基于结肠黏膜ICAM-1探讨芍药汤对溃疡性结肠炎(胃肠湿热证)大鼠的作用机制

Discussion of the Mechanism of Peony Decoction on Gastrointestinal-syndrome Rats with Ulcerative Colitis Based on Colonic Mucosa ICAM-1

目的采用RT-PCR和Western-blot检测细胞间黏附分子-1(ICAM-1)基因生成以及蛋白的表达水平,探讨芍药汤治疗溃疡性结肠炎(UC)可能的作用机制。方法随机将48只清洁级SD大鼠分为4组,分别为空白组、模型组、西药对照组(SASP组)以及芍药汤组,每组12只;模型组、芍药汤组及SASP组3组大鼠在成功制成胃肠湿热证模型基础上,再建立UC模型;造模完成后,各组大鼠分别进行疾病活动指数(DAI)评分,灌胃治疗2周,处死大鼠前对其进行第2次DAI评分,完毕后麻醉并处死各组大鼠;解剖后,取病变的结肠组织观察,进行组织大体形态损伤(CMDI)评分;随后,将病变最明显结肠组织肠段2 cm左右,平均分为两等分,分别用于检测ICAM-1基因的蛋白的表达。结果相比空白组,模型组大鼠的DAI及CMDI评分升高明显(P<0.01),治疗后芍药汤组上述积分与模型组相比而言,下降明显(P<0.05);在结肠组织ICAM-1蛋白表达方面,Western-blot结果显示,模型组大鼠灰度值较空白组明显下降,治疗后,芍药汤组与SASP组ICAM-1灰度值均有所上升,且与正常组水平相近;大鼠结肠组织中ICAM-1 m RNA生成方面,与空白组相比而言,模型组上升明显(P<0.01),而在治疗后与模型组相比,芍药汤组及SASP组ICAM-1 m RNA生成水平显著降低(P<0.01)。结论芍药汤通过抑制UC(胃肠湿热证)大鼠结肠黏膜组织中ICAM-1基因及蛋白的表达,从而改善UC大鼠的症状及体征,这可能是其治疗UC的作用机制。

Objective：To study the feasible mechanism of Peony Decoction on ulcerative colitis by detecting the expression level of gene and protein of Intercellular adhesion molecule-1（ICAM-1） through RT-PCR and West-ern-blot. Methods： 48 clean grade SD rats were randomly divided into the blank group,the model group,Peony Decoction group and SASP group,12 rats in each. Rats in the model group,Peony Decoction group and SASP group were built with Gastrointestinal syndrome,and based on it,they were made into ulcerative colitis model. All rats were marked on DAI after modeling,and after 14 days＆#39; lavage treatment,they were marked on DAI for the second time. Then they were narcotized and killed,abnormal tissues with pathological changes were chosen to make CMDI scores;2 cm of each tissue was chosen to detect the expression level of gene and protein of ICAM-1. Results： Compared with the blank group,rats in the model group got a higher level in DAI and CMDI,and the difference was obvious （P〈0.01）. After treatment,DAI scores of rats in Peony Decoction group was lower than those in the model group （P〈0.05）. In the expression of protein of colonic mucosa ICAM-1,grey level of rats in the model group was significantly lower than that in the blank group. After treatment,it rose in SASP group and Peony Decoction group,close to that of the blank group. The expression of mRNA of colonic mucosa ICAM-1 in the model group was significantly higher than those in the blank group （P〈0.01）,and after treatment it dropped in SASP group and Peony Decoction group （P〈0.01）. Conclusion： Peony Decoction can obviously suppress the expression level of mRNA and protein of colonic mucosa ICAM-1,which may be the feasible mechanism of Pe-ony Decoction on Ulcerative colitis.