肖瑶 刘百祥 封迎帅 余阗 吴海金 袁利群

〔摘要〕 目的 觀察加味椒艾丸贴敷神阙穴对轮状病毒腹泻小鼠NOD1/NF-κB信号通路的影响，探讨其干预轮状病毒感染性腹泻的机制。方法 将40只SPF级ICR乳鼠中的10只分为正常对照组，其余30只采用猴轮状病毒SA11株悬液灌胃造模2 d，造模成功后随机分为模型组、丁桂贴组、加味椒艾丸组，每组10只。药物干预敷贴连续5 d，末次给药并禁食12 h后取材。收集各组小鼠粪便进行粪便评分;ELISA检测各组血清TNF-α、IL-1β、Caspase-1含量;Western blot检测各组小鼠小肠组织p65、NF-κB蛋白表达;HE染色观察各组小鼠肠组织病理形态。结果 （1）与正常对照组比较，模型组小鼠粪便评分明显升高（P<0.01）;与模型组比较，丁桂贴组和加味椒艾丸组粪便评分明显降低（P<0.05）。（2）与正常对照组比较，模型组血清中TNF-α、Caspase-1含量明显升高（P<0.01）;与模型组比较，丁桂贴组及加味椒艾丸组TNF-α、Caspase-1含量明显下降（P<0.05，P<0.01）;与加味椒艾丸组比较，丁桂贴组Caspase-1含量明显降低（P<0.01）。（3）与正常对照组比较，模型组p65、NF-κB蛋白表达升高（P<0.01）;与模型组比较，丁桂贴组、加味椒艾丸组p65、NF-κB蛋白表达降低（P<0.05，P<0.01）。（4）正常对照组小鼠回肠和结肠黏膜上皮未见异常;模型组小鼠回肠和结肠黏膜上皮变性坏死，部分黏膜上皮缺失，且伴有不同程度的黏膜层血管扩张;丁桂贴组和加味椒艾丸组小鼠回肠和结肠黏膜上皮部分缺失，黏膜层血管少量扩张。结论 加味椒艾丸贴敷神阙穴能改善轮状病毒腹泻症状，可能通过降低血清中TNF-α、Caspase-1含量及小肠组织中p65、NF-κB蛋白表达来调控NOD1/NF-κB信号通路。

〔关键词〕 轮状病毒腹泻;加味椒艾丸;神阙穴;敷贴;NOD1/NF-κB信号通路;粪便评分

〔中图分类号〕R256.34      〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.07.012

〔Abstract〕 Objective To observe the effect of Jiawei Jiaoai Pills applied to Shenque （RN8） acupoint on NOD1/NF-κB signalling pathway of rotavirus diarrhea rats， and to explore the mechanism in treating rotavirus diarrhea. Methods The 10 of 40 SPF neonatal rat randomly selected as the control group， the remaining 30 rats by gastric perfusion of suspension of simian rotavirus SA11 strain for two days to establishment of rotavirus diarrhea rat model， which rats were randomly divided into the model group， Dinggui application group and Jiawei Jiaoai Pills group， with 10 rats in each group. The drug intervention application continued for 5 days. The samples were taken from the 12 hours after the last acupoint application. The feces of mice in each group were collected for diarrhea score; the content of serum TNF-α， IL-1β， Caspase-1 was detected by ELISA; the expression levels of p65， NF-κB were analyzed by Western blot; HE staining was used to observe the pathological morphology of intestinal tissues in each group. Results （1） Compared with the control group， faeces score of model group was significantly increased （P<0.01）; compared with the model group， faeces score was obvious decreased in the Dinggui application group and Jiawei Jiaoai Pills group （P<0.05）. （2） Compared with the contorl group， the levels of serum TNF-α， Caspase-1 were significantly reduced in model group （P<0.01）; compared with the model group， the levels of serum TNF-α， Caspase-1 were obvious decline in the Dinggui application group and Jiawei Jiaoai Pills group （P<0.05， P<0.01）; compared with Jiawei Jiaoai Pills group， the level of Caspase-1 was declined in the Dinggui application group （P<0.01）. （3） Compared with the control group， the expression levels of p65， NF-κB were higher in model group （P<0.01）; compared with the model group， the expression levels of p65， NF-κB were declined in the Dinggui application group and Jiawei Jiaoai Pills group （P<0.05， P<0.01）. （4） There was no abnormality in the ileum and colon mucosal epithelium in the control group， while the ileum and colon mucosal epithelium in the model group was degenerated and necrotic， with partial loss of mucosal epithelium and accompanied by different degrees of vascular dilatation in the mucosal layer. Mucosal epithelium of ileum and colon in Dinggui application group and Jiawei Jiaoai Pills group were partially absent， and blood vessels of mucosal layer were slightly dilated. Conclusion Jiawei Jiaoai Pills applied to Shenque （RN8） acupoint can improve the diarrhea symptoms of rotavirus diarrhea of rats， and regulate NOD1/NF-κB signaling pathway by down regulating the protein expression of serum TNF-α， Caspase-1 and intestinal tissue p65， NF-κB.

〔Keywords〕 rotavirus diarrhea; Jiawei Jiaoai Pills; Shenque （RN8） acupoint; NOD1/NF-κB signaling pathway; faeces score

轮状病毒（rotavirus， RV）是全球婴幼儿腹泻最常见的病原体，是儿童腹泻的主要原因，尤其是3岁以下儿童发病率较高[1-2]。RV感染的急性腹泻婴幼儿会伴有明显肠道菌群失衡、血清炎症因子水平升高现象，其发病主要原因是自身免疫功能降低，病原微生物入侵，导致炎症感染，RV感染性肠炎在一定程度上会导致呼吸、心肌及神经系统损害[3-4]。目前，对于RV腹泻并无特效药物，近年来，研究[5-6]表明，中医药在治疗腹泻方面具有一定的临床疗效。治疗RV腹泻的早期临床研究[7-8]表明，加味椒艾丸穴位贴敷涌泉穴或神阙穴治疗婴幼儿RV腹泻较贴脐疗效更优，能缩短病程，并且在治疗婴幼儿急性腹泻的临床疗效优于单纯西医治疗。但加味……