论文部分内容阅读
目的:探讨甲基苄基亚硝胺(MBNA)诱发食管癌前病变Wnt通路抑制因子的变化及膈下逐瘀汤的影响。方法:Wistar大鼠按3.5 mg·kg-1体重剂量皮下注射MBNA溶液,每周2次,造模首日起以膈下逐瘀汤16,8 g·kg-1剂量灌胃给药,于造模第10周处死各组大鼠,观察食管黏膜病理变化,荧光定量PCR检测sFRP1,sFRP4,Axin1,Axin2,GSK-3βmRNA转录水平,Western blot检测β-catenin蛋白水平。结果:MBNA诱导下,模型组大鼠食管黏膜病理学检查呈轻度不典型增生,膈下逐瘀汤高、低剂量组与模型组比较病理形态学与组织学均未有明显改善;模型组大鼠食管组织sFRP1,sFRP4,Axin1,Axin2基因转录水平较对照组降低(P<0.05或P<0.01),β-catenin蛋白水平较对照组升高(P<0.01),膈下逐瘀汤低剂量组sFRP4,Axin1,Axin2基因转录水平较模型组升高(P<0.05或P<0.01),β-catenin蛋白水平较模型组降低(P<0.01)。结论:上调β-catenin蛋白水平、下调Wnt通路抑制因子转录水平从而增强Wnt通路活性是MBNA致大鼠食管癌前病变的可能分子机制之一,膈下逐瘀汤虽可下调β-catenin蛋白水平、上调Wnt通路抑制因子转录水平,但不足以阻断MBNA诱发的食管癌前病变。
Objective: To investigate the changes of Wnt pathway inhibitor induced by methylbenzyl nitrosamine (MBNA) in esophageal precancerous lesions and the effect of Gexia Zhuyu Decoction. METHODS: Wistar rats were injected subcutaneously with 3.5 mg · kg-1 body weight of MBNA solution twice a week. The first day of modeling was given Gorgashi-Zhuyu Decoction at a dosage of 16,8 g · kg -1. Pathological changes of esophageal mucosa were observed at the 10th week after the model was established. The transcription levels of sFRP1, sFRP4, Axin1, Axin2 and GSK-3β were detected by real-time quantitative PCR and β-catenin protein level by Western blot. Results: The pathological changes of esophageal mucosa in model group were mild atypical hyperplasia induced by MBNA. The pathological morphology and histology of hypogastric stasis decoction high and low dose groups were not significantly improved compared with model group. The model group The transcription level of sFRP1, sFRP4, Axin1 and Axin2 in rat esophageal tissue was lower than that in control group (P <0.05 or P <0.01), while the protein level of β-catenin was higher in control group (P <0.01) The transcription level of sFRP4, Axin1 and Axin2 in the dose group was higher than that in the untreated group (P <0.05 or P <0.01), and the level of β-catenin was lower than that in the model group (P <0.01). CONCLUSION: Up-regulation of β-catenin protein level, down-regulation of Wnt pathway inhibitor transcription and enhancement of Wnt pathway activity are one of the possible molecular mechanisms of MBNA-induced esophageal precancerous lesions in rats. Gexiazhuyutang can down-regulate β-catenin protein level , Up-regulated Wnt pathway inhibitor transcription level, but not enough to block MBNA-induced esophageal precancerous lesions.