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论文题名(中文):

 氟中毒对动物牙齿中COLⅠ和DPP表达的影响

作者:

 韩天龙

论文语种:

 chi

学科名称:

 临床兽医学

学位:

 农学硕士

学校:

 山西农业大学

院系:

 动物科技学院

专业:

 临床兽医学

研究方向:

 畜禽营养代谢与中毒病

第一导师姓名:

 王俊东

论文完成日期:

 2008-04-20

论文题名(外文):

 Effect of fluoride on the expression of Collagen typeⅠ and Dentin phosphoprotein in tooth

关键词(中文):

  门齿 臼齿 胶原蛋白 牙本质磷蛋白 绵羊 豚鼠

关键词(外文):

 Fluoride Incisor Molar Collagen typeⅠ Dentin phosphoprotein Sheep Guinea pig.

论文文摘(中文):
摘要
[目的] 研究自然病例和实验性氟中毒对牙齿中Ⅰ型胶原蛋白和牙本质磷蛋白表达
的影响,为从分子生物学水平揭示氟斑牙的形成机理提供依据。
[方法] ⑴ 选取经诊断为氟斑牙的3 周岁左右内蒙古裘皮型小尾寒羊雌性4 只作为
实验组;另选同一品种年龄相似的健康绵羊4 只作为对照组。分别取门齿和臼齿。
⑵ 选购Hartley 白色雄性豚鼠20 只,按照体重随机分为2 组,每组10 只,即对
照组和加氟组,分别自由饮用去离子水和150mg/L NaF 去离子水。所有动物均采食正常
标准豚鼠颗粒饲料,饲养周期为90 天。取门齿和臼齿。
将上述采集的绵羊和豚鼠的牙齿,经HE 和Masson 三色染色观察组织病理切片;用
免疫组化检测Ⅰ型胶原蛋白和牙本质磷蛋白的表达;并用一步法从中提取总RNA,设计
特异性引物,采用QRT-PCR 技术定量测定COL1A1 和DSPP 基因的表达。
[结果] ⑴ 绵羊及豚鼠与对照组相比,氟中毒组牙齿呈现波状齿及氟斑牙,牙本质
胶原排列紊乱,结构疏松,釉质及牙髓均出现损伤,牙髓成牙本质细胞萎缩,排列紊乱。
⑵ 与对照组相比,中毒组绵羊门齿和臼齿牙本质COLⅠ表达量均降低,差异极显著
(p<0.01);对照组绵羊牙本质COLⅠ表达量门齿低于臼齿,差异极显著(p<0.01);中毒
组绵羊牙本质COLⅠ表达量门齿低于臼齿,差异显著(p<0.05)。
⑶ 与对照组相比,中毒组绵羊门齿牙本质DPP 表达量降低,差异极显著(p<0.01);
中毒组绵羊臼齿牙本质DPP 表达量降低,差异显著(p<0.05)。对照组绵羊牙本质DPP 表
达量门齿低于臼齿,但无显著性差异;中毒组绵羊牙本质DPP 表达量门齿低于臼齿,差
异显著(p<0.05)。
⑷ 与对照组相比,中毒组豚鼠门齿和臼齿牙本质COLⅠ表达量均降低,差异极显著
(p<0.01);对照组和中毒组内,豚鼠牙本质COLⅠ表达量门齿均低于臼齿,差异极显著
(p<0.01)。
⑸ 与对照组相比,中毒组豚鼠门齿和臼齿牙本质DPP 表达量均降低,差异极显著
(p<0.01)。对照组和中毒组内,豚鼠门齿牙本质DPP 表达量与臼齿无显著性差异。
⑹ 绵羊氟中毒组门齿和臼齿COL1A1 基因的相对表达量均低于对照组;对照组和中
毒组内,门齿COL1A1 基因的相对表达量低于臼齿。
⑺ 绵羊氟中毒组门齿和臼齿DSPP 基因的相对表达量均低于对照组;对照组和中毒
组内,门齿DSPP 基因的相对表达量低于臼齿。
⑻ 豚鼠氟中毒组门齿和臼齿COL1A1 基因的相对表达量均低于对照组;对照组和中
毒组内,门齿COL1A1 基因的相对表达量低于臼齿。
⑼ 豚鼠氟中毒组门齿和臼齿DSPP 基因的相对表达量均低于对照组;对照组和中毒
组内,门齿DSPP 基因的相对表达量低于臼齿。
[结论] ⑴ 染毒90 天150mg/L NaF 的Hartley 白色雄性豚鼠,可造成自然状态下
慢性氟中毒的波状齿及氟斑牙模型。
⑵ 工业氟污染可影响内蒙古雌性裘皮型小尾寒羊门齿和臼齿中COLⅠ和DPP 的表
达,导致COLⅠ及其COL1A1 基因、DPP 及其DSPP 基因在门齿和臼齿中的表达均降低。
⑶ 150mg/L NaF 可影响Hartley 白色雄性豚鼠门齿和臼齿中COLⅠ和DPP 的表达,
导致COLⅠ及其COL1A1 基因、DPP 及其DSPP 基因在门齿和臼齿中的表达均降低。
⑷ 工业氟污染对绵羊臼齿的损伤及对COLⅠ及其COL1A1 基因表达的抑制较门齿严
重。
文摘(外文):
ABSTRACT
[Objective] We studied the effect of fluoride on the expression of COLⅠ and DPP in tooth, to explore the mechanism of fluorosis from the level of molecular biology, and to provide the theoretical foundation for mottled enamel.
[Method] (1) Chose four female fluorosed Inner Mongolia small fattail cold sheep aged three years for fluorosed group, and four healthy sheep with the same age, breed and gender for control. Collected their incisors and molars. (2) Twenty male Hartley albino guinea pigs were randomly divided into two groups of ten, according to the avoirdupois.The control group drunk doubledistilled water and the fluorosed group
drunk doubledistilled water added 150mg/L NaF. All the animals were fed with the same diets for ninety days. Collected their incisors and molars. All the teeth were applied for the HE staining, Masson staining, Immunohistochemical staining (IHC) and realtime fluorescence quantitative reverse transcription polymerase chain reaction (QRTPCR) to observe the pathological damage, and to determine the expression of COLⅠ protein, DPP protein and COL1A1 gene, DSPP gene.
