健脾清热活血类方药介导β-catenin、C-myc表达干预溃疡性结肠炎相关癌变研究

The intervention effect of Jianpi Qingre Huoxue Decoction on the expression of β-catenin and C-myc in ulcerative colitis associated carcinogenesis

目的探讨溃疡性结肠炎相关癌变结肠黏膜β-catenin、C-myc表达变化以及健脾清热活血方(救必应、白术、白芍、水蛭、三七、炙甘草)干预溃结癌变的机制。方法 60只清洁级Balb/c小鼠随机分5组,每组12只,正常组、模型组、低、中、高剂量治疗组;除正常组外,其余各组采用DMH/DSS复合法制备溃结癌变模型,模型组给予等剂量生理盐水、治疗组分别给予不同剂量健脾清热活血方药提取物干预20周。分别应用光镜检测结肠黏膜组织形态学变化,免疫组化及实时荧光定量PCR检测结肠黏膜β-catenin、C-myc蛋白及其mRNA表达变化。结果模型组小鼠结肠肉眼可见浆膜层散在、多处息肉状隆起变化,镜下见腺体萎缩、高级别上皮内瘤变,部分癌变,其结肠β-catenin、C-myc蛋白及其mRNA呈高表达特点;低剂量组肉眼见结肠黏膜充血水肿,部分可见溃疡样改变,镜下见黏膜充血肿胀,部分腺体萎缩,低级别上皮内瘤变,结肠β-catenin、C-myc蛋白及其mRNA表达呈下降趋势,与模型组比较,有统计学意义(P<0.05);中、高剂量组肉眼见结肠黏膜充血、水肿,糜烂,镜下见黏膜上皮脱落,充血肿胀,未见溃疡及癌变,结肠β-catenin、C-myc蛋白及其mRNA表达呈下降趋势,与模型组比较,有统计学意义(P<0.05);各治疗组比较,β-catenin、C-myc变化无统计学意义(P>0.05)。结论健脾清热活血方药可能通过介导β-catenin、C-myc表达,发挥防治溃结相关癌变的效应。

AIM To explore the mechanism of Jianpi Qingre Huoxue Decoction（Ilicis rotundae Cortex,Atractylodis macrocephale Rhizoma,Paeoniae Radix alba,Hriudo,Notoginseng Radix et Rhizoma,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle） for preventing ulcerative colitis associated with carcinogenesis（UCAC）,observing the changes of β-catenin and C-myc in colon mucosa.METHODS Sixty Balb/c rats were randomly divided into five groups,including normal group,blank group,intervention group（low dosage,medium dosage,high dosage） with each group of twelve rats.Except normal group,the other groups were intraperitoneally injected with 0.4% DMH/DSS for two weeks.The blank group was treated with normal saline and those intervention groups were treated with different dosage of Jianpi Qingre Huoxue Decoction for twenty weeks.The colon histomorphology was assayed by naked eyes and light microscope.The expressions of β-catenin and C-myc were detected by immunohistochemistry and Real-time PCR.RESULTS There were changes of hyperemia and edema and polyps occupation in colon mucosa by naked eyes.At the same time,the changes of high-grade intraepithelial neoplasia and carcinogenesis were seen under microscope with β-catenin and C-myc over-expression in blank group,compared with the intervention group（P 0.05）.However,the histomorphologic changes in low dosage intervention group was characterized by hyperaemia and ulcer and low-grade intraepithelial neoplasia,and in the medium-high dosage group was characterized by hyperaemia and edema.The expressions of β-catenin and C-myc in intervention groups were on the decline（P〈0.05）.CONCLUSION Jianpi Qingre Huoxue Decoction may exert a preventing effect on ulcerative colitis associated with carcinogenesis through mediating the expressions of β-catenin and C-myc.