牙周治疗对伴动脉粥样硬化牙周炎白细胞介素6及基质金属蛋白酶影响的动物实验

Influence of periodontal therapy on serum interleukin-6 and carotid metalloproteinases in animals with chronic periodontitis associated with atherosclerosis

目的 通过研究牙周基础治疗对伴动脉粥样硬化（atherosclerosis,As）慢性牙周炎（chronic periodontitis,CP）大鼠的影响,以期为临床提供参考.方法 44只清洁级SD大鼠按随机数字表法随机分为A组（正常对照组,7只）、B组（As组,建立As模型,8只）及C组（CP＋ As组,建立CP＋ As模型,29只）.建模后将C组再应用随机数字表法分为4个小组,CI组：自然进程组,C2组：单纯机械治疗组,C3组：机械治疗＋盐酸米诺环素（派丽奥）组,C4组：机械治疗＋盐酸米诺环素＋抗生素组,每组7只.建模后15周按随机数字表法处死B、C组大鼠各1只,分别进行血管病理切片发现有脂质沉积形成,同时观察C组大鼠口腔出现牙周炎表现,证明As及CP＋ As建模成功,开始口腔干预.分别于建模后第1、3、5、7和9周应用酶联免疫吸附测定法检测血清中白细胞介素6（interleukin-6,IL-6）的浓度,免疫组化法定性检测各组大鼠颈动脉血管壁组织的基质金属蛋白酶（matrix metalloproteinase,MMP）2、MMP-9并测量其灰度值.结果 各组MMP-2,9的灰度值趋势基本一致,B、C组颈动脉的内膜、中膜阳性染色深度均明显大于A组.C1组（MMP-2,9的灰度值分别为101.3±2.4、101.2±4.1）有As斑块形成,染色深度明显强于B组（MMP-2,9的灰度值分别为12.6.4±2.0、124.8±2.8）（P＜0.05）;C2、C3和C4组染色范围及程度依次减弱,且均弱于C1组,与C1组的灰度值相比差异均有统计学意义（P＜0.05）.血清检测结果显示,随着时间推移,B、C1组血清IL-6浓度一直呈升高趋势,均显著高于A组（P＜0.01）;C2、C3、C4组血清IL-6浓度均为先升后降,在建模后第5周达峰值,其中C2组最高,从第4次取样时间点起,各组IL-6含量均显著低于C1组（P＜0.001）,C3、C4组显著低于C2组（P＜0.01）.结论 对于伴有As的CP大鼠,若不及时控制CP,可能会加重As的病变程度;牙周机械治疗可能会短期内增高机体炎症反应程度,而长期则可能降低血管炎症,减缓其As病变程度;牙周机械治疗时辅以局部及全身抗炎药物后,颈动脉血管病变及全身炎症可以得到更好的改善.

Objective To investigate the effect of periodontal mechanical treatment on serum interleukin-6 （IL-6） and carotid artery matrix metalloproteinase（MMP）-2 and MMP-9 expression in chronic periodontitis（CP） SD rats with atherosclerosis （As）.Methods Forty-four six-week-old male SD rats were randomly divided into three groups：control group（group A）,As group （group B）,As ＋ CP group （group C）.According to different periodontal interventions,group C was randomly subdivided into four groups：natural process group （C 1）,the periodontal mechanical treatment group （C2）,the periodontal mechanical treatment ＋ local drugs group（C3）,and the periodontal mechanical treatment ＋ local and system drugs group（C4）.Each group received the appropriate treatment and periodontal interventions.Serum IL-6 levels were determined by enzyme linked immunosorrbent assay （ELISA）.MMP-2 and MMP-9 levels in the proximal aorta were examined by immunohistochemistry.Results The gray value of MMP-2 and MMP-9 was basically the same in all groups.Compared with group A,the gray value of MMP-2 and MMP-9 of group B and C were decreased.C1 group showed the formation of atherosclerotic plaque and fibrous cap.Compared with group B （126.4 ± 2.0,124.8 ± 2.8）,the gray value of group C1 （101.3 ± 2.4,101.2 ± 4.1） was significantly weaker（P ＜ 0.05）.The staining depth of MMP-2 and MMP-9 of groups C1,C2,C3 and C4 were sequentially decreased,and the differences of gray value were statistically significant（P ＜ 0.05）.The levels of serum IL-6 in groups B and C1 increased gradually with time and became significantly higher than that of group A（P ＜ 0.01）.The levels of serum IL-6 in groups C2,C3,and C4 increased gradually and reached the peak 5 weeks after the establishment of model （P ＜ 0.001）.After that,the levels of serum IL-6 decreased gradually and was lower than baseline.The levels of serum IL-6 in groups C3 and C4 were significantly lower than that in group C2 7 weeks after the establishment of model（P ＜ 0.01）.Conclusions In rats with periodontitis and cardiovascular diseases,chronic periodontal inflammation may significantly increase the severity of As and promote the formation of atherosclerotic plaque.Mechanical periodontal therapy may cause short-term systemic inflammation and then reduce vascular inflammation in long term.With supplement use of local and systemic antibiotics,the mechanical periodontal therapy may get the vascular disease and systemic inflammation improved.