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[氟对大鼠切牙成釉细胞中MMP-20/TIMP-2表达的影响及褪黑素对氟中毒的拮抗作用]

[Effect of fluoride on the expression of MMP-20/TIMP-2 in ameloblast of rat incisor and the antagonistic effect of melatonin against fluorosis].

作者信息

Zhang Xue-li, Xi Shu-hua, Guo Xiao-ying, Cheng Guang-yan, Zhang Ying

机构信息

Department of Preventive Dentistry, School of Stomatology, China Medical University, Shenyang 110002 China.

出版信息

Shanghai Kou Qiang Yi Xue. 2011 Feb;20(1):10-5.

Abstract

PURPOSE

To study the effect of concentration of fluoride on the expression of matrix metalloproteinase-20(MMP-20) and tissue inhibitors of metalloproteinase-2 (TIMP-2) in the ameloblast of rat incisor,and explore the formation mechanism of dental fluorosis. By comparing the different expression of MMP-20,TIMP-2 between fluoride group and the melatonin group,to decide whether melatonin has antagonitic effect on dental fluorosis.

METHODS

Forty Wistar rats were randomly divided into 6 groups. The groups were as follows: control group,low-dose group, high-dose group,normal saline group and melatonin group. The animals were sacrificed 10 weeks after treatment. HE and immunohistochemical staining were used to observe the changes of ameloblasts and the expression of MMP-20 and TIMP-2 in rat incisors. MetaMorph microscope images analysis system was used to analyze the images, and SPSS12.0 software package was used for data analysis.

RESULTS

The surface of rat incisors fed with fluoride had chalky color change and cross stritations could be seen on the enamel surface.In the fluoride group,the ameloblasts were disarranged, cells arranged in multi-layer,even showing vacuolar change.The changes in the high-dose group was severer than the low-dose group. MMP-20, TIMP-2 were expressed both in the secretory ameloblasts, and in the odontoblasts.The expression of MMP-20 in rat's ameloblasts in the experimental group was significantly lower than that in the control group (P < 0.01); and no significant difference was found between the low-dose and high-dose groups(P > 0.05). The difference of expression of TIMP-2 was not significant among all the groups. The difference of expression of MMP-20 and TIMP-2 was not significant between the melatonin and the fluoride groups.

CONCLUSIONS

The excessive fluoride can inhibit the secretion of MMP-20 and disturb the balance between MMP-20 and TIMP-2,which lead to the delay of amelogenin removal and enamel demineralization. Melatonin has no antagonistic effect on the dental fluorosis. Supported by National Natural Science Foundation of China (30600509) and Natural Science Foundation of Liaoning Province (20102278).

摘要

目的

研究氟浓度对大鼠切牙成釉细胞基质金属蛋白酶20(MMP-20)和金属蛋白酶组织抑制剂2(TIMP-2)表达的影响,探讨氟斑牙的形成机制。通过比较氟组和褪黑素组MMP-20、TIMP-2表达的差异,判断褪黑素对氟斑牙是否具有拮抗作用。

方法

将40只Wistar大鼠随机分为6组,分别为对照组、低剂量组、高剂量组、生理盐水组和褪黑素组。处理10周后处死动物,采用苏木精-伊红(HE)染色和免疫组织化学染色观察大鼠切牙成釉细胞的变化及MMP-20和TIMP-2的表达。应用MetaMorph显微镜图像分析系统进行图像分析,采用SPSS12.0软件包进行数据分析。

结果

喂饲氟的大鼠切牙表面出现白垩色改变,釉质表面可见横纹。氟组成釉细胞排列紊乱,细胞多层排列,甚至出现空泡样改变,高剂量组改变较低剂量组严重。MMP-20、TIMP-2在分泌期成釉细胞及成牙本质细胞中均有表达。实验组大鼠成釉细胞中MMP-20的表达明显低于对照组(P<0.01),低剂量组与高剂量组之间差异无统计学意义(P>0.05)。各实验组TIMP-2表达差异无统计学意义。褪黑素组与氟组MMP-20和TIMP-2表达差异无统计学意义。

结论

过量氟可抑制MMP-20的分泌,破坏MMP-20与TIMP-2之间的平衡,导致釉原蛋白清除延迟和釉质脱矿。褪黑素对氟斑牙无拮抗作用。 本研究受中国国家自然科学基金(30600509)和辽宁省自然科学基金(20102278)资助。

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