TGF-β2、TGF-β3启动子甲基化在TCDD诱发小鼠腭裂过程中的作用  

Role of TGF-β2,TGF-β3 promoter methylation in TCDD-induced cleft palate of mice

作　　者：陈露[1] 陈瑶 高湛[3] 刘小转 陶宇昌 余增丽 CHEN Lu;CHEN Yao;GAO Zhan;LIU Xiaozhuan;TAO Yuchang;YU Zengli(Department of Health Care,the Third Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Clinical Nutrition,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Gynecology and Obstetrics,the Fifth Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Single-cell Study Center,Henan Provincial People′s Hospital,Zhengzhou 450003;Department of Clinical Nutrition,Henan Provincial People′s Hospital,Zhengzhou 450003;Department of Nutrition and Food Hygiene,College of Public Health,Zhengzhou University,Zhengzhou 450001)

机构地区：[1]郑州大学第三附属医院保健部,郑州450052 [2]郑州大学第五附属医院临床营养科,郑州450052 [3]郑州大学第五附属医院妇产科,郑州450052 [4]河南省人民医院单细胞研究中心,郑州450003 [5]河南省人民医院临床营养科,郑州450003 [6]郑州大学公共卫生学院营养与食品卫生学教研室,郑州450001

出　　处：《郑州大学学报（医学版）》2022年第2期243-248,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(21577119,81801547)。

摘　　要：目的:探讨转化生长因子β(TGF-β)2、TGF-β3启动子甲基化模式在2,3,7,8-四氯二苯并对二噁英(TCDD)诱导腭裂发生中的作用。方法:将42只C57BL/6N孕鼠分为对照组和TCDD组,每组各21只。在妊娠第10天(GD10)暴露于64μg/kg TCDD建立腭裂小鼠模型,对照组采用等体积的玉米油灌胃。3~5 d后分别收集胎鼠头部用于后续检测。HE染色观察GD13~GD15胎鼠的腭部发育,取GD14胎鼠腭组织检测TGF-β2和TGF-β3启动子甲基化水平,利用qRT-PCR检测GD13~GD15胎鼠腭组织中TGF-β2、TGF-β3 mRNA的表达水平。结果:HE染色显示TCDD组胎鼠出现腭裂,而对照组未见腭发育异常。与对照组相比,TCDD组TGF-β2片段1的CpG10位点甲基化水平降低;TGF-β2片段2中以下CpG位点甲基化水平升高:CpG14、CpG18.19、CpG25和CpG29;而CpG32.33.34甲基化水平降低;TGF-β3片段1中,CpG2.3、CpG16.17.18位点均出现甲基化水平升高;TGF-β3片段2的CpG6甲基化水平升高,而CpG18甲基化水平降低(P<0.05)。与对照组相比,TCDD组GD13~GD15胎鼠腭组织中TGF-β2和TGF-β3 mRNA表达均降低(P<0.05)。结论:TCDD可能使TGF-β2和TGF-β3启动子部分位点发生甲基化而影响TGF-β2和TGF-β3在胎鼠腭部的表达,从而导致腭裂。Aim:To investigate the role of transforming growth factor-β(TGF-β)2,TGF-β3 promoter methylation in 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD)-induced cleft palate.Methods:Forty-two adult C57BL/6N pregnant mice were randomly allocated into control group(n=21)and TCDD group(n=21).The pregnant mice were exposed to 64μg/kg TCDD on gestation day 10(GD10)to establish cleft palate model,whereas the mice in the control group were given the equivalent volume of corn oil.Fetal rat heads were collected 3 to 5 days later for subsequent tests.The palatal development of GD13 to GD15 fetal mice was observed by HE staining.The GD14 palatal tissue was collected for the detection of TGF-β2 and TGF-β3 promoter methylation and the mRNA expression levels of TGF-β2 and TGF-β3 in the palatal tissue of GD13 to GD15 embryos were detected by qRT-PCR.Results:HE staining showed that cleft palate was observed in the TCDD group but not in the control group during GD13 to GD15.Compared with the control group,the CpG10 of TGF-β2 region-1 in TCDD group was significantly demethylated and in TGF-β2 region-2,CpG14,CpG18.19,CpG25,CpG29 were significantly methylated,while CpG32.33.34 were significantly demethylated(P<0.05).CpG2.3,CpG16.17.18 of TGF-β3 region-1 and CpG6 of TGF-β3 region-2 were significantly methylated,whereas in TGF-β3 region-2,CpG18 was significantly demethylated(P<0.05).Moreover,the mRNA expressions of TGF-β2 and TGF-β3 were significantly decreased in the TCDD group during GD13 to GD15(P<0.05).Conclusion:TCDD may affect the expressions of TGF-β2 and TGF-β3 in palate tissue of embryos during embryo mice palatogenesis by causing the methylation of some CpG sites in TGF-β2 and TGF-β3 promoter and thus leading to cleft palate.

关 键 词：表观遗传学 二噁英 腭裂 甲基化 转化生长因子β 小鼠 

分 类 号：R780.2[医药卫生—口腔医学]