还原型谷胱甘肽稳态在腭裂发生中的作用

doi:10.19438/j.cjoms.2018.03.005

中国口腔颌面外科杂志 ›› 2018, Vol. 16 ›› Issue (3): 215-221.doi: 10.19438/j.cjoms.2018.03.005

还原型谷胱甘肽稳态在腭裂发生中的作用

赵铜超，卞媛媛，张天奇，张东升，马利

山东大学附属省立医院口腔科，山东省口腔-头颈生物医学重点实验室，山东济南250012

收稿日期:2017-11-20 修回日期:2018-01-23 出版日期:2018-05-20 发布日期:2018-06-12
通讯作者: 马利，E-mail：mali3709@163.com
作者简介:赵铜超（1991-），男，在读博士研究生，E-mail：961497591@qq.com
基金资助:
国家自然科学基金（81401211）

Role of glutathione homeostasis on 2,3,7,8-tetrachlorodibenzo-p-dioxin induced cleft palates in mice

ZHAO Tong-chao, BIAN Yuan-yuan, ZHANG Tian-qi, ZHANG Dong-sheng, MA Li.

Department of Stomatology, Shandong Provincial Hospital Affiliated to Shandong University;Shandong Provincial Key Laboratory of Oral and Maxillofacial-Head and Neck Biomedicine. Jinan 250012, Shandong Province, China

Received:2017-11-20 Revised:2018-01-23 Online:2018-05-20 Published:2018-06-12

摘要/Abstract

摘要： 目的： 观察还原型谷胱甘肽稳态的改变对腭中嵴上皮细胞形态及腭裂发生率的影响，探讨氧化自由基在四氯二苯二噁英（TCDD）致畸过程中的作用。方法： 将GD10的SPF级C57BL/6J孕鼠随机分为4组，TCDD组：腹腔注射生理盐水和TCDD灌胃；TCDD+丁硫氨酸-亚砜胺(BSO)组：腹腔注射BSO 4 h后给予TCDD灌胃；BSO组：腹腔注射BSO和玉米油灌胃；对照组：腹腔注射生理盐水和玉米油灌胃。光学显微镜和电子扫描电镜下观察胎鼠腭胚突的发育及细胞表面形态。统计各组小鼠在GD17的腭裂发生率。免疫组织化学方法检测各组腭中嵴上皮各时期CYP1A1蛋白的表达。采用SPSS13.0软件包对数据进行统计分析。结果： TCDD成功构建高致畸率的腭裂模型。BSO增加了5%的腭裂发生率，但是相对于TCDD组无显著差异(P>0.05)。TCDD组腭胚突上抬时间较对照组推迟1 d，添加BSO的TCDD组腭胚突上抬过程较正常组减慢。电镜下胚胎发育的各发育时期，腭中嵴上皮表面TCDD组与对照组有显著差异。免疫组织化学检测发现，TCDD组腭中嵴上皮可见CYP1A1高表达。结论： 扰乱体内还原型谷胱甘肽稳态，可能会减少对小鼠体内氧化自由基的消耗，从而影响腭胚突的融合，继而加速了TCDD的毒性，影响腭裂发生。

关键词: TCDD, 腭裂, 还原型谷胱甘肽, BSO, CYP1A1

Abstract: PURPOSE: To elucidate the impact of glutathione homeostasis in organogenesis stage embryos on 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD) -induced cleft palate in fetal mice. METHODS: Pregnant C57BL/6J mice were randomly allocated to blank control group, TCDD exposed group, L-buthionine-sulfoximine (BSO)exposed group and BSO+TCDD exposed group. Each GD10 animal was weighted and treated with 0.1mL normal saline (for blank control group and TCDD exposed group) or 600 mg/kg BSO (for BSO group and TCDD+BSO group). After 4 hours, TCDD was orally administrated to subjects in both TCDD group and TCDD+BSO group, but corn oil was used as vehicle control for control group and BSO group. Fetal mice palates were imaged using light microscopy on GD13.5, GD14, GD14.25, GD14.5, GD14.75 and GD15; scanning electronmicroscopy was used on GD13.5, GD14.5, and GD15.5, and cleft palate were recorded on GD17.5. Three GD14.5 palate sections from each group were examined by immunohistochemistry.SPSS13.0 software package was used for statistical analysis. RESULTS: TCDD successfully induced cleft palate. BSO can increase 5% of cleft palate. In the TCDD treated group, palatal shelves elevated 1 day later than in the control group. In the TCDD and BSO group, 0.25 day later than the TCDD group, all palates had elevated by GD 15.0. After elevation, the shelves contacted each other and fused. Under a high magnification (×2500), compared to the control group (GD14.5), TCDD group (GD 15), no filaments were observed and cell shapes were flat with unclear boundaries. The group treated with TCDD and BSO had no significant difference from the TCDD group in the end, but had temporal variations. The TCDD exposed group had a higher expression of CYP1A1 on the frontal epithelia than the control group. CONCLUSIONS: Breaking the balance of glutathione may decline the formation of filopodia and MEE motility due to reducing the consumption of reactive oxygen species, filopodia and MEE movement. This may result in accelerating the ultrastructure change of the palatal surface to a normal state, leading to a higher rate of cleft palate in TCDD-treated fetal mice.

