CAJ | 학술논문

目的研究环孢素A(cyclosporinA)和肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)对体外培养人牙龈成纤维细胞增殖的影响,探讨牙龈炎症与药物性牙龈增生的关系及环孢素A所致牙龈增生的相关机制.方法用原代培养的方法获取5名健康人的牙龈成纤维细胞,体外培养、传代后取其中1个生长良好的组织块4～8代细胞用于实验.按以下条件进行实验分组:A组:空白对照组；B1组:10 μg/L环孢素A,B2组:50 μg/L环孢素A,B3组:250 μg/L环孢素A,B4组:1250 μg/L环孢素A；C组:5μg/L TNF-α; D1组:10 μg/L环孢素A+5μg/L TNF-α,D2组:50 μg/L 环孢素 A+5μg/L TNF-α,D3组:250 μg/L环孢素A+5μg/L TNF-α,D4组:1250 μg/L环孢素A+ 5 μg/L TNF-α.将条件培养液分别作用于牙龈成纤维细胞,培养3、5、7d后用甲基噻唑基四唑法测定细胞的增殖情况.结果不同质量浓度的环孢素A作用于成纤维细胞后,细胞的增殖受到抑制,A值下降,其中B1、B2、B3组与A组相比差异无统计学意义,B4组与A组相比差异具有统计学意义(P =0.001)；5μg/L的TNF-α作用于成纤维细胞可以刺激细胞的增殖,A值(0.542)与A组(0.441)相比显著升高(P＜0.01).环孢素A和TNF-α共同作用于成纤维细胞后,D1、D2、D3组A值均较A组升高,但均较C组显著降低(P＜0.05).D4组细胞增殖显著增加,与C组相比差异具有统计学意义(P＜0.01).结论环孢素A对成纤维细胞的增殖无促进作用,高浓度时可抑制细胞的增殖；TNF-α可以促进成纤维细胞的增殖；高浓度环孢素A与TNF-α共同作用于成纤维细胞可促进成纤维细胞增殖,提示在一定浓度下环孢素A可能放大了TNF-α刺激成纤维细胞增殖的效应.
목적 연구배포소A(cyclosporinA)화종류배사인자α(tumor necrosis factor-α,TNF-α)대체외배양인아간성섬유세포증식적영향,탐토아간염증여약물성아간증생적관계급배포소A소치아간증생적상관궤제.방법용원대배양적방법획취5명건강인적아간성섬유세포,체외배양、전대후취기중1개생장량호적조직괴4～8대세포용우실험.안이하조건진행실험분조:A조:공백대조조；B1조:10 μg/L배포소A,B2조:50 μg/L배포소A,B3조:250 μg/L배포소A,B4조:1250 μg/L배포소A；C조:5μg/L TNF-α; D1조:10 μg/L배포소A+5μg/L TNF-α,D2조:50 μg/L 배포소 A+5μg/L TNF-α,D3조:250 μg/L배포소A+5μg/L TNF-α,D4조:1250 μg/L배포소A+ 5 μg/L TNF-α.장조건배양액분별작용우아간성섬유세포,배양3、5、7d후용갑기새서기사서법측정세포적증식정황.결과불동질량농도적배포소A작용우성섬유세포후,세포적증식수도억제,A치하강,기중B1、B2、B3조여A조상비차이무통계학의의,B4조여A조상비차이구유통계학의의(P =0.001)；5μg/L적TNF-α작용우성섬유세포가이자격세포적증식,A치(0.542)여A조(0.441)상비현저승고(P＜0.01).배포소A화TNF-α공동작용우성섬유세포후,D1、D2、D3조A치균교A조승고,단균교C조현저강저(P＜0.05).D4조세포증식현저증가,여C조상비차이구유통계학의의(P＜0.01).결론배포소A대성섬유세포적증식무촉진작용,고농도시가억제세포적증식；TNF-α가이촉진성섬유세포적증식；고농도배포소A여TNF-α공동작용우성섬유세포가촉진성섬유세포증식,제시재일정농도하배포소A가능방대료TNF-α자격성섬유세포증식적효응.
Objective To investigate the effect of cyclosporin A(CsA) and tumor necrosis factor-α (TNF-α) on cell proliferation of cultured human gingival fibroblasts (GF),and the relationship between gingival inflammation and drug-induced gingival overgrowth.Methods Human GF were cultured in vitro using tissue culture method.then cells from the 4-8 th passage were used in the experiment.The cells were cultured and incubated with various concentrations of CsA and TNF-o (A:blank group,B1:10 μg/L CsA,B2:50 μg/L CsA,B3:250 μg/L CsA,B4:1250 μg/L CsA,C:5 μg/L TNF-α,D1:10 μg/L CsA + 5 μg/L TNF-α,D2:50 μg/L CsA +5 μg/L TNF-α,D3:250 μg/L CsA +5.μg/L TNF-α,D4:1250 μg/L CsA +5 μg/L TNF-α) solution for 3,5 and 7 days.Methyl thiazolyl tetrazolium assay was used to evaluate the cell proliferation in the culture meidiun.Results The proliferation of fibroblasts was inhibited when exposed to different concentration of CsA and A value decreased.There was no significant difference between group B1,B2,B3 and the control group,while the A value of group B4 was significantly higher than that of control group(P ＜ 0.01 ).Fibroblast proliferation was significantly increased while cultured with 5.μg/L TNF-α.A value increased( P ＜ 0.01 ).When exposed to CsA + TNF-α,A value of group D1,D2,D3 was much higher than that of group A,but was lower than that of group C ( P ＜ 0.05 ).Cell proliferation in group D4 was significantly increased,and significantly different with that in group C (P ＜ 0.01 ).Conclusions CsA did not stimulate the cell proliferation,and high concentration of CsA inhibited cell proliferation.TNF-α can stimulate the cell proliferation. High-concentration CsA + TNF-α can enhance the fibroblast proliferation,which suggests that CsA in certain concentration have amplification effect on TNF-α to stimulate fibroblast proliferation.