晚期蛋白氧化产物对人牙龈成纤维细胞增殖、凋亡及合成基质金属蛋白酶1的影响  

Biologic effects of advanced oxidative protein products on the human gingival fibroblasts

作　　者：邓雨泉[1] 付云[1] 苏小鹏[1] 唐志英[1] 

机构地区：[1]中山大学光华口腔医学院牙周科,广州510055

出　　处：《中华口腔医学杂志》2009年第5期270-273,共4页Chinese Journal of Stomatology

摘　　要：目的探讨晚期蛋白氧化产物（advanced oxidative protein products，AOPP）对人牙龈成纤维细胞（humangin gival fibroblasts，HGF）增殖、凋亡及合成基质金属蛋白酶1（matrixmatelloproteinast-1，MMP-1）的影响，探讨AOPP在糖尿病患者牙周病加重中的作用及其介导氧化应激的可能机制。方法采用酶消化．组织块培养法获得HGF，在对照组中加入不含AOPP-人血清白蛋白（human serum albumin，HSA）的培养基，在实验组中分别加入含有5、50、100mg／LAOPP—HSA的培养基，甲基噻唑基四唑（Mrrr）比色法检测在不同时间段下HGF增殖水平的变化；ELISA法测定HGF合成MMP-1的水平；与含50mg／LAOPP—HSA培养基共培养72h，经膜联蛋白V／PI双染后，流式细胞仪检测HGF的凋亡状况。结果5、50、100mg／LAOPP—HSA组对HGF增殖抑制率与对照组比较差异均有统计学意义（P〈0．05），并在共培养的48h达到峰值[分别为（19．01±6．28）％、（30．48±5．75）％、（39．75±4．60）％]，呈浓度依赖关系；各实验组HGF的凋亡率与对照组比较差异无统计学意义（P〉0．05）；共培养72h后，0．5、5、50、100mg／LAOPP—HAS组中MMP-1合成量[分别为（55．61±1．06）、（65．78±4．04）、（79．24±3．09）、（89．764-28．88）mg／L]均高于对照组[（34．90±3．15）mg／L，P〈0．05]，MMP-1水平与AOPP—HSA呈浓度依赖关系。结论AOPP能抑制成纤维细胞增值，该抑制作用不是通过诱导细胞凋亡来实现；AOPP可增加MMP-1的合成，促进胶原溶解，加速牙周病进程。Objective To investigate the effects of advanced oxidative protein products （AOPP） on the proliferation, apoptosis and matrix matelloproteinase-1 （MMP-1） synthesis of the human gingival fibroblast（HGF）. To explore the possible mechanism of the periodontal destruction acceleration in diabetes through AOPP-mediated oxidative stress. Methods HGF were isolated by both tissue explant cultivation technique and enzyme digestion method. The culture media with 5,50,100 mg/L AOPP-HAS were added into each experimental group, but the culture media in the control group didn＇t contain AOPP-HAS. MTT colorimetric assay and ELISA were used to measure the changes of HGF proliferation and the levels of MMP-1 protein from HGF at different time periods, respectively. Seventy-two hours after co-culture with 50 mg/L AOPP-HSA, cell apoptosis was detected by flow cytometry with Annexin V/PI staining. Results Compared to the control group, the growth inhibition rate of HGF in 5,50,100 mg/L AOPP-HSA group was significantly different（P 〈 0. 05）. The peak value appeared at 48 hours of co-culture [ （ 19. 01 ± 6. 28 ）%, （30. 48 ±5.75 ） % , （39.75±4. 60） % , respectively] . There was a dose-dependent relationship between the growth inhibition rate and AOPP-HSA. No significant difference was detected on the apoptotic level between experimental group and the control （P 〉0. 05）. The MMP-1 synthesis in 0.5,5 ,50,100 mg/L AOPP-HASgroup [（55.61 ±1.06）, （65.78±4.04）, （79.24 ± 3.09）, （89.76 ±28.88） rag/L, respectively] was significantly higher than that in the control [ （34. 90 ± 3.15） mg/L] after 72 hours coculture （ P 〈 0. 05 ）. There was a dose-dependent relationship between MMP-1 and AOPP-HSA. Conclusions AOPP may inhibit the proliferation of HGF and such effect was not achieved through apoptosis. AOPP may increase collagen degradation by promoting MMP-1 synthesis and thus may accelerate periodontal destruction process in diabetes.

关 键 词：细胞增殖 基质金属蛋白酶-1 细胞凋亡 晚期蛋白氧化产物 

分 类 号：R686[医药卫生—骨科学]