中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2009年
5期
270-273
,共4页
邓雨泉%付云%苏小鹏%唐志英
鄧雨泉%付雲%囌小鵬%唐誌英
산우천%부운%소소붕%당지영
细胞增殖%基质金属蛋白酶-1%细胞凋亡%晚期蛋白氧化产物
細胞增殖%基質金屬蛋白酶-1%細胞凋亡%晚期蛋白氧化產物
세포증식%기질금속단백매-1%세포조망%만기단백양화산물
Cell proliferation%Matrix metallopmteinase-1: Apoptosis%Advanced oxidative protein products
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(human gingival fibroblasts,HGF)增殖、凋亡及合成基质金属蛋白酶1(matrix matelloproteinast-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、100 mg/L AOPP-HSA的培养基,甲基嚷唑基四唑(MTT)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50 mg/L AOPP-HSA培养基共培养72 h,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、100 mg/L AOPP-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48 h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72 h后,0.5、5、50、100 mg/L AOPP.HAS组中MMP-1合成量[分别为(55.61±1.06)、(65.78±4.04)、(79.24±3.09)、(89.76±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.
目的 探討晚期蛋白氧化產物(advanced oxidative protein products,AOPP)對人牙齦成纖維細胞(human gingival fibroblasts,HGF)增殖、凋亡及閤成基質金屬蛋白酶1(matrix matelloproteinast-1,MMP-1)的影響,探討AOPP在糖尿病患者牙週病加重中的作用及其介導氧化應激的可能機製.方法 採用酶消化.組織塊培養法穫得HGF,在對照組中加入不含AOPP-人血清白蛋白(human serum albumin,HSA)的培養基,在實驗組中分彆加入含有5、50、100 mg/L AOPP-HSA的培養基,甲基嚷唑基四唑(MTT)比色法檢測在不同時間段下HGF增殖水平的變化;ELISA法測定HGF閤成MMP-1的水平;與含50 mg/L AOPP-HSA培養基共培養72 h,經膜聯蛋白V/PI雙染後,流式細胞儀檢測HGF的凋亡狀況.結果 5、50、100 mg/L AOPP-HSA組對HGF增殖抑製率與對照組比較差異均有統計學意義(P＜0.05),併在共培養的48 h達到峰值[分彆為(19.01±6.28)%、(30.48±5.75)%、(39.75±4.60)%],呈濃度依賴關繫;各實驗組HGF的凋亡率與對照組比較差異無統計學意義(P＞0.05);共培養72 h後,0.5、5、50、100 mg/L AOPP.HAS組中MMP-1閤成量[分彆為(55.61±1.06)、(65.78±4.04)、(79.24±3.09)、(89.76±28.88)mg/L]均高于對照組[(34.90±3.15)mg/L,P＜0.05],MMP-1水平與AOPP-HSA呈濃度依賴關繫.結論 AOPP能抑製成纖維細胞增值,該抑製作用不是通過誘導細胞凋亡來實現;AOPP可增加MMP-1的閤成,促進膠原溶解,加速牙週病進程.
목적 탐토만기단백양화산물(advanced oxidative protein products,AOPP)대인아간성섬유세포(human gingival fibroblasts,HGF)증식、조망급합성기질금속단백매1(matrix matelloproteinast-1,MMP-1)적영향,탐토AOPP재당뇨병환자아주병가중중적작용급기개도양화응격적가능궤제.방법 채용매소화.조직괴배양법획득HGF,재대조조중가입불함AOPP-인혈청백단백(human serum albumin,HSA)적배양기,재실험조중분별가입함유5、50、100 mg/L AOPP-HSA적배양기,갑기양서기사서(MTT)비색법검측재불동시간단하HGF증식수평적변화;ELISA법측정HGF합성MMP-1적수평;여함50 mg/L AOPP-HSA배양기공배양72 h,경막련단백V/PI쌍염후,류식세포의검측HGF적조망상황.결과 5、50、100 mg/L AOPP-HSA조대HGF증식억제솔여대조조비교차이균유통계학의의(P＜0.05),병재공배양적48 h체도봉치[분별위(19.01±6.28)%、(30.48±5.75)%、(39.75±4.60)%],정농도의뢰관계;각실험조HGF적조망솔여대조조비교차이무통계학의의(P＞0.05);공배양72 h후,0.5、5、50、100 mg/L AOPP.HAS조중MMP-1합성량[분별위(55.61±1.06)、(65.78±4.04)、(79.24±3.09)、(89.76±28.88)mg/L]균고우대조조[(34.90±3.15)mg/L,P＜0.05],MMP-1수평여AOPP-HSA정농도의뢰관계.결론 AOPP능억제성섬유세포증치,해억제작용불시통과유도세포조망래실현;AOPP가증가MMP-1적합성,촉진효원용해,가속아주병진정.
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 flbroblast(HGF).To explore the possible mechanism of the periodontal destruction acceleration in diabetes through AOPP-mediated oxidative stress.Methotis HGF were isolated bv 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/LAOPP-HAS group[(55.61±1.06),(65.78±4.04),(79.24±3.09),(89.76±28.88)mg/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 apeptosis.AOPP may increase collagen degradation by promoting MMP-1 synthesis and thus may accelerate periodontal destruction process in diabetes.
