解毒化瘀健脾方对胃黏膜异型增生模型大鼠p16、PTEN基因的去甲基化和蛋白诱导表达  

Jiedu Huayu Jianpi Fang induces demethylation and increased expression of p16 and PTEN genes in gastric dysplasia in rats

作　　者：李志钢 张伟 邱作成 夏宽宏 纪勇[4] 李玲[4] 连军[4] 安娟[4] 

机构地区：[1]新疆维吾尔自治区中医医院专家门诊,新疆维吾尔自治区乌鲁木齐市830000 [2]新疆维吾尔自治区中医医院耳鼻喉科,新疆维吾尔自治区乌鲁木齐市830000 [3]新疆维吾尔自治区中医医院病理科,新疆维吾尔自治区乌鲁木齐市830000 [4]新疆医科大学动物实验中心,新疆维吾尔自治区乌鲁木齐市830000

出　　处：《世界华人消化杂志》2014年第9期1247-1255,共9页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81060288~~

摘　　要：目的:观察解毒化瘀健脾方对胃黏膜异型增生模型大鼠细胞周期蛋白依赖性激酶抑制剂2A(cyclin-dependent kinase inhibitor 2A,p16)、磷酸酶张力蛋白同源蛋白(phosphatase and tensin homolog,PTEN)基因甲基化状态和蛋白表达的影响,并探讨解毒化瘀健脾方对胃黏膜异型增生的治疗作用.方法:采用低浓度N-甲基-N'-硝基-N-亚硝基胍(N-methyl-N'-nitro-N-nitrosoguanidine,MNNG)的综合造模方法,创建实验性大鼠胃黏膜异型增生病变模型;分模型对照组、西药维甲酸治疗组、解毒化瘀健脾方治疗组,并选择正常大鼠作为阳性对照组进行干预;应用甲基化特异PCR技术检测大鼠胃黏膜p16、P T E N基因甲基化状态;R e a l-t i m e P C R、Western blot、免疫组织化学技术检测p16,PTEN在mRNA和蛋白表达水平的变化.结果:模型组胃黏膜异型增生细胞p16、PTEN基因的甲基化阳性检出率均为33.33%(6/18),均高于正常对照组20%(2/10),但差异不显著;解毒化瘀健脾方治疗组p16基因甲基化检出率均未0%(0/15),相比模型组20%(2/10)甲基化程度降低,但差异均无统计学意义;p16在mRNA(P<0.001)和蛋白(P<0.01)水平的表达相比模型组极显著升高;PTEN在mRNA(P<0.05)和蛋白(P<0.01)水平的表达相比模型组极显著升高.结论:解毒化瘀健脾方对异型增生胃黏膜细胞p16、PTEN基因具有一定的去甲基化作用,并显著诱导二者的表达量增加.解毒化瘀健脾组方的对胃黏膜异型增生具有潜在的治疗作用.AIM： To observe the effect of Jiedu Huayu Ji- anpi Fang （JHJF） on the methylation status and expression of cyclin-dependent kinase inhibitor 2A （p16） and phosphatase and tensin homolog （PTEN） in rats with gastric dysplasia （GD）, and to explore the therapeutic effect of JHJF on GD. METHODS： A low concentration of N-methyl -N＇-nitro-N-nitrosoguanidine （MNNG）-based modeling method was adopted to induce GMD, and model rats were randomly divided into a model control group, a retinoic acid treatment group, and a JHJF treatment group. Normal rats treated with JHJF were used as positive controls. Methylation specific PCR was used to detect the methylation status of p16 and PTEN genes in gastric mucosal ceils, and real-time PCR, West- ern blot and immunohistochemistry were used to detect the mRNA and protein expression of p16 and PTEN. RESULTS： The rates of p16 and PTEN gene methylation in the model control group were both 33.33% （6/18）, higher than those in the normal control group 20% （2/10）, but the differ- ences were not significant. In the JHJF treatment group, the rates of p16 and PTEN gene methyla- tion were both 0% （0/15）, significantly lower than those in the model control group （33.33%, 6/18）. The expression levels of p16 and PTEN mRNAs （P 〈 0.001, P 〈 0.05） and proteins （P 〈 0.01 for both） were significantly higher in the JHJF treatment group than in the model control groups. CONCLUSION： JHJF can induce demethylation and increase expression of the p16 and PTEN genes in GD in rats. JHJF has a potential value for the treatment of GD.

关 键 词：解毒化瘀健脾 胃黏膜异型增生 P16基因 PTEN基因 甲基化 

分 类 号：R273[医药卫生—中西医结合]