隔药饼灸通过抑制RhoA/ROCK1/MLC通路改善克罗恩病相关性肠纤维化
Herb-Partitioned Moxibustion Improves Crohn's Disease-Associated Intestinal Fibrosis by Suppressing the RhoA/ROCK1/MLC Pathway.
作者信息
Zhao Min, Weng Zhijun, Huang Yan, Zheng Handan, Han Dong, Shen Jiacheng, Huang Rong, Liu Huirong, Wu Luyi
机构信息
Key Laboratory of Acupuncture and Immunological Effects, Shanghai University of Traditional Chinese Medicine, Shanghai 200030, China.
Shanghai Research Institute of Acupuncture and Meridian, Shanghai 200030, China.
出版信息
Evid Based Complement Alternat Med. 2021 Nov 17;2021:2247953. doi: 10.1155/2021/2247953. eCollection 2021.
BACKGROUND AND AIMS
Intestinal fibrosis is one of the severe and common complications of Crohn's disease (CD), but the etiology and pathogenesis remain uncertain. The study intended to examine whether the effect of herb-partitioned moxibustion on rats with CD-associated intestinal fibrosis is associated with the RhoA/ROCK1/MLC pathway.
METHODS
All experimental rats were randomly allocated into the normal control group (NC), model control group (MC), and herb-partitioned moxibustion group (HPM). Intestinal fibrosis was established in rats with CD by repeated rectal administrations of 2,4,6-trinitrobenzenesulfonic acid (TNBS). Herb-partitioned moxibustion was applied at the Qihai (CV6) and Tianshu (ST25) acupoints once daily for 10 days in the HPM group. In this study, histological changes were examined by hematoxylin and eosin (HE) staining; then, Masson's trichrome staining was used to assess the degree of fibrosis in each group. Experimental methods of immunohistochemistry, western blotting, and real-time PCR were applied to detect the levels of -SMA, collagen III, RhoA, ROCK1, and p-MLC. Moreover, the double immunofluorescent staining for the colocalization of both -SMA and ROCK1 was performed.
RESULTS
Contrasted with the normal controls, the collagen deposition and fibrosis scores were increased in colonic tissue of model rats, and HPM decreased the collagen deposition and fibrosis scores. The protein of -SMA and collagen III in the MC group exceeds that of the NC group; HPM decreased the expression of -SMA and collagen III in rats with intestinal fibrosis. Similarly, the expression of RhoA, ROCK1, and p-MLC in model rats was obviously increased compared with normal controls; the expression of RhoA, ROCK1, and p-MLC was decreased after HPM. The coexpression of -SMA and ROCK1 in rats with intestinal fibrosis was higher than normal rats.
CONCLUSION
HPM improves CD-associated intestinal fibrosis by suppressing the RhoA/ROCK1/MLC pathway.
背景与目的
肠道纤维化是克罗恩病（CD）严重且常见的并发症之一，但其病因和发病机制仍不明确。本研究旨在探讨隔药灸对CD相关性肠道纤维化大鼠的影响是否与RhoA/ROCK1/MLC通路有关。
方法
将所有实验大鼠随机分为正常对照组（NC）、模型对照组（MC）和隔药灸组（HPM）。通过反复直肠注射2,4,6-三硝基苯磺酸（TNBS）建立CD大鼠肠道纤维化模型。隔药灸组每日在气海（CV6）和天枢（ST25）穴位施灸1次，共10天。本研究采用苏木精-伊红（HE）染色观察组织学变化；然后，用Masson三色染色评估各组纤维化程度。应用免疫组化、蛋白质印迹法和实时荧光定量PCR实验方法检测α-SMA、Ⅲ型胶原、RhoA、ROCK1和p-MLC的水平。此外，进行α-SMA和ROCK1共定位的双重免疫荧光染色。
结果
与正常对照组相比，模型大鼠结肠组织中胶原沉积和纤维化评分增加，隔药灸降低了胶原沉积和纤维化评分。MC组α-SMA和Ⅲ型胶原的蛋白含量超过NC组；隔药灸降低了肠道纤维化大鼠α-SMA和Ⅲ型胶原的表达。同样，模型大鼠中RhoA、ROCK1和p-MLC的表达明显高于正常对照组；隔药灸后RhoA、ROCK1和p-MLC的表达降低。肠道纤维化大鼠中α-SMA和ROCK1的共表达高于正常大鼠。
结论
隔药灸通过抑制RhoA/ROCK1/MLC通路改善CD相关性肠道纤维化。
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