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电针通过调节肥大细胞/类胰蛋白酶/蛋白酶激活受体-2/肌球蛋白轻链激酶通路保护腹泻型肠易激综合征大鼠的肠黏膜屏障。

Electroacupuncture protects the intestinal mucosal barrier in diarrhea-predominant Irritable Bowel Syndrome rats by regulating the MCs/Tryptase/PAR-2/MLCK pathway.

作者信息

Han Xiao-Yu, Song Xiao-Ge, Ma Wen-Li, Fang Ming, Zhu Jing-Wei, Ruan Jing-Ru, Li Kui-Wu, Zou Ling, Liao Lu-Min, Li Xiao-Min, Wang Zi-Ye, Fang Yu-Cheng, Chu Hao-Ran

机构信息

Anhui University of Chinese Medicine Hefei 230031, Anhui, China.

Institute of Acupuncture and Meridians, Anhui University of Traditional Chinese Medicine Hefei 230038, Anhui, China.

出版信息

Am J Transl Res. 2024 Mar 15;16(3):781-793. doi: 10.62347/VZJL1218. eCollection 2024.

Abstract

OBJECTIVE

The pathogenesis of diarrhea-predominant irritable bowel syndrome (IBS-D) is related to damage to the intestinal mucosal barrier function. Based on the Mast cell (MC)/Tryptase/Protease-activated receptor-2 (PAR-2)/Myosin light chain kinase (MLCK) pathway, this study explored the effect of electroacupuncture (EA) on IBS-D rats and its possible mechanism of protecting the intestinal mucosal barrier.

METHODS

The IBS-D rat model was established by mother-offspring separation, acetic acid enema, and chronic restraint stress. The efficacy of EA on IBS-D rats was evaluated by observing the rate of loose stool (LSP) and the minimum volume threshold of abdominal withdrawal reflex (AWR) in rats. Mast cells and the ultrastructure of intestinal mucosa were observed by H&E staining, toluidine blue staining, and transmission electron microscopy. The expression levels of Tryptase, PAR-2, MLCK, zonula occludens-1 (ZO-1), and Occludin in rats were detected by ELISA, qRT-PCR, and western blot.

RESULTS

After 7 days of intervention, compared to the IBS-D group, the loose stool rates of rats in IBS-D + EA group and IBS-D + ketotifen group were decreased ( < 0.01), the minimum volume thresholds of AWR were improved ( < 0.01), the inflammation of colon tissue decreased, the number of MCs were decreased ( < 0.01), the expression of Tryptase, PAR-2, and MLCK were lowered ( < 0.01, < 0.05), and the expression of ZO-1 and Occludin were enhanced ( < 0.01, < 0.05). Compared to the EA group, there was no significant difference in each index between the ketotifen groups ( > 0.05).

CONCLUSION

EA has a good therapeutic effect on IBS-D rats. Regulating the MCs/Tryptase/PAR-2/MLCK pathway may be a mechanism to protect the intestinal mucosal barrier.

摘要

目的

腹泻型肠易激综合征(IBS-D)的发病机制与肠黏膜屏障功能受损有关。基于肥大细胞(MC)/组织蛋白酶/蛋白酶激活受体-2(PAR-2)/肌球蛋白轻链激酶(MLCK)通路,本研究探讨电针(EA)对IBS-D大鼠的影响及其保护肠黏膜屏障的可能机制。

方法

通过母子分离、醋酸灌肠和慢性束缚应激建立IBS-D大鼠模型。通过观察大鼠的稀便率(LSP)和腹部退缩反射(AWR)的最小容量阈值来评估EA对IBS-D大鼠的疗效。采用苏木精-伊红染色、甲苯胺蓝染色和透射电子显微镜观察肥大细胞和肠黏膜超微结构。采用酶联免疫吸附测定(ELISA)、实时荧光定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测大鼠组织蛋白酶、PAR-2、MLCK、紧密连接蛋白-1(ZO-1)和闭合蛋白的表达水平。

结果

干预7天后,与IBS-D组相比,IBS-D + EA组和IBS-D + 酮替芬组大鼠的稀便率降低(<0.01),AWR的最小容量阈值提高(<0.01),结肠组织炎症减轻,MC数量减少(<0.01),组织蛋白酶、PAR-2和MLCK的表达降低(<0.01,<0.05),ZO-1和闭合蛋白的表达增强(<0.01,<0.05)。与EA组相比,酮替芬组各指标差异无统计学意义(>0.05)。

结论

EA对IBS-D大鼠有良好的治疗作用。调节MCs/组织蛋白酶/PAR-2/MLCK通路可能是保护肠黏膜屏障的机制之一。

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