Key words: TCDD, Cleft palate, GSH, BSO, CYP1A1

中图分类号:

R782.2.2

赵铜超，卞媛媛，张天奇，张东升，马利. 还原型谷胱甘肽稳态在腭裂发生中的作用[J]. 中国口腔颌面外科杂志, 2018, 16(3): 215-221.

ZHAO Tong-chao, BIAN Yuan-yuan, ZHANG Tian-qi, ZHANG Dong-sheng, MA Li.. Role of glutathione homeostasis on 2,3,7,8-tetrachlorodibenzo-p-dioxin induced cleft palates in mice[J]. China J Oral Maxillofac Surg, 2018, 16(3): 215-221.

参考文献

[1] Abbott BD, Schmid JE, Brown JG, et al. RT-PCR quantification of AHR, ARNT, GR, and CYP1A1 mRNA in craniofacial tissues of embryonic mice exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin and hydrocortisone [J]. Toxicol Sci, 1999, 47(1): 76-85.
[2] Birnbaum LS. Developmental effects of dioxins and related endocrine disrupting chemicals[J]. Toxicol Lett, 1995, 82-83: 743-750.
[3] Gao Z, Bu Y, Liu X, et al. TCDD promoted EMT of hFPECs via AhR, which involved the activation of EGFR/ERK signaling [J]. Toxicol Appl Pharmacol, 2016, 298: 48-55.
[4] Chopra M, Schrenk D. Dioxin toxicity, aryl hydrocarbon receptor signaling, and apoptosis-persistent pollutants affect programmed cell death [J]. Crit Rev Toxicol, 2011, 41(4): 292-320.
[5] Harris C, Dixon M, Hansen JM. Glutathione depletion modulates methanol, formaldehyde and formate toxicity in cultured rat conceptuses [J]. Cell Biol Toxicol, 2004, 20(3): 133-145.
[6] Birnbaum LS, Harris MW, Miller CP, et al. Synergistic interaction of 2,3,7,8,-tetrachlorodibenzo-p-dioxin and hydrocortisone in the induction of cleft palate in mice [J]. Teratology, 1986, 33(1): 29-35.
[7] Pratt RM, Dencker L, Diewert VM. 2,3,7,8-Tetrachlorodibenzo-p-dioxin-induced cleft palate in the mouse: evidence for alterations in palatal shelf fusion [J]. Teratog Carcinog Mutagen, 1984, 4(5): 427-436.
[8] Yamada T, Mishima K, Fujiwara K, et al. Cleft lip and palate in mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin: a morphological in vivo study[J]. Congenit Anom (Kyoto), 2006, 46(1): 21-25.
[9] Fujiwara K, Yamada T, Mishima K, et al. Morphological and immunohistochemical studies on cleft palates induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice [J]. Congenit Anom (Kyoto), 2008, 48(2): 68-73.
[10] Wells PG, Lee CJ, McCallum GP, et al. Receptor- and reactive intermediate-mediated mechanisms of teratogenesis [J]. Handb Exp Pharmacol, 2010, (196): 131-162.
[11] Jang JY, Shin S, Choi BI, et al. Antiteratogenic effects of alpha-naphthoflavone on 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposed mice in utero [J]. Reprod Toxicol, 2007, 24(3-4): 303-309.
[12] Yuan X, He X, Zhang X, et al. Comparative study of folic acid and alpha-naphthoflavone on reducing TCDD-induced cleft palate in fetal mice[J]. Cleft Palate Craniofac J, 2017, 54(2): 216-222.
[13] Kawajiri K, Fujii-Kuriyama Y. Cytochrome P450 gene regulation and physiological functions mediated by the aryl hydrocarbon receptor [J]. Arch Biochem Biophs, 2007, 464(2): 207-12.
[14] Reichard JF, Dalton TP, Shertzer HG, et al. Induction of oxidative stress responses by dioxin and other ligands of the aryl hydrocarbon receptor [J]. Dose Response, 2006, 3(3): 306-331.
[15] Ilavarasi K, Chermakani P, Arif Nisha S, et al. Antioxidant compounds in the seaweed Gelidiella acerosa protects human peripheral blood mononuclear cells against TCDD induced toxicity [J]. Drug Chem Toxicol, 2015, 38(2): 133-144.
[16] Hassoun EA, Walter AC, Alsharif NZ, et al. Modulation of TCDD-induced fetotoxicity and oxidative stress in embryonic and placental tissues of C57BL/6J mice by vitamin E succinate and ellagic acid [J]. Toxicology, 1997, 124(1): 27-37.
[17] Yan J, Hales BF. Depletion of glutathione induces 4-hydroxynonenal protein adducts and hydroxyurea teratogenicity in the organogenesis stage mouse embryo [J]. J Pharmacol Exp Ther, 2006, 319(2): 613-621.
[18] Cuervo R, Covarrubias L. Death is the major fate of medial edge epithelial cells and the cause of basal lamina degradation during palatogenesis [J]. Development, 2004, 131(1): 15-24.

