proinflammatory Factors and Colon Mucosa Tight Junction Protein in Mice with Ulcerative Colitis

目的观察艾灸神阙穴对DSS诱导的溃疡性结肠炎(UC)小鼠的疗效,探讨艾灸神阙穴对UC小鼠炎性反应及肠黏膜屏障的影响。方法以32只雄性ICR小鼠为研究对象,随机选取8只作为空白对照组,其余24只通过自由饮用3.5%DSS溶液8 d制成UC模型,造模成功后再随机分为模型组、艾灸组和阳性药物组,每组8只。持续干预8 d:艾灸组小鼠每日抓取放入固定器固定,艾灸神阙穴20 min,每日1次;空白对照组和模型组每日进行同艾灸组同样的抓取固定,但不进行其他操作,每日1次;阳性药物组小鼠按0.4 g/kg体质量灌服美沙拉嗪溶液,每日1次。干预结束后取小鼠血清用ELISA法检测促炎性因子白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)表达水平,取小鼠结肠组织用Western blotting法检测紧密连接蛋白1(ZO-1)、Occludin蛋白表达水平,采用HE染色法观察结肠组织病理切片,通过疾病活动指数评分(DAI)观测UC小鼠的恢复情况。结果与空白对照组相比,UC模型小鼠体质量减轻,大便稀软并带有脓血便,模型组小鼠DAI评分显著升高(P<0.05),病理组织切片显示结肠黏膜组织严重损伤,有明显炎性反应,促炎性因子IL-6、TNF-α含量明显升高(P<0.05),结肠组织中ZO-1、Occludin蛋白表达均显著降低(P<0.01)。与模型组相比,艾灸组与阳性药物组小鼠结肠组织病理损伤有显著改善,炎性反应下调,DAI评分显著降低(P<0.05),血清中IL-6、TNF-α含量显著降低(P<0.05),结肠组织中ZO-1、Occludin蛋白表达均显著升高(P<0.01)。结论艾灸神阙穴能有效缓解UC症状,升高结肠组织中紧密连接蛋白ZO-1、Occludin的表达,改善结肠组织损伤,降低UC小鼠血清炎性因子IL-6和TNF-α含量,对UC小鼠有治疗作用;提示艾灸对UC的起效机制可能与肠上皮黏膜屏障有关。

Objective:To observe the curative effect of moxibustion at Shenque(RN8)on DSS-induced UC mice,and to explore the effect of moxibustion at Shenque(RN8)on inflammatory response and intestinal mucosal barrier in ulcerative colitis mice.Methods:32 male ICR mice as the research object,8 randomly selected as blank control group,the remaining 24 were made UC model by freely drinking 3.5%DSS solution for 8 days,and then randomly divided into 3 groups:model group,moxibustion group and the positive drug group,there are 8 rats in each group,and each group intervenes in the following manner for 8 days:the moxibustion group mice are grabbed and fixed in a fixture every day,and moxibustion at Shenque(RN8)for 20 min/d,once a day;The blank control group and the model group were treated with the same grasping and fixing as the moxibustion group daily,but no other operations were performed,once a day;the mice in the positive drug group were given mesalazine solution at a dose of 0.4 g/kg body weight,once a day..After the intervention,the serum of mice was taken to detect IL-6 and TNF-αby ELSA kit.The expression of ZO-1 and Occludin protein was detected by Western blotting.The pathological sections of colon tissue were observed by HE staining.Observe the recovery of UC mice by disease activity index score(DAI).Result:Compared with the blank control group,the UC model mice lost weight,had soft stools with pus and bloody stools,and the model group had significantly higher DAI scores(P<0.05).Inflammatory response,proinflammatory factors IL-6 and TNF-αlevels were significantly increased(P<0.05),and the expression of ZO-1 and Occludin protein in colon tissue were significantly reduced(P<0.01).Compared with the model group,the pathological damage of the colon tissue in the moxibustion group and the positive drug group was significantly improved,the inflammatory response was reduced,the DAI score was significantly reduced(P<0.05),and the levels of IL-6 and TNF-αin serum were significantly reduced(P<0.05),the expression of ZO-1 and Occludin protein in colon tissue were significantly increased(P<0.01).Conclusion:Moxibustion at Shenque(RN8)can effectively relieve UC symptoms,increase the expression of tight junction proteins ZO-1 and Occludin in colon tissue,improve colon tissue damage,and reduce the content of serum inflammatory factors IL-6 and TNF-αin UC mice.Moxibustion has a therapeutic effect on UC mice.It is suggested that the effect mechanism of moxibustion on UC may be related to intestinal epithelial mucosal barrier.