[重组人转化生长因子-β1与重组人血小板源性生长因子-BB联合作用对SD大鼠正畸牙移动过程中Pyk2蛋白及mRNA基因表达的影响]

[Combined effect of rhTGF-β1 and rhPDGF-BB on the expression of Pyk2 protein and mRNA gene during orthodontic tooth movement in SD rats].

作者信息

Mei Mei, Zhang Jiang-Tao, Huang Jin, Feng Lei

机构信息

Department of Orthodontics, Affiliated Stomatology Hospital, Zunyi Medical College. Zunyi 563003, Guizhou Province, China. E-mail:

出版信息

Shanghai Kou Qiang Yi Xue. 2019 Oct;28(5):472-477.

PMID:32274476

Abstract

PURPOSE

To explore the combined effect of recombinant human transforming growth factor-β1 (rhTGF-β1) and recombinant human platelet derived growth factor-BB (rhPDGF-BB) on the expression of Pyk2 in the osteoclasts of the pressure side of orthodontic tooth in rats.

METHODS

One hundred and sixty male Sprague-Dawley rats were randomly divided into 2 groups: experimental group(A) and control group(B) to establish orthodontic tooth movement model. Rats in the experimental group received combined injection of 5 ng rhTGF-β1 and 10 ng rhPDGF-BB in the buccal submucosal area of the left upper first molar every other day, while rats in the control group received equivalent volumes of PBS. Rats in each group were sacrificed at each of 5 time points(1,4,7,10 and 14 days) after appliance placement. The distance of the tooth movement was measured with stereomicroscope, the change of the amount of osteoclasts was detected with tartrate-resistant acid phosphatase histochemistry (TRAP), the protein and mRNA expression of Pyk2 was detected by immunohistochemistry and real time PCR(RT-PCR). SPSS19.0 software package was used for statistical analysis.

RESULTS

The distance of teeth in group A moved more rapidly than in group B, except for the first day, significant differences were found (P<0.05) in all time points.The number of osteoclasts in group A was significantly higher than that in group B, except for the 14th day, compared with the control group(P<0.05). Pyk2 protein and gene expression of group A was significantly higher than group B, both groups reached a peak in 7 days and gradually decreased thereafter, except in the first day, the expression of protein Pyk2 was significantly different at other time points (P<0.05).

CONCLUSIONS

rhTGF-β1 and rhPDGF-BB up-regulated the expression of Pyk2 protein and mRNA gene on the pressure side of orthodontic tooth in rats, which may be one of the reasons for accelerating the rate of orthodontic tooth movement.

摘要

目的

探讨重组人转化生长因子-β1（rhTGF-β1）与重组人血小板衍生生长因子-BB（rhPDGF-BB）联合应用对大鼠正畸牙压力侧破骨细胞中Pyk2表达的影响。

方法

将160只雄性Sprague-Dawley大鼠随机分为2组：实验组（A组）和对照组（B组），建立正畸牙移动模型。实验组大鼠每隔一天在左上第一磨牙颊侧黏膜下区域联合注射5 ng rhTGF-β1和10 ng rhPDGF-BB，而对照组大鼠注射等量的PBS。每组大鼠在戴矫治器后的5个时间点（1、4、7、10和14天）分别处死。用体视显微镜测量牙齿移动距离，用抗酒石酸酸性磷酸酶组织化学法（TRAP）检测破骨细胞数量变化，用免疫组织化学和实时荧光定量PCR（RT-PCR）检测Pyk2的蛋白和mRNA表达。采用SPSS19.0软件包进行统计分析。

结果

除第一天外，A组牙齿移动距离比B组更快，各时间点差异均有统计学意义（P<0.05）。除第14天外，A组破骨细胞数量明显高于B组，与对照组相比差异有统计学意义（P<0.05）。A组Pyk2蛋白和基因表达明显高于B组，两组均在7天达到峰值，此后逐渐下降，除第一天外，其他时间点Pyk2蛋白表达差异均有统计学意义（P<0.05）。