摘要
目的 探讨红景天苷(Sal)调控Notch1/Hes1通路对人瘢痕疙瘩成纤维细胞(HKF)增殖、迁移和胶原合成的影响.方法 取对数生长期的HKF,分为对照组(NC组),Sal-L、Sal-M、Sal-H(10、20、40 μmol·L-1)组,Jagged 1(Notch 激活剂,5 mg·L-1)组,Sal(40 μmol·L-1)+Jagged1(5 mg·L-1)组.MTT 法检测细胞增殖能力,流式细胞术检测细胞周期和细胞凋亡,划痕实验检测细胞迁移,Western blot法检测细胞中胶原蛋白(collagen)-Ⅰ、collagen-Ⅲ、增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、Notch 1、Hes1蛋白表达.结果 与NC组比较,Sal-L、Sal-M、Sal-H组细胞A值及S期、G2/M期细胞比例下降,划痕愈合率减小,collagen-Ⅰ、collagen-Ⅲ、PCNA、MMP-2、Notch 1、Hes1蛋白表达降低,G0/G1期细胞比例、细胞凋亡率升高,且呈浓度依赖性(均P＜0.05),而Jagged 1组上述指标改变均相反.与Sal-H组比较,Sal+Jagged1组细胞A值、S期和G2/M期细胞比例、划痕愈合率增加,collagen-Ⅰ、collagen-Ⅲ、PCNA、MMP-2、Notch 1、Hes1蛋白表达升高,G0/G1期细胞比例、细胞凋亡率降低(P＜0.05).结论 Sal可能通过抑制Notch1/Hes1通路抑制HKF增殖、迁移及胶原合成.
Abstract
AIM To investigate the effects of salidroside(Sal)on the proliferation,migration and collagen synthesis of human keloid fibroblasts(HKF)by regulating Notch 1/Hes1 pathway.METHODS HKF in logarithmic growth period was divided into control group(NC group),Sal-L,Sal-M,Sal-H(10,20,40 μmol·L-1)group,Jagged1 group(Notch activator,5 mg·L-1),Sal(40 μmol·L-1)+Jagged1(5 mg·L-1)group.MTT assay was used to detect cell proliferation.Cell cycle and apoptosis were detected by flow cytometry.Cell migration was detected by scratch test,and Western blot was used to detect the expression of collagen-Ⅰ,collagen-Ⅲ,proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),Notch1 and Hes1 proteins.RESULTS Compared with the NC group,the A value,the proportion of cells in S phase and G2/M phase decreased,scratch healing rate decreased,the protein expression of collagen-Ⅰ,collagen-Ⅲ,PCNA,MMP-2,Notch1 and Hes1 decreased in the Sal-L,Sal-M and Sal-H group,the proportion of cells in G0/G1 phase and the rate of apoptosis increased,in a dose-dependent manner(P＜0.05),while the Jagged1 group showed the opposite change of the above indexes.Compared with the Sal-H group,the A value,the proportion of cells in S phase and G2/M phase,scratch healing rate,the protein expression of collagen-Ⅰ,collagen-Ⅲ,PCNA,MMP-2,Notch1 and Hes1 increased,the proportion of cells in G0/G1 phase and the rate of apoptosis decreased in the Sal group(P＜0.05).CONCLUSION Sal may inhibit the proliferation,migration and collagen synthesis of HKF by inhibiting Notch1/Hes1 pathway.
维普
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