叶酸对MNNG诱导的食管上皮细胞MAPK信号通路调控的干预研究  

Intervention of folic acid on regulation of MAPK signal pathway in esophageal epithelial cells induced by MNNG

作　　者：党兴波 韩菲 黄淑翠 匡增怡 甘秀凤 梁秋莲 张晓熙 张慧霞 DANG Xing-bo;HAN Fei;HUANG Shu-cui;KUANG Zeng-yi;GAN Xiu-feng;LIANG Qiu-lian;ZHANG Xiao-xi;ZHANG Hui-xia(Guilin Medical University,Guilin,Guangxi 541199,China)

机构地区：[1]桂林医学院,广西桂林541199

出　　处：《医药论坛杂志》2023年第8期1-5,共5页Journal of Medical Forum

基　　金：广西高校中青年教师科研基础能力提升项目(2019KY0546);桂林医学院大学生创新创业训练计划项目(201910601063)。

摘　　要：目的探究叶酸对N-甲基-N'-硝基-N-亚硝基胍(MNNG)诱导的人食管上皮细胞MAPK信号通路中关键因子表达情况的影响,为食管癌的营养防治提供理论依据。方法紫外分光光度法测定食物中亚硝酸盐的含量;采用Western Blot检测MAPK通路中关键因子ERK1/2、p38及其磷酸化蛋白表达情况。结果①抽取的桂林当地两市场米粉、酸笋、酸豆角中均含有少量亚硝酸盐,但其含量未超出国家标准;②经MNNG和不同浓度叶酸作用于人食管上皮细胞后,与对照组相比,除24h p38蛋白表达外,MNNG+叶酸中剂量组细胞ERK1/2、p38及其磷酸化蛋白表达水平均明显增高(t_(ERK1/2,12)=6.497,t_(ERK1/2,24)=18.873,t_(ERK1/2,48)=18.000,t_(p-ERK1/2,12)=82.452,t_(p-ERK1/2,24)=25.404,t_(p-ERK1/2,48)=28.206,t_(p38,12)=8.966,t_(p38,48)=22.808,t_(p-p38,12)=18.618,t_(p-p38,24)=44.156,t_(p-p38,48)=47.600;P<0.05);在作用不同时间时,随叶酸浓度增加细胞ERK1/2、p38及其磷酸化蛋白表达水平均降低(F_(ERK1/2,12)=26.892,F_(ERK1/2,24)=240.256,F_(ERK1/2,48)=26.210,F_(p-ERK1/2,12)=47.849,F_(p-ERK1/2,24)=318.176,F_(p-ERK1/2,48)=133.843,F_(p38,12)=4.533,F_(p38,24)=135.239,F_(p38,48)=156.389,F_(p-p38,12)=48.534,F_(p-p38,24)=3504.359,F_(p-p38,48)=115.241;P<0.05);在不同浓度叶酸作用,除中低剂量组ERK1/2蛋白表达外,不同时间组细胞ERK1/2、p38及其磷酸化蛋白表达水平有明显变化(F_(ERK1/2),高=165.046,F_(p-ERK1/2,低)=35.323,F_(p-ERK1/2),中=48.377,F_(p-ERK1/2,高)=8.083,F_(p38,低)=36.762,F_(p38,中)=60.923,F_(p38,高)=29.482,F_(p-p38,低)=169.324,F_(p-p38,中)=65.286,F_(p-p38,高)=29.256;P<0.05),低剂量组细胞各蛋白呈先增加后减少趋势,其余剂量组细胞呈现先减少后增加趋势。结论叶酸对MNNG诱导的人食管上皮细胞MAPK信号通路中关键因子ERK1/2、p38及其磷酸化蛋白表达具有一定的干预作用,在一定程度上对食管癌的防治提供了科学理论依据。Objective To explore the effect of folic acid on the expression of MAPK signaling pathway of human esophageal epithelial cells induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG),so as to provide theoretical basis for nutritional prevention and treatment of esophageal cancer.Methods The content of nitrite in characteristic food of Guilin was determined by ultraviolet spectrophotometry;By using Western Blot to detect the expression of key factors ERK1/2,p38 and their phosphorylated proteins in MAPK pathway.Results①The rice noodles,sour bamboo shoots and sour beans in the two local markets of Guilin contain a small amount of nitrite,which does not exceed the national standard.②After human esophageal epithelial cells were treated with MNNG and different concentrations of folic acid for different time,except for 24 h p38 protein expression,the expression levels of ERK1/2,p38 and their phosphorylated proteins in MNNG+folic acid medium dose group were significantly higher than those in the control group(t_(ERK1/2,12)=6.497,t_(ERK1/2,24)=18.873,t_(ERK1/2,48)=18.000,t_(p-ERK1/2,12)=82.452,t_(p-ERK1/2,24)=25.404,t_(p-ERK1/2,48)=28.206,t_(p38,12)=8.966,t_(p38,48)=22.808,t_(p-p38,12)=18.618,t_(p-p38,24)=44.156,t_(p-p38,48)=47.600;P<0.05);The expression levels of ERK1/2,p38 and their phosphorylated proteins decreased with the increase of folate concentration at different times(F_(ERK1/2,12)=26.892,F_(ERK1/2,24)=240.256,F_(ERK1/2,48)=26.210,F_(p-ERK1/2,12)=47.849,F_(p-ERK1/2,24)=318.176,F_(p-ERK1/2,48)=133.843,F_(p38,12)=4.533,F_(p38,24)=135.239,F_(p38,48)=156.389,F_(p-p38,12)=48.534,F_(p-p38,24)=3504.359,F_(p-p38,48)=115.241;P<0.05);In addition to the expression of E_(RK1/2) protein in the low-dose group,the expression levels of ERK1/2,p38 and their phosphorylated proteins changed significantly in different time groups(F_(ERK1/2,high)=165.046,F_(p-ERK1/2,low)=35.323,F_(p-ERK1/2,medium)=48.377,F_(p-ERK1/2,high)=8.083,F_(p38,low)=36.762,F_(p38,medium)=60.923,F_(p38,high)=29.482,F_(p-p38,low)=169.324,F_(p-p38,medium)=65.2

关 键 词：叶酸 食管癌 MNNG 信号通路 ERK1/2 p38 

分 类 号：R735.1[医药卫生—肿瘤]