【作者】 申载薰；

【导师】 曹志群；

【作者基本信息】 山东中医药大学 ， 中医内科学， 2011， 博士

【摘要】 目的：通过研究芪莲舒痞颗粒对CAG癌前病变大鼠胃黏膜组织端粒酶活性、端粒长度、DNA含量及NF-κB表达的影响,探讨芪莲舒痞颗粒逆转CAG癌前病变的作用机理。方法：采用‘’MNNG+雷尼替丁+乙醇+饥饱失调”的综合方法复制Wistar大鼠CAG癌前病变模型,按体重随机分为正常组、模型组、芪莲舒痞大、中、小剂量组和胃复春阳性对照组,分别给予药物干预后观察：免疫组化、TRAP-ELISA法检测端粒酶活性、端粒长度及NF-κB的表达,用流式细胞仪检测DNA含量。结果：①胃黏膜病理变化：造模各组大鼠胃黏膜炎症、萎缩、肠上皮化生、异型增生发生例数与正常组比较,均显著增加(P<0.01或P<0.05)；与模型组相比,QLSP大、中剂量组胃黏膜炎症发生例数均显著减少(P<0.01或P<0.05), QLSP大剂量组萎缩、肠上皮化生的发生例数显著减少(P<0.05)；药物干预各组间异型增生的发生例数无统计学差异。②胃黏膜端粒长度：正常组为20.01±1.40bp,模型组13.32±1.62bp,胃复春组16.91±1.88bp,QLSP大、中、小剂量组分别为19.27±1.94bp、17.91±1.63bp、16.23±2.16bp。经统计学分析,模型组、胃复春组、QLSP中、小剂量组端粒长度较正常组明显缩短(P<0.01)；经胃复春、QLSP治疗后,端粒长度较模型组均明显增长(P<0.01)。与胃复春组比较,QLSP大剂量组端粒长度显著增长(P<0.01),QLSP中、小剂量组则无明显差异。③胃黏膜端粒酶活性比较：正常组端粒酶阳性率为0(0/20),模型组为66.7%(10/15),胃复春组为33.3%(5/15),QLSP大、中、小剂量组分别为0(0/15)、26.7%(4/15)、33.3%(5/15)。经统计学分析,模型组、胃复春组、QLSP中小剂量组端粒酶活性与正常组比较明显增强(P<0.01或P<0.05)；QLSP大剂量组端粒酶活性较模型组明显降低,差异有统计学意义(P<0.05)。QLSP中剂量组端粒酶阳性率较胃复春组下降,但无统计学差异。④DNA含量比较：正常组为11.49±2.65μg,模型组36.48±4.81μg,胃复春组26.39±5.85μg,QLSP大、中、小剂量组分别为16.99±4.62μg、20.37±4.70μg、28.41±4.53μg。经统计学分析,模型组、胃复春组、QLSP大、中、小剂量组DNA含量较正常组明显增加(P<0.01)；经胃复春、QLSP治疗后,DNA含量较模型组明显减少(P<0.01)。与胃复春组比较,QLSP大、中剂量组DNA含量显著减少(P<0.01),QLSP小剂量组则无明显差异。⑤胃黏膜NF-κB的表达：正常组阳性表达率为0(0/20),模型组为80%(12/15),胃复春组为33.3%(5/15),QLSP大、中、小剂量组分别为26.7%(4/15)、40%(6/15)、60%(9/15)。经统计学分析,模型组、胃复春组、QLSP大、中、小剂量组胃黏膜NF-KB表达阳性表达率较正常组显著升高(P<0.01或P<0.05)；胃复春组和QLSP大剂量组胃黏膜NF-κB表达阳性表达率较模型组显著下降(P<0.01或P<0.05)；胃复春组、QLSP各剂量组胃黏膜NF-KB表达阳性表达率未见明显差异。结论：实验结果表明芪莲舒痞方能有效逆转CAG癌前病变,从而证明运脾益肾、化瘀解毒是治疗CAG癌前病变的有效方法。芪莲舒痞颗粒的作用机理可能与其抑制CAG癌前病变大鼠胃黏膜组织端粒长度的缩短,抑制端粒酶的活性及DNA含量的增加,降低DNA多倍体的比例,抑制NF-κB的表达等有关。芪莲舒痞颗粒可能从多个环节、多个靶点阻断和逆转胃黏膜癌前病变。更多还原

【Abstract】 Objective:To probe into the pharmacodynamic mechanism of Qilian Shupi granule by way of researching on its effecting the activity of telomerase, length of telomere and content of DNA, expression of NF-κB in Wistar rats.Methods:In experiments, models of precancerous lesions of gastric cancer were induced by cancerogenic agent in Wistar rats, which were divided randomly into six groups (normal group, model group, WeiFuchun group and big, middle and small dose QLSP group) according to their weights. After treated with QLSP and WeiFuchun for 12 weeks, main outcome measures:optical microscope was used to observe gastric mucous membrane; Immunohistochemical methods and TRAP-ELISA were used to monitor the expressions of activity of telomerase、length of telomere and expressions of NF-κB, the content of DNA was detected by FCM.Results:Animal experimental research showed that the happened number of gastric mucosal inflammation and atrophy, intestinal metaplasia, hyperplasia of gastric mucous membrane in built models groups was more than normal group, there were notable difference between them (P<0.01 or P<0.05); the length of telomere were increased in big and middle doses QLSP group, while they were increased slightly in Wei Fuchun group and lower greatly in model group, there were notable differences among them(P<0.01); The activity of telomerase were decreased in big and middle doses QLSP group, while they were decreased slightly in Wei Fuchun group and higher greatly in model group, there were notable differences among them(P<0.01). The content of DNA was decreased in big and middle doses QLSP group, while they were higher greatly in model group and decreased slightly in Wei Fuchun group, there were notable differences among them(P< 0.01). The expressions of NF-κB was inhibited in big doses QLSP group, while they were increased greatly in model group and inhibited slightly in Wei Fuchun group, there were notable differences among them (P<0.01).Conclusions:Splenogastric asthenia, accumulation of stagnant pathogenic factors in the body, and involvement of the kidney functions at the advanced stage are the main causes leading to the pathogenesis of precancerous lesions of chronic atrophic gastritis. Therefore, invigorating spleen and tonifying kidney, regulating qi to disperse stagnation, and dispersing accumulation of pathogenic factors to resolve masses are the principles in the management of the condition. The index from experimental research reveals its mechanism at the systemic, cellular and molecular levels in treating precancerous lesions of chronic atrophic gastritis. The QLSP recipe acts on many chains and targets of the disease to produce a comprehensive therapeutic effect, the therapeutic mechanism may have much to do with an overall effect of inhibition of expression of the activity of telomerase、length of telomere and content of DNA、expression of NF-κB. And thus the study enriched the knowledge of pathology and treatment of precancerous lesions of chronic atrophic gastritis.更多还原