中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2008年
1期
12-15
,共4页
糖基化终产物,高级%成纤维细胞%胶原%细胞增殖
糖基化終產物,高級%成纖維細胞%膠原%細胞增殖
당기화종산물,고급%성섬유세포%효원%세포증식
Glycosylation end products,advanced%Fibroblasts%Collagen%Cell proliferation
目的 观察糖基化终产物对牙龈成纤维细胞增殖能力和Ⅰ型胶原合成的影响,探讨糖基化终产物对牙周组织修复功能的作用及在伴有糖尿病牙周炎中的致病机制.方法 采用组织块法培养牙龈成纤维细胞,培养基中加入体外合成的不同浓度的糖基化终产物,甲基噻唑基四唑(MTT)法检测在不同时间段下牙龈成纤维细胞增殖水平的变化,酶联免疫吸附测定法和实时定量反转录聚合酶链法(RT-PCR)分别测定牙龈成纤维细胞合成Ⅰ型胶原的量和表达Ⅰ型胶原mRNA的水平.结果 200 mg/L糖基化终产物作用于牙龈成纤维细胞48、74 h的A值分别为0.016±0.023、0.035±0.008,比其他组A值低(P＜0.05),并且细胞形态发生了改变,细胞由均一的梭形变成圆形、椭圆形、细长不规则条状等,胞质浓缩,细胞间隙增大;糖基化终产物作用72 h后,牙龈成纤维细胞上清液中和胞内Ⅰ型胶原的含量减少(P＜0.05),Ⅰ型胶原mRNA表达下调(P＜0.05).结论 糖基化终产物能抑制牙龈成纤维细胞的增殖、降低其合成Ⅰ型胶原蛋白和表达Ⅰ型胶原mRNA,从而可能削弱了牙周组织的愈合能力.
目的 觀察糖基化終產物對牙齦成纖維細胞增殖能力和Ⅰ型膠原閤成的影響,探討糖基化終產物對牙週組織脩複功能的作用及在伴有糖尿病牙週炎中的緻病機製.方法 採用組織塊法培養牙齦成纖維細胞,培養基中加入體外閤成的不同濃度的糖基化終產物,甲基噻唑基四唑(MTT)法檢測在不同時間段下牙齦成纖維細胞增殖水平的變化,酶聯免疫吸附測定法和實時定量反轉錄聚閤酶鏈法(RT-PCR)分彆測定牙齦成纖維細胞閤成Ⅰ型膠原的量和錶達Ⅰ型膠原mRNA的水平.結果 200 mg/L糖基化終產物作用于牙齦成纖維細胞48、74 h的A值分彆為0.016±0.023、0.035±0.008,比其他組A值低(P＜0.05),併且細胞形態髮生瞭改變,細胞由均一的梭形變成圓形、橢圓形、細長不規則條狀等,胞質濃縮,細胞間隙增大;糖基化終產物作用72 h後,牙齦成纖維細胞上清液中和胞內Ⅰ型膠原的含量減少(P＜0.05),Ⅰ型膠原mRNA錶達下調(P＜0.05).結論 糖基化終產物能抑製牙齦成纖維細胞的增殖、降低其閤成Ⅰ型膠原蛋白和錶達Ⅰ型膠原mRNA,從而可能削弱瞭牙週組織的愈閤能力.
목적 관찰당기화종산물대아간성섬유세포증식능력화Ⅰ형효원합성적영향,탐토당기화종산물대아주조직수복공능적작용급재반유당뇨병아주염중적치병궤제.방법 채용조직괴법배양아간성섬유세포,배양기중가입체외합성적불동농도적당기화종산물,갑기새서기사서(MTT)법검측재불동시간단하아간성섬유세포증식수평적변화,매련면역흡부측정법화실시정량반전록취합매련법(RT-PCR)분별측정아간성섬유세포합성Ⅰ형효원적량화표체Ⅰ형효원mRNA적수평.결과 200 mg/L당기화종산물작용우아간성섬유세포48、74 h적A치분별위0.016±0.023、0.035±0.008,비기타조A치저(P＜0.05),병차세포형태발생료개변,세포유균일적사형변성원형、타원형、세장불규칙조상등,포질농축,세포간극증대;당기화종산물작용72 h후,아간성섬유세포상청액중화포내Ⅰ형효원적함량감소(P＜0.05),Ⅰ형효원mRNA표체하조(P＜0.05).결론 당기화종산물능억제아간성섬유세포적증식、강저기합성Ⅰ형효원단백화표체Ⅰ형효원mRNA,종이가능삭약료아주조직적유합능력.
Objective To apply the synthesized advanced glycation end products(AGE)to the cultured human gingival fibroblast(HGF)in vitro and then to investigate the effects of AGE on the HGF proliferation and type Ⅰ collagen synthesis and the potential impact of AGE in the repair of periodontium and its molecular mechanism in diabetes-associated periodontitis. Methods The HGF was obtained from explants of human healthy gingival tissues by using tissue-explant technique.The AGE Was prepared and then added to the culture media, its effect on HGF proliferation at different time duration Was examined with MTT colorimetric assay.The type Ⅰ collagen concentrations in cell culture supematants and intracellular proteins were detected by EUSA,and the type Ⅰ collagen mRNA expression of HGF was analyzed by realtime RT-PCR. Results 200 mg/L AGE decreased the A value(P ＜0.05)and changed the HGF shape.Incubation of HGF with AGE for 72 hours,the quantities of type Ⅰ collagen were reduced(P＜0.05),and the expression of type Ⅰ collagen mRNA Was down-regulated(P＜0.05). Condusions The AGE inhibited the HGF proliferation,decreased the synthesis of type Ⅰ collagen and down-regulated the expression of type Ⅰ collagen mRNA,impairing the repair of periodontium.
