Effect of Oxymatrine Mediated by RhoA/ROCK Signaling Pathway on Expression of E-cadherin and TGF-β in Ulcerative Colitis
10.13422/j.cnki.syfjx.20190638
- VernacularTitle: 氧化苦参碱调控RhoA/ROCK信号通路介导溃疡性结肠炎E-cadherin及TGF-β的影响
- Author:
Yi-fan WANG
1
;
Heng FAN
1
Author Information
1. Wuhan Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
- Publication Type:Research Article
- Keywords:
oxymatrine;
ulcerative colitis;
RhoA/Rho-associated kinase (ROCK)signaling pathway;
E-cadherin;
transforming growth factor-β
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(6):73-80
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanism of Oxymatrine on epithelial-mesenchymal transition mediated by RhoA/Rho-associated kinase(ROCK) signaling pathway to prevent and treat ulcerative colitis(UC) and its related canceration by detecting the changes of ROCK, E-cadherin and transforming growth factor-β(TGF-β)in colon tissues of mice. Method:Totally 48 male Balb/c mice were randomly divided into normal control group, model group, low, medium and high-dose Oxymatrine groups (25,50,100 mg·kg-1)and Y-27632 group(10 mg·kg-1), with 8 mice in each group. Mice in control group received distilled water, while all the other mice were treated with 3% dextra sulfate sodium for 7 days to induce the ulcerative colitis model. Since the first day of modeling,Y-27632(10 mg·kg-1)and different doses of Oxymatrine(25, 50, 100 mg·kg-1) were intraperitoneally injectedfor 7 days, and equal volume of PBS was intraperitoneally injected in normal group and model group. Body weight loss, stool consistency and fecal blood loss were observed on a daily basis. On the 8thday, mice were put to death,colon was collected and its length was measured; the scores of disease activity index (DAI) were evaluated; part of the colons were fixed and stained with hematoxylin and eosin (HE) for a histopathological analysis; the ultrastructural changes of mucosa tissue in ulcerative colitis were observed by transmission electron microscope. The expression levels of TGF-β in tissue mucosa were tested by enzyme-linked immunosorbentassay(ELISA). The expression levels of Rho-associated kinase-1, Rho-associated kinase-2, E-cadherin and TGF-β in colon were measured by Western blot and Real-time PCR. Result:Compared with normal group, model group showed the infiltration of a large number of inflammatory cells in mucosa and submucosa, disordered gland arrangement, varying degrees of intestinal mucosal defect and even ulcer formation. Under electron microscopy, microvilli were sparse on the surface of intestinal epithelial cells, the gap between cell junctions was widened, goblet cells were reduced and organelles were swollen. The disease activity index,and the expression levels of ROCK-1 and ROCK-2 proteins in the colonic mucosa of model group were increased(P<0.01), while the colon length and the protein and mRNA contents of E-cadherin, TGF-β were decreased(P<0.01). Compared with model group, there were different degrees of alleviations in pathological manifestationsunder the light and electron microscopy in each treatment group. DAI score and colon length reduction were significantly decreased in each treatment group (P<0.01). The proteins and mRNA expression levels of colonic mucosa ROCK-1 and ROCK-2 of the treatment group were decreased(P<0.01), while the protein and mRNA expression levels of E-caherin and TGF-β were increased(P<0.01), which was statistically significant compared with model group. Compared with middle-dose Oxymatrine group, the ROCK-1 and ROCK-2 protein and mRNA levels were significantly increased in low-dose and high-dose groups(P<0.05,P<0.01), and the TGF-β and E-cadherin protein and mRNA levels were significantly decreased(P<0.01). Conclusion:Oxymatrine may alleviate ulcerative colitis by down-regulating the expression of Rho kinase,up-regulating the expressions of E-cadherin and TGF-β, inducing the apoptosis of intestinal epithelial cells, and mediating epithelial-mesenchymal transition.
