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目的:通过观察胃癌小鼠组织病理学、超微结构、P110、P-AKT、beclin-1及CDK1等指标变化,分析清热化湿方药防治胃癌的可能机制。方法:100只雄性Balb-c小鼠按体质量随机分为5组,即正常组、模型组及治疗组(低、中、高剂量)每组20只,应用Hp感染联合N-甲基-N-亚硝基脲自由饮水法诱导胃癌模型,第30周起分组给药即模型组予蒸馏水灌胃;治疗组予低、中、高剂量清热化湿方灌胃;连续10周。末次给药后处死全部小鼠,分离胃组织,予HE染色及透射电镜检测胃黏膜组织病理学及超微结构变化;予荧光定量PCR及免疫应迹法检测胃黏膜P110、P-AKT、beclin-1及CDK1表达。结果:模型组小鼠胃体、胃窦黏膜重度慢性炎症100%,不典型增生占65.3%,低分化腺癌57%;治疗组小鼠胃窦黏膜轻-中度慢性炎症,不典型增生占24%,未见癌变,以中、高剂量组更为明显。与模型组比较,有统计学意义(P<0.05)。电镜下见模型组上皮细胞微绒毛稀疏甚至脱落消失,细胞间复合连接松解消失,核固缩,异染色质块状分布于核膜下,细胞处于坏死期。治疗组可见上皮细胞体积明显变小,核染色质边集、核固缩,核膜断核碎裂;中、高剂量组见上皮细胞明显变小、空泡化,凋亡小体形成,可见自噬。模型组胃黏膜P110、P-AKT、beclin-1及CDK1蛋白及mRNA表达呈上升趋势,治疗组P110、PAKT、beclin-1及CDK1表达明显下降,有统计学意义(P<0.05)。结论:清热化湿方可能通过调控PI3K-AKT信号,诱导细胞自噬,下调beclin-1及CDK1表达而发挥防治胃癌的效用。
OBJECTIVE: To investigate the possible mechanism of Qingre Huashi Decoction in preventing and treating gastric cancer by observing the changes of histopathology, ultrastructure, P110, P-AKT, beclin-1 and CDK1 in mice with gastric cancer. Methods: One hundred male Balb-c mice were randomly divided into five groups according to body weight: 20 in normal group, model group and treatment group (low, middle and high dose). Hp infection combined with N-methyl- N-nitrosourea induced drinking water induction of gastric cancer model, the group from the 30th week administration of the model group was distilled water; the treatment group to low, medium and high doses of heat dampness prescription; continuous 10 weeks. After the last administration, all the mice were sacrificed and the gastric tissues were isolated. The pathological and ultrastructural changes of gastric mucosa were examined by HE staining and transmission electron microscopy. The expressions of P110, P-AKT, beclin -1 and CDK1 expression. Results: The gastric mucosa and antral mucosa of the model group had severe chronic inflammation of 100%, atypical hyperplasia of 65.3% and poorly differentiated adenocarcinoma of 57%. The mice in the treatment group had mild to moderate chronic inflammation of the gastric antrum and atypical hyperplasia 24%, no cancer, in the medium and high dose group more obvious. Compared with the model group, there was statistical significance (P <0.05). Under the electron microscope, the microvilli in the model group were sparse or even disappeared, and the lysis and disappearance of the compound connective cells disappeared. The nuclear pyknosis and heterochromatin lumps were distributed under the nuclear membrane, and the cells were in the necrosis stage. In the treatment group, the volume of epithelial cells was obviously smaller and the chromatin margination, pyknosis and nuclear fragmentation were observed. In the medium and high dose groups, epithelial cells were significantly smaller, vacuolated and apoptotic bodies were seen Autophagy. The expression of P110, P-AKT, beclin-1 and CDK1 protein and mRNA in gastric mucosa of model group showed an upward trend. The expressions of P110, PAKT, beclin-1 and CDK1 were significantly decreased in the model group (P <0.05). Conclusion: Qingre Huashi decoction may play a role in prevention and treatment of gastric cancer by regulating PI3K-AKT signal, inducing autophagy, down-regulating the expression of beclin-1 and CDK1.