[Result] (1) Both in sheep and guinea pigs: compared with the control group, the teeth in the fluorosed group appeared mottled enamel (incisor) and waveformed mouth (molar)? the collagens in dentine row loosely and disorderly? the enamel and endodontics were damaged? the dentinoblasts shrunk and row loosely. (2) In sheep: compared with the control group, the expression of COLⅠ was obviously lower both in fluorosed incisor and molar (p<0.01)? in the control group, the expression of COLⅠ in incisor was obviously lower than in molar (p<0.01)? in the fluorosed group, the expression of COLⅠ in incisor was
lower than in molar too (p<0.05). (3) In sheep? compared with the control group, the expression of DPP was obviously lower in fluorosed incisor (p<0.01), the expression of DPP was lower in fluorosed molar (p<0.05)? in the control group, the expression of DPP in incisor was lower than in molar, but there was no significant difference (p>0.05)? in the fluorosed group, the expression of DPP in incisor was lower than in molar (p<0.05). (4) In guinea pigs: compared with the control group, the expression of COLⅠ was obviously lower both in fluorosed incisor and molar (p<0.01)? both in the control group and fluorosed group, the expression of COLⅠ in incisor was obviously lower than in molar (p<0.01). ⑸ In guinea pigs:
compared with the control group, the expression of DPP was obviously lower both in fluorosed incisor and molar (p<0.01)? both in the control group and fluorosed group, there was no significant differences in the expression of DPP between incisor and molar (p>0.05). ⑹ In sheep: compared with the control group, the expression of COL1A1 gene was lower both in fluorosed incisor and molar? both in the control group and
fluorosed group, the expression of COL1A1 gene in incisor was lower than in molar. ⑺ In sheep:
compared with the control group, the expression of DSPP gene was lower both in fluorosed incisor and molar? both in the control group and fluorosed group, the expression of DSPP gene in incisor was lower than in molar. ⑻ In guinea pigs:compared with the control group, the expression of COL1A1 gene was lower both in fluorosed incisor and molar? both in the control group and fluorosed group, the expression of COL1A1 gene in incisor was lower than in molar. ⑼ In guinea pigs: compared with the control group, the expression of DSPP gene was lower both in fluorosed incisor and molar? both in the control group and
fluorosed group, the expression of DSPP gene in incisor was lower than in molar.
[Conclusion] ⑴ Drunk doubledistilled water added 150mg/L NaF in guinea pig for ninety days, we made the natural and chronic fluorosis model successfully with mottled enamel and waveformed mouth. ⑵ Industrial fluoride pollution can affect the expression of COLⅠ and DPP in incisor and molar of Inner Mongolia small fattail cold sheep, inducing lower expression of COL1A1 gene and COLⅠ, and lower expression of DSPP gene and DPP both in incisor and molar. ⑶ 150mg/L NaF can affect the expression of COLⅠ and DPP in incisor and molar of Hartley albino guinea pigs, inducing lower expression of COL1A1
gene and COLⅠ, and lower expression of DSPP gene and DPP both in incisor and molar. ⑷ The damage of industrial fluoride pollution in the molar of sheep and the restrain of industrial fluoride pollution on the expression of COL1A1 gene and COLⅠ were more severely than in incisor.
开放日期:

 2009-04-20

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