大鼠正畸过程中牙周组织糖酵解途径变化

Changes of glycolytic pathway during orthodontic treatment in rats

目的探讨大鼠正畸过程中牙周组织糖酵解途径变化。方法48只雄性SD大鼠按照随机数字表法随机分为对照组、正畸7 d组、正畸14 d组,建立大鼠正畸牙齿移动模型。采用游标卡尺测量大鼠正畸治疗后牙齿移动距离;采用酶联免疫吸附法(ELISA)测定大鼠牙槽骨组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6表达水平;采用微量法检测大鼠牙槽骨组织中糖酵解途径关键酶丙酮酸激酶(PK)、乳酸脱氢酶(LDH)、磷酸果糖激酶(PFK)的活性;采用蛋白质印迹法(Western blot)检测糖酵解上游调控因子信号转导及转录激活蛋白3(STAT3)、缺氧诱导因子-1α(HIF-1α)的蛋白表达水平,计量数据比较采用t检验。结果正畸7 d组[(0.476±0.017)mm]、正畸14 d组[(0.715±0.027)mm]大鼠牙齿移动距离明显高于对照组(0 mm),差异有统计学意义(t=16.563、20.730,P<0.05)。正畸7 d组[(139.411±5.985)ng/ml]、正畸14 d组[(196.068±8.972)ng/ml]大鼠牙槽骨组织中TNF-α的水平明显高于对照组[(33.556±2.756)ng/ml],差异有统计学意义(t=10.361、13.426,P<0.05)。正畸7 d组[(297.829±10.309)ng/ml]、正畸14 d组[(372.771±14.675)ng/ml]IL-1β的水平明显高于对照组[(127.656±4.274)ng/ml],差异有统计学意义(t=7.852、10.446,P<0.05)。正畸7 d组[(251.620±10.925)ng/ml]、正畸14 d组[(320.611±14.981)ng/ml]IL-6的水平明显高于对照组[(94.633±4.394)ng/ml],差异有统计学意义(t=9.156、12.749,P<0.05)。正畸7 d组[(109.654±5.093)U/mgprot]、正畸14 d组[(163.626±5.282)U/mg蛋白]大鼠牙槽骨组织中PK的活性明显高于对照组[(57.083±2.852)U/mg蛋白],差异有统计学意义(t=6.363、9.532,P<0.05)。正畸7 d组[(120.173±7.187)U/mg蛋白]、正畸14 d组[(171.069±6.230)U/mg蛋白]LDH的水平明显高于对照组[(78.421±4.030)U/mg蛋白],差异有统计学意义(t=5.580、7.644,P<0.05)。正畸7 d组[(38.503±1.895)U/mg蛋白]、正畸14 d组[(51.411±3.095)U/mg蛋白]PFK的水平明显高于对照组[(24.236±1.200)U/mg蛋白],差异有统计学意义(t=5.591、7.447,P<0.05)。正畸7 d组(1.099±0.050)、正畸14 d组(1.494±0.053)大鼠牙槽骨组织中STAT3的蛋白表达水平明显高于对照组(0.604±0.045),差异有统计学意义(t=7.066、9.332,P<0.05)。正畸7 d组(1.317±0.049)、正畸14 d组(1.664±0.058)HIF-1α的蛋白表达水平明显高于对照组(0.748±0.043),差异有统计学意义(t=6.248、8.693,P<0.05)。结论大鼠正畸牙齿移动过程中牙周组织糖酵解水平明显增强,并可能进一步加剧正畸过程中牙周组织的炎性反应。

Objective To investigate the changes of glycolytic pathway in periodontal tissue and the clinical significance during orthodontic treatment in rats.Methods Totally,48 male SD rats were randomly divided into control group,orthodontic 7 d group and orthodontic 14 d group by a random number table.The orthodontic tooth movement model was established.The tooth movement distance was measured by vernier calipers after orthodontic treatment.The expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 in alveolar bone tissue of rats were determined by enzyme linked immunosorbent assay(ELISA).The activities of pyruvate kinase(PK),lactate dehydrogenase(LDH)and phosphofructokinase(PFK)in the alveolar bone of rats were detected by microassay.The Western blotting was used to detect the protein expression of signal transducer and activator of transcription 3(STAT3)and hypoxiaduciblefactors-1α(HIF-1α),upstream regulators of glycolysis.The measurement data were compared by t test.Results Compared with the control group(0 mm),the tooth movement distance of the rats in the orthodontic 7 d group[(0.476±0.017)mm]and the orthodontic 14 d group[(0.715±0.0.027)mm]was significantly increased(t=16.563,20.730,P<0.05).The expression of TNF-α in the orthodontic 7 d group[(139.411±5.985)ng/ml]and orthodontic 14 d group[(196.068±8.972)ng/ml]was significantly higher than that in the control group[(33.556±2.756)ng/ml,t=10.361,13.426,P<0.05].The expression of IL-1β in the orthodontic 7 d group[(297.829±10.309)ng/ml]and orthodontic 14 d group[(372.771±14.675)ng/ml]was significantly higher than in the control group[(127.656±4.274)ng/ml,t=7.852,10.446,P<0.05].The expression levels of IL-6 in orthodontic 7 d group[(251.620±10.925)ng/ml]and orthodontic 14 d group[(320.611±14.981)ng/ml]were significantly higher than in the control group[(94.633±4.394)ng/ml,t=9.156,12.749,P<0.05].The PK activity in the orthodontic 7 d group[(109.654±5.093)U/mg prot]and the orthodontic 14 d group[(163.626±5.282)U/mg prot,t=6.363,9.532,P<0.05].The LDH activity in the orthodontic 7 d group[(120.173±7.187)U/mg prot]and orthodontic 14 d group[(171.069±6.230)U/mg prot]was significantly higher than in the control group[(78.421±4.030)ng/ml????,t=5.580,7.644,P<0.05].The PFK activity in orthodontic 7 d group[(38.503±1.895)U/mg prot]and orthodontic 14 d group[(51.411±3.095)U/mg prot]was significantly higher than that in the control group[(24.236±1.200)ng/ml,t=5.591,7.447,P<0.05].The protein expression of STAT3 in the orthodontic 7 d group(1.099±0.050)and the orthodontic 14 d group(1.494±0.053)was significantly higher than that in the control group(0.604±0.045,t=7.066,9.332,P<0.05).The protein expression of HIF-1α in the orthodontic 7 d group(1.317±0.049)and orthodontic 14 d group(1.664±0.058)was significantly increased as compared with that in the control group(0.748±0.043,t=6.248,8.693,P<0.05).Conclusion The glycolysis level of periodontal tissue was significantly increased in rats during orthodontic tooth movement,which may further aggravate the inflammatory response of periodontal tissue.