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目的:观察溃疡性结肠炎相关癌变(ulcerative colitis associated carcinogenesis,UCAC,简称溃结相关癌变)小鼠组织形态、超微结构以及结肠黏膜β-catenin、T细胞因子4(T cell factor-4,TCF-4)的表达变化,探讨健脾清热活血方药防治溃结相关癌变的可能机制。方法:120只清洁级BALB/c小鼠随机分6组,每组20只,设正常组、模型组、美沙拉嗪0.4 g·kg-1阳性对照组、健脾清热活血方低、中、高剂量治疗组(4.29,8.58,17.16 g·kg-1);除外正常组,其余各组采用1,2-二甲肼(DMH)/葡聚糖硫酸钠(DSS)复合法制备溃结相关癌变模型,模型组予等剂量生理盐水,对照组予美沙拉嗪,治疗组予不同剂量健脾清热活血方药水提液,连续干预16周。应用光镜、透射电镜观察溃结相关癌变小鼠结肠组织形态学以及超微结构变化;免疫组化、Western-bolt及实时荧光定量PCR(Real-time PCR)检测结肠黏膜β-catenin、TCF-4蛋白及其mRNA表达变化。结果:模型组小鼠结肠光镜下可见黏膜上皮脱落,腺体萎缩、高级别上皮内瘤变,部分癌变,电镜下可见大、小不等、排列紊乱的上皮细胞,胞核不规则、核仁增大并上移;治疗组光镜下见黏膜充血肿胀,个别轻度不典型增生和淋巴滤泡出现,未见溃疡及癌变,电镜下可见上皮细胞体积明显变小,核染色质边集、核固缩或碎裂,核膜断裂,凋亡小体形成;与模型组比较,治疗组结肠黏膜损伤有不同程度改善(P<0.05)。模型组结肠β-catenin,TCF-4蛋白及其mRNA呈高表达特点;治疗组结肠β-catenin、TCF-4蛋白及其mRNA表达均呈下降趋势,与模型组比较,有统计学意义(P<0.05)。结论:健脾清热活血方药可能通过调控β-catenin,TCF-4表达,促进炎症缓解,发挥防治溃疡性结肠炎相关癌变的效应。
OBJECTIVE: To observe the morphology, ultrastructure and the expression of β-catenin, T cell factor-4 (TCF) in colorectal mucosa of ulcerative colitis associated carcinogenesis (UCAC) -4), to explore the possible mechanism of Jianpi Qingre Huoxue Recipe in preventing and treating cancerous lesions related to ulceration. Methods: A total of 120 clean BALB / c mice were randomly divided into 6 groups with 20 rats in each group. Normal control group, model group, Mesalazine 0.4 g · kg -1 positive control group, High dose treatment group (4.29,8.58,17.16 g · kg-1). Except for the normal group, the other groups were treated with 1, 2-dimethylhydrazine (DMH) / dextran sodium sulfate (DSS) The rats in the model group were given the same dose of normal saline and the control group were treated with mesalazine. The treatment group was given different doses of Jianpi Qingre Huoxue Recipe for 16 weeks. The morphological and ultrastructural changes of colonic mucosa were observed by light microscopy and transmission electron microscopy. The expressions of β-catenin and TCF-1 in colonic mucosa were detected by immunohistochemistry, Western-bolt and real-time PCR. 4 protein and its mRNA expression changes. Results: Mucous epithelial shedding, glandular atrophy, high grade intraepithelial neoplasia, and some cancerous lesions were found under the light microscope in colon of mice in model group. Epithelial cells with large and small irregular disordered arrangement were observed under electron microscope. Ren increased and moved up; the treatment group, mucosal hyperemia, some mild dysplasia and lymphoid follicles appeared in the light microscope, no ulceration and carcinogenesis. Under electron microscope, the volume of epithelial cells was significantly smaller, , Nuclear pyknosis or fragmentation, nuclear membrane rupture and apoptotic body formation. Compared with the model group, the colonic mucosal injury in the treatment group improved to some extent (P <0.05). The expression of β-catenin and TCF-4 protein in the model group were higher than those in the untreated group (P <0.05). The expressions of β-catenin and TCF-4 protein in the colon of the model group were significantly decreased <0.05). Conclusion: Jianpi Qingre Huoxue recipe may play an important role in prevention and treatment of ulcerative colitis-associated cancers by regulating the expression of β-catenin and TCF-4.