还原型谷胱甘肽稳态在腭裂发生中的作用

Role of glutathione homeostasis on 2,3,7,8-tetrachlorodibenzo-p-dioxin induced cleft palates in mice

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	高涛, 马莲, 罗奕, 孙勇刚. 鼻音计鼻音化率值与腭咽闭合状态关系的探讨[J]. 中国口腔颌面外科杂志, 2023, 21(4): 372-376.
[2]	蓝春慧, 张雪燕, 韦敏玲, 丁文炜, 梁建华. 整体性护理路径在先天性腭裂患儿手术护理中的应用[J]. 中国口腔颌面外科杂志, 2023, 21(3): 237-242.
[3]	褚珺, 胡明, 王国民, 吕挺正. 19例合并凝血功能异常的腭裂患儿围术期处理[J]. 中国口腔颌面外科杂志, 2022, 20(3): 282-286.
[4]	张凌, 仇琳, 纪均. 喷他佐辛预防儿童腭裂手术全麻苏醒期躁动的效果评价[J]. 中国口腔颌面外科杂志, 2021, 19(1): 40-44.
[5]	林晓锐, 李晨曦, 张秀华. 瑞芬太尼复合氯胺酮在婴幼儿唇腭裂手术中的麻醉效果分析[J]. 中国口腔颌面外科杂志, 2021, 19(1): 45-48.
[6]	赵欣然, 蔡鸣. 不同咽成形术式对腭裂术后继发腭咽闭合功能不全合并睡眠呼吸障碍的影响[J]. 中国口腔颌面外科杂志, 2021, 19(1): 76-81.
[7]	牟思璇, 朱妍菲, 杜常欣, 王雪纯, 吴艳棋, 梅鹏, 王旭东, 朱敏. 改良牙支持式上颌骨前部牵引成骨术与上颌骨Le Fort I型前徙术对唇腭裂患者腭咽部结构的影响[J]. 中国口腔颌面外科杂志, 2020, 18(5): 427-433.
[8]	张晓敏, 杨雅娴, 宗源, 林鸣. Noordhoff手术联合PNAM正畸治疗唇腭裂患儿的疗效评价[J]. 中国口腔颌面外科杂志, 2020, 18(4): 352-356.
[9]	张文静, 王霞. 非综合征唇裂伴或不伴腭裂对口腔健康相关生活质量的影响[J]. 中国口腔颌面外科杂志, 2020, 18(3): 255-259.
[10]	姚志涛, 安玮, 买买提吐逊·吐尔地. 50例唇腭裂患儿手术前、后血液生化指标及腭裂隙内骨再生评价[J]. 中国口腔颌面外科杂志, 2020, 18(1): 56-59.
[11]	阿力木江·苏甫尔, 比力克孜·玉素甫, 热娜古孜·阿不都米吉提. 新疆喀什地区非综合征型唇腭裂发病危险因素的病例对照研究[J]. 中国口腔颌面外科杂志, 2019, 17(5): 436-439.
[12]	石光, 张肖, 孙莹. PNAM正畸联合Noordhoff手术治疗唇腭裂患儿的效果及对腭部裂隙情况、感染发生率的影响[J]. 中国口腔颌面外科杂志, 2019, 17(4): 319-322.
[13]	张晓宇, 陈铮晰, 师佳君, 李竹君, 汤霞, 翁梦佳, 黄偲璟, 郑梦婷, 陈振琦. 400例唇腭裂患儿母乳喂养现况调查及影响因素分析[J]. 中国口腔颌面外科杂志, 2019, 17(4): 332-336.
[14]	吴忆来, 王国民, 杨育生, 万腾, 陈阳, 梁赟. 咽后壁组织瓣转移术后渗血的原因分析和处理[J]. 中国口腔颌面外科杂志, 2019, 17(2): 171-174.
[15]	马利, 石冰, 张东升, 郑谦. 细胞凋亡及上皮-间充质转化在腭胚突融合过程中的作用探讨[J]. 中国口腔颌面外科杂志, 2018, 16(6): 493-497.