CAJ | 학술논문

目的拟观察5-氟尿嘧啶对瘢痕疙瘩成纤维细胞Smad7和β转化生长因子(transforming growth factor-6,TGF-β)Ⅰ型受体以及细胞凋亡相关基因Bcl-2和Bax蛋白表达的影响.方法 (1)瘢痕疙瘩组织成纤维细胞原代培养,取4～6代传代细胞加入5个不同浓度(10,20,40,80,160μmol/L)5-氟尿嘧啶干预24,48,72 h.(2)利用四甲基偶氮唑盐法(MTT)测定瘢痕疙瘩成纤维细胞的增殖能力.(3)应用蛋白免疫印迹法(Western-blot)检测各组搬痕疙瘩成纤维细胞中Smad7和TGF-βⅠ型受体的表达及对Bcl-2、Bax蛋白表达的影响.结果 (1)MTT实验中,当5-氟尿嘧啶浓度为10,20 μmol/L作用24 h时,未发现成纤维细胞死亡,与空白对照组相比差异无统计学意义(P＞0.05);其他浓度下作用各组成纤维细胞死亡现象差异均有统计学意义(P＜0.01).(2)免疫印迹法实验结果如下:①与空白对照组相比,TGF-β1组Smad7表达明显减弱,TGF-βⅠ型受体表达则显著增强,差异有统计学意义(P＜0.01).②加入5-氟尿嘧啶干预后可显著增强Smad7的表达,差异有统计学意义(P＜0.01).③经不同浓度5-氟尿嘧啶干预后,实验组Bcl-2蛋白表达均低于对照组,差异有统计学意义(P＜0.05).但实验组Bax蛋白表达均高于对照组(P＜0.05),TGF-βⅠ型受体表达无明显影响,差异均无统计学意义(P＞0.05).结论 5-氟尿嘧啶对瘢痕疙瘩成纤维细胞具有抑制增殖和诱导凋亡的作用.
목적 의관찰5-불뇨밀정대반흔흘탑성섬유세포Smad7화β전화생장인자(transforming growth factor-6,TGF-β)Ⅰ형수체이급세포조망상관기인Bcl-2화Bax단백표체적영향.방법 (1)반흔흘탑조직성섬유세포원대배양,취4～6대전대세포가입5개불동농도(10,20,40,80,160μmol/L)5-불뇨밀정간예24,48,72 h.(2)이용사갑기우담서염법(MTT)측정반흔흘탑성섬유세포적증식능력.(3)응용단백면역인적법(Western-blot)검측각조반흔흘탑성섬유세포중Smad7화TGF-βⅠ형수체적표체급대Bcl-2、Bax단백표체적영향.결과 (1)MTT실험중,당5-불뇨밀정농도위10,20 μmol/L작용24 h시,미발현성섬유세포사망,여공백대조조상비차이무통계학의의(P＞0.05);기타농도하작용각조성섬유세포사망현상차이균유통계학의의(P＜0.01).(2)면역인적법실험결과여하:①여공백대조조상비,TGF-β1조Smad7표체명현감약,TGF-βⅠ형수체표체칙현저증강,차이유통계학의의(P＜0.01).②가입5-불뇨밀정간예후가현저증강Smad7적표체,차이유통계학의의(P＜0.01).③경불동농도5-불뇨밀정간예후,실험조Bcl-2단백표체균저우대조조,차이유통계학의의(P＜0.05).단실험조Bax단백표체균고우대조조(P＜0.05),TGF-βⅠ형수체표체무명현영향,차이균무통계학의의(P＞0.05).결론 5-불뇨밀정대반흔흘탑성섬유세포구유억제증식화유도조망적작용.
Objective To investigate the of 5-fluorouracil effects on the expression of Smad7,TGF-β receptorⅠ,Bcl-2 and Bax in keloid fibroblasts.Methods After primary culture of keloid fibroblasts,4-6 passages of cells were inoculated in 5 different concentrations of 5-fluorouracil(10,20,40,80,160μmol/L)for 24,48 and 72 hours.Proliferative ability of keloid fibroblasts was detected by MTT assay.Expression of Smad 7,TGF-βreceptorⅠ,Bcl-2 and Bax in keloid fibroblasts was measured by Western blot.Results During MTT,5-fluorouracil did not affect cell viability at 24 hour at the concentration of 10 and 20 μmol/L.Compared with the control group,no significant difference was detected(P＞0.05).At other concentrations,fibroblast death was visible in each group(P＜0.01).Western blot analysis showed that the expression of Smad7 significantly decreased and the expression of TGF-β receptor Ⅰ significantly increased in the TGF-β1 group compared with the blank control group(P＜0.0 1).5-fluorouracil could significantly enhance the expression of Smad7(P＜0.01).There was a remarkable decrease of the Bcl-2 expression and marked increase of the Bax expression in different concentrations of 5-fluorouracil compared with the control group(P＜0.05).But,5-fluorouracil did not show any effect on the synthesis of TGF-β receptor Ⅰ.Conclusion 5-fluorouracil could inhibit proliferation and induce apoptosis on human keloid fibroblasts in vitro.