消痔丸对醋酸致肛门溃疡模型大鼠的作用机制研究

Mechanism of Xiaozhi Pills on acetic acid-induced anal ulcer model rats

目的探讨消痔丸对醋酸诱导大鼠肛门溃疡模型的药效作用及其可能的作用机制。方法以醋酸诱导大鼠肛门溃疡模型,将造模成功大鼠分为模型组、消痔丸(0.81、1.62、3.24 g/kg)组、柑橘黄酮片(0.27 g/kg)组和痔炎消片(0.86 g/kg)组,另设置对照组,每组10只。对照组与模型组ig给予同等体积的0.1%CMC-Na溶液。各组连续ig给药10 d,1次/d。观察各组大鼠表观指标,并采用Image J软件计算溃疡面积;苏木素–伊红染色法观察肛门直肠组织病理学,并进行分级评分;酶联免疫吸附法(ELISA)检测大鼠肛门组织中基质金属蛋白酶-9(MMP-9)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)含量;免疫组化法检测肛门组织中诱导型一氧化氮合成酶(iNOS)、转化生长因子β1(TGFβ1)、血管内皮生长因子A(VEGFA)蛋白表达。结果与模型组相比,消痔丸组可显著改善醋酸所致大鼠肛周溃疡、肿胀、黏液分泌等情况,加快大鼠肛周黏膜上皮修复,溃疡面积缩小,减少间质或黏膜下水肿及炎性细胞浸润和炎性渗出物;消痔丸组可抑制肛周组织中MMP-9、IL-6、TNF-α分泌和iNOS、VEGFA蛋白表达,增加TGF-β1表达(P<0.05、0.001)。结论消痔丸对醋酸诱导的肛门溃疡模型治疗作用显著,主要通过抑制IL-6、TNF-α、MMP-9相关细胞因子分泌和iNOS、VEGFA蛋白表达,升高TGF-β1表达,从抗炎消肿、抑制基质降解、促进组织生长因子表达等途径,起到消肿生肌作用。

Objective To explore the pharmacodynamic effect and possible mechanism of Xiaozhi Pills on acetic acid induced anal ulceration rat model.Methods Rats with acetic acid-induced anal ulcer model were divided into model group,Xiaozhi Pills(0.81,1.62,3.24 g/kg)group,Citrus Bioflavonoids Tablets 0.27 g/kg group and Zhiyanxiao Tablets 0.86 g/kg group,and control group with 10 rats in each group.Control group and model group were given the same volume of 0.1%CMC-Na solution by gavage.Each group was given ig for 10 days,once daily.The apparent indexes of rats in each group were observed,and the ulcer area was calculated by Image J software.The histopathology of anus and rectum was observed by hematoxylin-eosin staining and graded.The contents of matrix metalloproteinase-9(MMP-9),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in rat anal tissue were detected by enzyme-linked immunosorbent assay(ELISA).The expression of inducible nitric oxide synthase(iNOS),transforming growth factorβ1(TGFβ1)and vascular endothelial growth factor A(VEGFA)protein in anal tissues was detected by immunohistochemistry.Results Compared with the model group,Xiaozhi Pills can significantly improve the perianal ulcer,swelling,and mucus secretion induced by acetic acid,accelerate the repair of perianal mucosal epithelium,and reduce interstitial or submucosal edema,inflammatory cell infiltration and inflammatory exudate.Xiaozhi Pills group could inhibit the secretion of MMP-9,IL-6,TNF-αand the protein expression of iNOS and VEGFA in perianal tissue,and increase the expression of TGF-β1(P<0.05,0.001).Conclusion Xiaozhi Pills has a significant therapeutic effect on acetic acid induced anal ulcer model,mainly by inhibiting the secretion of IL-6,TNF-α,MMP-9 related cytokines and the expression of iNOS and VEGFA protein,increasing the expression of TGF-β1,and playing a role in reducing swelling and promoting muscle growth through anti-inflammation and detumification,inhibiting matrix degradation,and promoting the expression of tissue growth factor.