浊毒1号方对胃癌前病变大鼠胃黏膜细胞凋亡及Bcl-2基因表达的影响
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摘要
目的:通过实验观察浊毒1号方对甲基硝基亚硝基胍(Methyl nitrate nitrosoguanidine,MNNG)溶液造模的胃癌前病变大鼠胃黏膜上皮细胞凋亡及Bcl-2基因表达的影响,以期从分子生物学角度探讨该方剂对胃癌前病变的影响。方法:将50只清洁级雄性Wista大鼠随机分为空白组、模型组、雷尼替丁组、中药高剂量组、中药低剂量组,每组各10只。除空白组外,其余4组用MNNG造模90天,大鼠造模成功后开始给予相应药物治疗4周后处死,检测各组大鼠胃黏膜中Tunel法标记的阳性颗粒,采用图像分析系统进行计数并进行统计学分析,检测胃黏膜细胞凋亡程度及癌基因中Bcl-2的表达水平。结果:与空白组比较,模型组大鼠胃黏膜上皮细胞凋亡及Bcl-2蛋白表达均上升(P <0.05)。与模型组比较,雷尼替丁组、中药高剂量组、中药低剂量组大鼠胃黏膜上皮细胞凋亡表达及Bcl-2蛋白表达均下降(P <0.05)。与雷尼替丁组比较,中药高剂量组凋亡细胞表达及Bcl-2蛋白表达均下降(P <0.05)。结论:浊毒1号方治疗胃癌前病变是通过下调Bcl-2基因水平,来减少胃黏膜细胞凋亡,从而恢复细胞正常生理结构的。
Objective:To observe the effect of Zhuodu No.1 prescription on gastric mucosal apoptosis and expression of Bcl-2 gene in the model of rats with gastric precancerous lesions which is established by methy lnitrate nitrosoguanidine(MNNG) solution via experiment so as to explore the effect of this prescription on gastric precancerous lesions from the perspective of molecular biology. Methods:50 male Wista rats of clean grade were divided into five groups randomly,namely the blank group,the model group,the ranitidine group and the high-dose and low-dose Chinese medicine groups,10 rats in each group. Except the blank group,the model of rats with gastric precancerous lesions was respectively established by MNNG in the other four groups for 90 days. After establishment, rats in each group received the corresponding medicine,which were sacrificed after 4 weeks of treatment. Detected the positive granules labeled by Tunel staining in gastric mucosa of rats in each group,conducted counting and statistically analysis via image analysis system,and detected the degree of gastric mucosal apoptosis and the expression of Bcl-2 in oncogene. Results: Comparing with the blank group, gastric mucosal apoptosis and the protein expression of Bcl-2 in oncogene of rats in the model group were both increased(P < 0.05).Comparing with the model group,gastric mucosal apoptosis and the protein expression of Bcl-2 in oncogene of rats in the ranitidine group and the high-dose and low-dose Chinese medicine groups were respectively decreased(P < 0.05). Comparing with the ranitidine group,gastric mucosal apoptosis and the protein expression of Bcl-2 in oncogene of rats in the high-dose Chinese medicine groups were both decreased(P < 0.05). Conclusion:Zhuodu No.1 prescription treats gastric precancerous lesions by down-regulating the expression of Bcl-2 gene to reduce gastric mucosal apoptosis,thus to promote the recovery of normal physiological structure of cells.
Objective:To observe the effect of Zhuodu No.1 prescription on gastric mucosal apoptosis and expression of Bcl-2 gene in the model of rats with gastric precancerous lesions which is established by methy lnitrate nitrosoguanidine(MNNG) solution via experiment so as to explore the effect of this prescription on gastric precancerous lesions from the perspective of molecular biology. Methods:50 male Wista rats of clean grade were divided into five groups randomly,namely the blank group,the model group,the ranitidine group and the high-dose and low-dose Chinese medicine groups,10 rats in each group. Except the blank group,the model of rats with gastric precancerous lesions was respectively established by MNNG in the other four groups for 90 days. After establishment, rats in each group received the corresponding medicine,which were sacrificed after 4 weeks of treatment. Detected the positive granules labeled by Tunel staining in gastric mucosa of rats in each group,conducted counting and statistically analysis via image analysis system,and detected the degree of gastric mucosal apoptosis and the expression of Bcl-2 in oncogene. Results: Comparing with the blank group, gastric mucosal apoptosis and the protein expression of Bcl-2 in oncogene of rats in the model group were both increased(P < 0.05).Comparing with the model group,gastric mucosal apoptosis and the protein expression of Bcl-2 in oncogene of rats in the ranitidine group and the high-dose and low-dose Chinese medicine groups were respectively decreased(P < 0.05). Comparing with the ranitidine group,gastric mucosal apoptosis and the protein expression of Bcl-2 in oncogene of rats in the high-dose Chinese medicine groups were both decreased(P < 0.05). Conclusion:Zhuodu No.1 prescription treats gastric precancerous lesions by down-regulating the expression of Bcl-2 gene to reduce gastric mucosal apoptosis,thus to promote the recovery of normal physiological structure of cells.
引文
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[9]原皓,李时光,崔慎茹.甘草酸二铵对溃疡性结肠炎大鼠结肠炎症及MCP-1表达的影响[J].山东医药,2010,50(31):41-42.
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[2]韦淑琴,刘国军,安洋,等.P53,nm23和c-erb B-2蛋白在子宫内膜癌中的表达及其临床意义[J].中国现代医学杂志,2000,10(10):9-11.
[3]徐婷婷,朱方石.外源性胃黏膜损伤因素与脾虚相关性的研究进展及评价[J].辽宁中医杂志,2015,37(6):1176-1179.
[4] Wallace JC,Granger DN.The cellular and molecular basis of gastric mucosal defense[J].FASEB J,1996,10(7):731-740.
[5] Krajewski S,Krajewska M,Shabaik A,et al. Immunohistochemical detemination of in vivo distribution of Bax,a dominant inhibitor of Bcl-2[J].Am J Pathol,1994,145:1323-1336.
[6] Hiwa H,Endo K,Wada R,et al.Celluar Proliferation and differentiation in rat atrophic gastic mucosa induced by N-methy-nitrosoguanidine[J].J clin Gasroenterol,1997:25:116-121.
[7]王正品,李佃贵,杜艳茹,等.浊毒致病论与现代中医病因学[J].中医杂志,2010,51(1):11-13.
[8]霍永利,李佃贵,马小顺.化浊解毒法治疗胃癌前病变患者61例[J].中医杂志,2011,52(8):698-699.
[9]原皓,李时光,崔慎茹.甘草酸二铵对溃疡性结肠炎大鼠结肠炎症及MCP-1表达的影响[J].山东医药,2010,50(31):41-42.
[10]徐新献,王志坦.中西医结合内科手册[M].成都:四川科学技术出版社,2003:383.
[11]沈敬华,杨丽敏,张林娜,等.5种中药提取物抗肿瘤作用的研究[J].内蒙古医学院学报,2005,27(4):300.