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Study on the regulatory effect of herbal cake-partitioned moxibustion on colonic CD206, AMPK and TSC2 in rats with Crohn disease

隔药灸对克罗恩病大鼠结肠CD206, AMPK和TSC2的调节作用研究

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Abstract

Objective

To explore the mechanism of herbal cake-partitioned moxibustion in Crohn disease (CD) treatment by observing the effect of herbal cake-partitioned moxibustion on protein expressions of colonic M2 macrophage marker CD206, AMP-activated protein kinase (AMPK) and tuberous sclerosis complex (TSC) 2.

Methods

Twenty-six specific pathogen free male rats were randomly divided into a normal group, a model group and a herbal cake-partitioned moxibustion group. The CD model was prepared by enema with the mixture of 5% (W/V) 2,4,6-trinitrobenzene sulfonic acid (TNBS) and 50% ethanol at 2:1 (volume ratio). After the model was successfully prepared, rats in the herbal cake-partitioned moxibustion group received herbal cake-partitioned moxibustion at Qihai (CV 6) and bilateral Tianshu (ST 25). Hematoxylin-eosin (HE) staining was used to observe the histopathological changes of rat colon; immunohistochemical technique was used to detect the expression of colonic CD206 protein; Western blot, immunofluorescence, and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) technologies were used to detect the protein and mRNA expressions of colonic AMPK and TSC2.

Results

Compared with the normal group, rats in the model group showed damaged colonic mucosa, missing of the epithelial layer, thickened submucosa, vascular proliferation, massive infiltration of monocytes and lymphocytes, and cracked ulcers that reached the muscle layer. Rats in the herbal cake-partitioned moxibustion group showed reduced intestinal inflammation and healing intestinal epithelium ulcers. Compared with the normal group, rat colonic CD206 protein expression, and the protein and mRNA expressions of colonic AMPK and TSC2 were decreased in the model group (all P<0.01); compared with the model group, rat colonic CD206 protein expression was increased (P<0.01), as well as the protein and mRNA expressions of AMPK and TSC2 in the herbal cake-partitioned moxibustion (all P<0.05).

Conclusion

Herbal cake-partitioned moxibustion can reduce intestinal inflammation in CD rats, increase colonic CD206 protein expression, and up-regulate the protein and mRNA expressions of colonic AMPK and TSC2.

摘要

目的

观察隔药灸对克罗恩病(CD)大鼠结肠M2型巨噬细胞标志物CD206, 腺苷酸活化蛋白激酶(AMPK)和结节 硬化复合物(TSC) 2蛋白表达的影响, 探讨隔药灸治疗CD的作用机制。

方法

将26只无特定病原体级雄性大鼠随机分 为正常组, 模型组和隔药灸组。采用5%(W/V) 2,4,6-三硝基苯磺酸(TNBS)与50%乙醇按2:1(体积比)混合后灌肠, 制备 CD模型。模型制备成功后, 隔药灸组取气海及双侧天枢进行隔药灸。采用苏木素-伊红(HE)染色观察大鼠结肠组织病 理学变化; 应用免疫组化技术检测结肠CD206蛋白表达; 应用免疫印迹, 免疫荧光和实时荧光定量聚合酶链反应 (RT-qPCR)技术检测结肠AMPK, TSC2蛋白和mRNA表达。

结果

与正常组比较, 模型组大鼠结肠黏膜受损, 上皮层缺失, 黏膜下层增厚, 血管增生, 单核细胞和淋巴细胞大量浸润, 出现深达肌层的裂隙状溃疡; 隔药灸组大鼠肠道炎症减轻, 肠上皮可见愈合性溃疡。与正常组比较, 模型组大鼠结肠CD206蛋白, AMPK和TSC2蛋白及mRNA表达均降低 (均P<0.01); 与模型组比较, 隔药灸组大鼠结肠CD206蛋白表达增高(P<0.01), AMPK, TSC2蛋白及mRNA表达 均升高(均P<0.05)。

结论

隔药灸能减轻CD大鼠肠道炎症, 提高结肠CD206 蛋白表达, 上调结肠AMPK, TSC2 蛋白及 mRNA表达。

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Acknowledgments

This work was supported by National Natural Science Foundation of China (国家自然科学基金项目, No. 81674073, No. 81904303); Natural Science Foundation of Shanghai (上海市自然科学基金项目, No. 20ZR1453000, No. 19ZR1451600).

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Authors and Affiliations

  1. Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Dong Xiao-qing  (董小庆), Li Xiao-ying  (李晓莹), Wang Xue-jun  (王雪君), Guo Xiao-cong  (郭潇聪), Long Jun-yi  (龙俊燚), Lu Yun-qiong  (卢云琼), Liu Li  (刘力), Caoyao Jia-ni  (曹姚佳妮), Wu Huan-gan  (吴焕淦) & Ma Xiao-peng  (马晓芃)

  2. Xi’an Traditional Chinese Medicine Hospital of Encephalopathy, Shaanxi Province, Xi’an, 710032, China

    Dong Xiao-qing  (董小庆)

  3. Shanghai Research Institute of Acupuncture and Meridian, Shanghai, 200030, China

    Zhang Dan  (张丹), Lu Yuan  (陆嫄), Wu Huan-gan  (吴焕淦), Xie Chen  (谢晨) & Ma Xiao-peng  (马晓芃)

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  1. Dong Xiao-qing  (董小庆)
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Corresponding authors

Correspondence to Xie Chen  (谢晨) or Ma Xiao-peng  (马晓芃).

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Co-first Authors

Dong Xiao-qing, M.M., resident physician; Li Xiao-ying, 2019 master degree candidate

Conflict of Interest

Authors Ma Xiao-peng and Wu Huan-gan are members of the Editorial Board of Journal of Acupuncture and Tuina Science. The paper was handled by other editors and has undergone rigorous peer review process. Authors Ma Xiao-peng and Wu Huan-gan were not involved in the journal’s review or decisions related to this manuscript.

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Xiao-qing, D., Xiao-ying, L., Xue-jun, W. et al. Study on the regulatory effect of herbal cake-partitioned moxibustion on colonic CD206, AMPK and TSC2 in rats with Crohn disease. J. Acupunct. Tuina. Sci. 19, 329–337 (2021). https://doi.org/10.1007/s11726-021-1263-7

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