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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48h,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。
Objective To investigate the effect and mechanism of local injection of human interleukin-10 (hIL-10) on alveolar bone in experimental periodontitis in ovariectomized rats. Methods The study was performed at the School of Stomatology, Fujian Medical University from January 2009 to March 2011. Twenty-four three-month-old female Sprague-Dawley rats were randomly divided into 4 groups: sham operated + hIL-10 group, SHAM + VECTOR group, ovariectomized + hIL- (OVX + hIL-10) group and ovariectomized + empty vector (OVX + VECTOR) group, 6 rats in each group. The first two groups of rats underwent 12 weeks of sham operation after ovariectomy, and the left maxillary second molars were ligatured to fabricate experimental models of periodontitis (ie, SHAM + hIL-10 + EP and SHAM + VECTOR + EP subgroups) The right side was not ligated as control teeth (ie SHAM + hIL-10 + C and SHAM + VECTOR + C subgroups were formed). The latter two groups of rats were performed 12 weeks after resection of bilateral ovarian surgery, the left maxillary second molars were ligatured and the model of experimental periodontitis was established (that is, the formation of OVX + hIL-10 + EP and OVX + VECTOR + EP subgroups) . At the same time, hIL-10 plasmid (5μg) was injected subcutaneouly into the palatal gingival mucosa of bilateral maxillary second molar in SHAM + hIL-10 group and the palatal palate side of the second maxillary second molar in OVX + hIL-10 group ) - liposome (5μL) complex; in the SHAM + VECTOR group rats bilateral maxillary second molar palatal gingival submucosa and OVX + VECTOR group rat left maxillary second molar palatal gingival submucosal injection Empty vector plasmid (5 μg) - liposome (5 μL) complex. One injection the next day, 48h after the seventh injection, rats were sacrificed. The bone mineral density, serum biochemical parameters, alveolar bone absorption and changes of periodontal tissue cytokines were observed. Results The number of IL-10 positive cells in the periodontal ligament of SHAM + hIL-10 + C group, SHAM + hIL-10 + EP group and OVX + hIL-10 + EP group were significantly decreased Higher than SHAM + VECTOR + C group, SHAM + VECTOR + EP group and OVX + VECTOR + EP group (all P <0.05). Compared with SHAM + VECTOR + C group, the number of IL-1β positive cells in SHAM + hIL-10 + C group was significantly decreased (P <0.05), and the number of RANKL positive cells was significantly increased (P <0.05). Compared with SHAM + VECTOR + EP group, the number of IL-1β, IL-6, TNF-α and RANKL positive cells in SHAM + hIL-10 + EP group were significantly decreased (P <0.05). Compared with OVX + VECTOR + EP group, the number of IL-1β, IL-6, RANKL and MMP-8 positive cells in the periodontal ligament of OVX + hIL-10 + EP group were significantly decreased (P <0.05). Conclusion Local injection of hIL-10 plasmid suppresses alveolar bone resorption in experimental periodontitis in ovariectomized rats, which may be related to the decrease of proinflammatory cytokine expression in periodontal tissues.