化痰消瘀方对胃癌前病变大鼠PTEN、FAK及paxillin表达的影响

Effects of Huatan Xiaoyu decoction on expressions of PTEN,FAK and paxillin in rats with precancerous lesions of gastric cancer

目的观察化痰消瘀方对胃癌前病变大鼠PTEN、FAK及paxillin表达的影响,从分子生物学水平探讨其逆转胃癌前病变的作用机制。方法选择90只4~5周龄SD雄性大鼠,随机取15只作为空白组,其余大鼠均采用N-甲基-N’-硝基N-亚硝基胍综合饥饱失常、浓盐水灌胃的方法制备胃癌前病变大鼠模型。将造模成功大鼠随机分为模型组,中药高、中、低剂量组及维酶素组,每组13只。空白组正常饮食,余均造模成功后,模型组给予生理盐水灌胃,中药高、中、低剂量组分别给予3 m L/kg、2 m L/kg、1 m L/kg化痰消瘀方灌胃,维酶素组给予10 m L/kg维酶素灌胃,均灌胃8周。采用HE染色法观察大鼠胃黏膜病理改变情况,应用免疫组化法检测胃黏膜组织中PTEN、FAK及paxillin的表达情况。结果 HE染色显示空白组大鼠胃黏膜无癌前病变改变,模型组均出现不同程度的胃癌前病变改变,中药高、中剂量组和维酶素组胃黏膜癌前病变情况均较模型组明显改善（P均〈0.05）,且中药高剂量组改善程度高于其他各给药组（P均〈0.05）。免疫组化结果显示PTEN在空白组强阳性表达;模型组鲜有表达;中药高、中、低剂量组表达量逐渐降低,但高于模型组（P均〈0.05）;中药高剂量组PTEN表达量高于其他各给药组（P均〈0.05）。FAK、paxillin在空白组极少表达,模型组表达量较空白组显著增加（P均〈0.05）;中药高、中、低剂量组二者表达量均明显低于模型组（P均〈0.05）,其中高剂量组表达量明显低于其他各给药组（P均〈0.05）。结论中药化痰消瘀方可显著改善胃癌前病变大鼠胃黏膜组织病理学情况,其作用机制可能是激活抑癌基因PTEN,调节FAK的去磷酸化,通过FAK/Src信号通路下调paxillin来诱导细胞凋亡。

Objective It is to observe the influence of Huatan Xiaoyu decoction （HTXYD） on expressions of PTEN, FAK and paxillin in rats with precancerous lesions of gastric cancer （PLGC） , and explore the mechanism of reversing the precan- cerous lesions of gastric cancer from the level of molecular biology. Methods Ninety male SD rats of 4 to 5-week-old were ran- domly divided into blank group, model group, high dose Chinese medicine group, medium dose Chinese medicine group, low dose Chinese medicine group, and Vitacoenzyme group. Except for blank group, all the other groups mainly used N-methyl- N＇-nitro-N-nitrosoguanidine （MNNG） , irregular diet and emotional stimuli to establish PLGC rat models. After successful modeling, the animals in the model group were given normal saline, and different dose HTXYD and Vitacoenzyme in the corre- sponding treat groups. 8 weeks later, the pathological changes of gastric mucosa was observed by HE staining, and the expres- sions of PTEN, FAK and paxillin was detected by immunohistochemistry. Results HE staining showed that there was no pre- cancerous lesion in the gastric mucosa of the rats in the blank group, but had different degree of precancerous lesion of gastric cancer in the model group. Compared with the model group, the gastric mucosal precancerous lesion of the high does and the middle dose Chinese medicine group and the Vitacoenzyme group were significantly improved （ all P 〈 0.05 ） ; and the im- provement of the high dose Chinese medicine group was better than that of the other treatment groups （ all P 〈 0.05 ）. The im- munohistoehemistry results showed that PTEN was strongly expressed in the blank group; there was little expression in model group ; the expressions of the high does, the medium does and the low dose Chinese medicine group were gradually decreased, but were significantly higher than that of the model group （ all P 〈 0.05） , and the expression of PTEN in the high dose group was significantly higher than that in the other treatment groups （ all P 〈 0.05）. Paxillin, FAK was very little expression in theblank group, but the expressions in the model group were significantly increased than that in the blank group （ all P 〈 0.05 ） ; their expression levels in the high does, the medium does and the low does Chinese medicine group were significantly lower than those in the model group （ all P 〈 0.05 ） , and which in the high dose Chinese medicine group was significantly lower than that in the other treatment groups （ all P 〈 0.05 ）. Conclusion HTXYD can significantly improve the pathological conditions of gastric mucosa to reverse the PLGC rats, the possible mechanism is to activate tumor suppressor gene PTEN, to regulate the dephosphorylation of FAK, and to down-regulate the paxillin through FAK/Src signaling pathway in aim to induce aooptosis.