【摘要】：目的探討白藜蘆醇(Resveratrol,Res)對(duì)基質(zhì)金屬蛋白酶-2(MMP-2)活性及耐脫礦力的影響,以期為白藜蘆醇發(fā)揮保護(hù)脫礦牙本質(zhì)基質(zhì)作用的臨床應(yīng)用提供實(shí)驗(yàn)依據(jù)。方法以體液基質(zhì)金屬蛋白酶-2活性比色法定量檢測(cè)試劑盒檢測(cè)白藜蘆醇對(duì)牙源性MMP-2抑制作用的有效濃度。利用激光共聚焦顯微鏡和掃描電鏡觀察經(jīng)pH循環(huán)處理后的牙本質(zhì)片脫礦程度及表面顯微形貌。以人I型膠原吡啶交聯(lián)終肽(ICTP)酶聯(lián)免疫分析各處理組牙本質(zhì)膠原纖維的降解情況。結(jié)果白藜蘆醇對(duì)基質(zhì)金屬蛋白酶-2活性的影響在一定濃度范圍內(nèi)呈現(xiàn)一定關(guān)系,即濃度在12.5-100umol/L時(shí),隨藥物濃度的增加,其對(duì)MMP-2的抑制作用明顯增加,但在100-400umol/L時(shí),其抑制作用較為穩(wěn)定。CLSM 3D圖像顯示牙本質(zhì)試件的脫礦深度:實(shí)驗(yàn)組100umol/LRes較陰性對(duì)照組小,與陽性對(duì)照組0.2%氯己定間無明顯差異。SEM觀察牙本質(zhì)試件顯微形貌:100umol/LRes處理組牙本質(zhì)表面脫礦程度較陰性對(duì)照組弱,表現(xiàn)為管周牙本質(zhì)脫礦不明顯,牙本質(zhì)小管管徑開放程度較陰性對(duì)照組小。且100umol/L Res組和0.2%氯己定組均可見于牙本質(zhì)小管口處現(xiàn)不同程度的顆粒沉積物。ICTP試劑盒結(jié)果表明:經(jīng)模擬人工齲損形成過程即pH循環(huán)處理后的牙本質(zhì)ICTP的釋放量,100umol/L Res組與0.2%氯己定組間無顯著性差異,與陰性對(duì)照組間有明顯差異,即100umol/L Res組和0.2%氯己定組ICTP釋放量顯著低于陰性對(duì)照組,表明陰性對(duì)照組的膠原纖維降解量明顯多于100umol/L Res組及0.2%氯己定組。結(jié)論白藜蘆醇可以抑制基質(zhì)金屬蛋白酶-2活性,從而抑制膠原降解及牙本質(zhì)脫礦,發(fā)揮保護(hù)脫礦牙本質(zhì)基質(zhì)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.
【學(xué)位授予單位】：佳木斯大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R781

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