白藜芦醇对脱矿牙本质基质影响的研究

发布时间：2019-05-10 06:00

【摘要】：目的探讨白藜芦醇(Resveratrol,Res)对基质金属蛋白酶-2(MMP-2)活性及耐脱矿力的影响,以期为白藜芦醇发挥保护脱矿牙本质基质作用的临床应用提供实验依据。方法以体液基质金属蛋白酶-2活性比色法定量检测试剂盒检测白藜芦醇对牙源性MMP-2抑制作用的有效浓度。利用激光共聚焦显微镜和扫描电镜观察经pH循环处理后的牙本质片脱矿程度及表面显微形貌。以人I型胶原吡啶交联终肽(ICTP)酶联免疫分析各处理组牙本质胶原纤维的降解情况。结果白藜芦醇对基质金属蛋白酶-2活性的影响在一定浓度范围内呈现一定关系,即浓度在12.5-100umol/L时,随药物浓度的增加,其对MMP-2的抑制作用明显增加,但在100-400umol/L时,其抑制作用较为稳定。CLSM 3D图像显示牙本质试件的脱矿深度:实验组100umol/LRes较阴性对照组小,与阳性对照组0.2%氯己定间无明显差异。SEM观察牙本质试件显微形貌:100umol/LRes处理组牙本质表面脱矿程度较阴性对照组弱,表现为管周牙本质脱矿不明显,牙本质小管管径开放程度较阴性对照组小。且100umol/L Res组和0.2%氯己定组均可见于牙本质小管口处现不同程度的颗粒沉积物。ICTP试剂盒结果表明:经模拟人工龋损形成过程即pH循环处理后的牙本质ICTP的释放量,100umol/L Res组与0.2%氯己定组间无显著性差异,与阴性对照组间有明显差异,即100umol/L Res组和0.2%氯己定组ICTP释放量显著低于阴性对照组,表明阴性对照组的胶原纤维降解量明显多于100umol/L Res组及0.2%氯己定组。结论白藜芦醇可以抑制基质金属蛋白酶-2活性,从而抑制胶原降解及牙本质脱矿,发挥保护脱矿牙本质基质的作用。
[Abstract]:Objective to investigate the effect of resveratrol (Resveratrol,Res) on the activity and demineralization resistance of matrix metalloprotease-2 (MMP-2) in order to provide experimental basis for the clinical application of resveratrol in protecting demineralized dentin matrix. Methods the effective concentration of resveratrol on the inhibitory effect of resveratrol on odontogenic MMP-2 was detected by matrix metalloprotease-2 activity colorimetric assay. The demineralization degree and surface morphology of dentin treated with pH cycle were observed by laser confocal microscope and scanning electron microscope. The degradation of dentin collagen fibers in each treatment group was analyzed by enzyme-linked immunosorbent assay (Elisa) with human type I collagen pyridine crosslinked terminal peptide (ICTP). Results the effect of resveratrol on the activity of matrix metalloprotease-2 showed a certain relationship in a certain concentration range, that is, when the concentration of 12.5-100umol/L increased, the inhibitory effect of resveratrol on MMP-2 increased obviously with the increase of drug concentration. However, the inhibitory effect of 100-400umol/L was more stable. CLSM 3D images showed the demineralization depth of dentin specimens: the 100umol/LRes of the experimental group was smaller than that of the negative control group. There was no significant difference between the positive control group and the positive control group. The microstructure of dentin specimens was observed by SEM. the demineralization degree of dentin surface in 100umol/LRes treatment group was weaker than that in negative control group, which showed that peritubular dentin demineralization was not obvious. The opening degree of dentin tubule diameter was smaller than that of negative control group. In both 100umol/L Res group and 0.2% chlorhexidine group, granular deposits were observed in dentin tubule in varying degrees. The results of ICTP kit showed that the release of ICTP from dentin after simulated artificial caries formation process, that is, pH cycle treatment, was observed. There was no significant difference between 100umol/L Res group and 0.2% chlorhexidine group, but there was significant difference between 100umol/L Res group and negative control group, that is, ICTP release in 100umol/L Res group and 0.2% chlorhexidine group was significantly lower than that in negative control group. The results showed that the degradation of collagen fibers in negative control group was significantly higher than that in 100umol/L Res group and 0.2% chlorhexidine group. Conclusion resveratrol can inhibit the activity of matrix metalloprotease-2, inhibit collagen degradation and dentin demineralization, and play an important role in protecting demineralized dentin matrix.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781

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