中国实用口腔科杂志

中国实用口腔科杂志 ›› 2012, Vol. 5 ›› Issue (9): 527-532.

• 论著 • 上一篇    下一篇

局部转染人白细胞介素-10基因对去卵巢大鼠实验性牙周炎牙槽骨吸收的影响研究

陈群1，马守治1，郭建斌1，江俊1，赵欣1，闫福华1 ，肖殷2   

  1. 作者单位：1.福建省高校口腔医学重点实验室，福建医科大学口腔医学院，福州  350002；2.澳大利亚昆士兰科技大学，昆士兰  布里斯班  4701
  • 出版日期:2012-09-15 发布日期:2012-11-24
  • 基金资助:

    基金项目：福建省科技重点资助国际合作项目（2010I0006）；闽江学者奖励计划（闽教高〔2010〕104号）

Effect of local hIL-10 gene transfer on experimental periodontitis in ovariectomized rats 

CHEN Qun**，MA Shou-zhi， GUO　Jian-bin，JIANG　Jun，ZHAO　Xin，YAN Fu-hua， XIAO　Yin.   

  1.  *Key University Laboratory of Stomatology of Fujian Province and School of Stomatology，Fujian Medical University， Fuzhou 350002，China
  • Online:2012-09-15 Published:2012-11-24

摘要： 目的    观察局部注射人白细胞介素-10（hIL-10）质粒对去卵巢大鼠实验性牙周炎牙槽骨吸收的影响及其机制。方法    本研究于2009年1月至2011年3月在福建医科大学口腔医学院完成。将24只3月龄雌性SD大鼠随机分成4组，分别为假手术+hIL-10（SHAM+hIL-10）组、假手术+空载体（SHAM+VECTOR）组、去卵巢+hIL-10（OVX+hIL-10）组和去卵巢+空载体（OVX+VECTOR）组，每组6只。前两组大鼠实施卵巢切除假手术后12周，丝线结扎左侧上颌第二磨牙，制造实验性牙周炎模型（即形成SHAM+hIL-10+EP和SHAM+VECTOR+EP亚组），右侧不结扎作为对照牙（即形成SHAM+hIL-10+C和SHAM+VECTOR+C亚组）。后两组大鼠实施切除双侧卵巢手术后12周，丝线结扎左侧上颌第二磨牙，制造实验性牙周炎模型（即形成OVX +hIL-10+EP和OVX +VECTOR+EP亚组）。同时，在SHAM+hIL-10组大鼠的双侧上颌第二磨牙腭侧牙龈黏膜下和OVX+hIL-10组大鼠左侧上颌第二磨牙腭侧牙龈黏膜下注射hIL-10质粒（5μg）-脂质体（5μL）复合物；在SHAM+VECTOR组大鼠的双侧上颌第二磨牙腭侧牙龈黏膜下和OVX+VECTOR组大鼠的左侧上颌第二磨牙腭侧牙龈黏膜下注射空载体质粒（5μg）-脂质体（5μL）复合物。隔天注射1次，第7次注射后的48 h，处死大鼠。观察各组大鼠骨密度、血清生化指标、牙槽骨吸收和牙周组织细胞因子的变化。结果    转染hIL-10 质粒后，SHAM+hIL-10+C 组、SHAM+hIL-10+EP 组和 OVX+hIL-10+EP组根分叉区牙周膜IL-10 阳性细胞数目分别显著高于 SHAM+VECTOR+C 组、SHAM+VECTOR+EP 组 和 OVX+VECTOR+EP 组（均P＜0.05）。同SHAM+VECTOR+C 组比较，SHAM+hIL-10+C 组牙周膜 IL-1β 阳性细胞数目显 著 减 少 （P＜0.05） ， RANKL 阳 性 细 胞 数 目 显 著 增 加 （P＜0.05） 。 同SHAM+VECTOR+EP 组比较，SHAM+hIL-10+EP 组牙周膜的 IL-1β、IL-6、TNF-α 和 RANKL 阳性细胞数目均显著减少（P＜0.05）。同 OVX+VECTOR+EP组比较， OVX+hIL-10+EP 组牙周膜的 IL-1β、IL-6、RANKL 和 MMP-8 阳性细胞数目均显著减少（P＜0.05）。 结论    局部注射hIL-10质粒可抑制去卵巢大鼠实验性牙周炎的牙槽骨吸收，可能与牙周组织促炎因子的表达下降有关。

关键词: 牙周炎, 绝经后骨质疏松, 牙槽骨, 基因治疗, 细胞因子

Abstract: Objective    To investigate the effect of local hIL-10 gene transfer on bone metabolism， absorption of alveolar bone， and expressions of cytokines in periodontal tissue in ovariectomized rats. Methods    Twenty-four SD female rats were randomly divided into four groups. Group A： SHAM+hIL-10； Group B： SHAM+VECTOR； Group C： OVX+hIL-10； Group D： OVX+VECTOR. Twelve weeks after SHAM or OVX operation， the experimental periodontitis were induced by placing 4-0 silk ligatures around the cervix of the left upper second molars. At the same time， the complex of 5μg hIL-10 plasmid with 5μL liposome for Group A and C or 5μg vector plasmid with 5μL liposome for Group B and D were injected into the palatal gingiva of the upper second molars on left sides one time every two days. However， the palatal gingiva of the upper second molars on the right sides were also injected with the same complex as the left for Group A and Group B. Twenty hours after the seventh injection， the rats were sacrificed and bone density， serum biomechanical marker， absorption of alveolar bone and the numbers of cytokine-positive cells in the furcation were measured to detect the effects of hIL-10 gene therapy on bone metabolism， absorption of alveolar bone and the expression of cytokines in the periodontal tissue. Results    It was demonstrated that significantly increased hIL-10 expression was detected in the periodontal tissues after local gene delivery. It was also noted that after hIL-10 gene local delivery， the expression levels of  IL-1β， IL-6， TNF-α， RANKL and MMP-8 were significantly downregulated in the furcation of ligatured teeth and the alveolar bone absorption was significantly decreased （P < 0.05）. Conclusion    The hIL-10 gene transferred by local injection has significant effect on the absorption of alveolar bone and the expression of cytokines including IL-1β， IL-6， TNF-α， RANKL and MMP-8 in periodontal tissues of experimental periodontitis in ovariectomized rats.

Key words: periodontitis, osteoporosis after menapause, alveolar bone, gene therapy, cytokine

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