摘要
目的 探讨多巴胺对基质金属蛋白酶活性及牙本质胶原纤维降解的影响,以期为多巴胺的临床应用提供实验依据.方法 以2.0 g/L多巴胺与1.0 g/LⅦ型胶原酶混合液为多巴胺组,以去离子水与1.0 g/LⅦ型胶原酶混合液为阴性对照组,以2％氯己定与1.0 g/LⅦ型胶原酶混合液为阳性对照组,每组成分混合体积比均为1∶9,每组重复5次,用细胞基质金属蛋白酶总活性比色法检测各组混合15 min后的酶活性.牙本质片用37％磷酸酸蚀1 min后,2个牙本质片直接观察表面形态,其余30个牙本质片根据随机数字表随机分为3组（每组10个）,阴性对照组：牙本质片放入100μl去离子水＋900μl 1.0 g/L胶原酶溶液中;阳性对照组：牙本质片放入100μl 2％氯己定＋900 μl 1.0 g/L胶原酶溶液中;多巴胺组：牙本质片放入100 μl2％多巴胺＋900μl 1.0 g/L胶原酶溶液中;3组均放入恒温孵箱（37℃）孵育7d后,检测孵化液中羟脯氨酸含量,计算胶原降解量,扫描电镜观察各组牙本质片胶原形态.结果 阴性对照组的酶活性及胶原降解量[（0.089±0.011） μmol＆#183;min-1＆#183;mg-1和（2 837±201） μg/cm2]均显著高于多巴胺组[（0.030±0.009）μmol＆#183;min-1＆#183;mg-1和（1 389±255） μg/cm2]和阳性对照组[（0.038±0.006）μmol＆#183;min-1＆#183;mg-1和（1 288±172） μg/cm2]（P＜0.05）,多巴胺组和阳性对照组间差异无统计学意义（P＞0.05）.扫描电镜显示多巴胺组和阳性对照组胶原纤维网结构完整,而阴性对照组胶原破坏,结构不完整.结论 多巴胺可抑制基质金属蛋白酶活性以及脱矿的牙本质胶原降解.
Objective To investigate the inhibition effect of dopamine on the activity of matrix metalloproteinases（MMP） and the effect of dopamine on degradation of dentin collagen for its potential use in caries treatment and dentin adhesive.Methods In the experiment of MMP activity test,2.0 g/L dopamine＋ 1.0 g/L highly purified collagenase type Ⅶ from Clostridium histolyticum served as the experimental group,and deionized water＋ 1.0 g/L highly purified collagenase type Ⅶ from Clostridium histolyticum served as the negative control group,and 2% chlorhexidine ＋ 1.0 g/L highly purified collagenase type Ⅶ from Clostridium histolyticum served as the positive control group,and the mixture volume ratio of the two ingredients in every group was 1∶9.After 15 minutes,the enzyme activity of each sample was tested by MMP activity colerimetric quantitative detection kits,and the test was repeated 5 times in each group.In the experiment of collagen degradation,the dentin slices were demineralized with 37% phosphoric acid for 1 min.In sequence,2 dentin slices were used to observe the morphology,and the remaining 30 dentine slices were randomly divided into three groups（n=10） according to random number table：the negative control ones were stored in 100 μl deionized water and 900 μl collagenase（7 days,37 ℃）,the positive control ones were stored in 100 μl chlorhexidine and 900 μl collagenase（7 days,37 ℃） and the experimental specimens were stored in 100 μl dopamine and 900 μl collagenase（7 days,37 ℃）.The degraded collagen was investigated by assaying hydroxyproline.The framework of collagen was evaluated with field emission scanning electron microscope（FE-SEM）.Results The statistical results of completely random design ANOVA showed that the MMP activity and the amount of degraded collagen of the negative control group [（0.089±0.011） μmol ＆#183; min-1＆#183; mg-1 and （2 837±201） μg/cm2] were significantly higher than those of the positive control group[（0.038± 0.006） μmol ＆#183; min-1 ＆#183; mg-1 and （1 288± 172） μg/cm2] and the experimental group[（0.030±0.009） μmol＆#183; min-1＆#183; mg-1 and （1 389±255） μg/cm2]（P〈0.05）.SEM observation indicated that the structural integrity of the collagen network on dentin still existed in experiment samples and positive control groups,however,collagen fibrils were destructed and the structural integrity disappeared in the negative control groups.Conclusions Dopamine may inhibit MMP activity and reduce the amount of degraded collagen.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2015年第3期186-189,共4页
Chinese Journal of Stomatology
基金
国家自然科学基金（81400559）
国家自然科学基金对外交流与合作项目（81061160511）
