摘要
目的:观察柚皮素对支气管上皮细胞中Smad介导的转化生长因子β1(transforming growth factor β1,TGF-β1)通路的影响,并研究其可能的机制。方法:培养HBE细胞,分为对照组、TGF-β1组和柚皮素干预组。柚皮素干预组分别与25、50和100μmol/L的柚皮素共孵育2 h后,与TGF-β1组一起加入TGF-β1。2 h后用real-time PCR方法检测各组细胞纤溶酶原激活物抑制剂1(PAI-1)mRNA表达。另外在加入TGF-β1后30 min时,提取对照组、TGF-β1组和柚皮素高剂量组细胞总蛋白,用Western blot方法检测Smad2水平及磷酸化的Smad2水平,同时用免疫荧光法检测Smad2的核转位。结果:TGF-β1组的PAI-1mRNA水平较对照组明显增高,随着给予柚皮素剂量的增大,PAI-1 mRNA水平明显降低。TGF-β1组经TGF-β1刺激30 min后,磷酸化的Smad2明显增加,经柚皮素干预后则随着柚皮素剂量的增加,Smad2磷酸化明显受抑制。各组的总Smad2水平无明显变化。免疫荧光实验显示TGF-β1组的细胞核内Smad2明显增多,而柚皮素干预组(100μmol/L)的细胞核内Smad2含量明显减少。结论:柚皮素可能通过抑制Smad2磷酸化及减少核内Smad2含量来影响TGF-β1在支气管上皮细胞中的生物功能。
Objective：To investigate the effect of naringenin on TGF-β1/Smad pathway in human bronchial epithelial cells. Methods： HBE cells were divided into the control group,the TGF-β1 group and the naringenin group. The naringenin groups were incubated with different doses of naringenin （25,50 and 100 mol/L） for 2 h. Then the naringenin group and the TGF-β1 group were incubated with TGF-β1. After 2 h,we determined the PAI-I mRNA level by real time-PCR. After incubated with TGF-β1 for 30 min, we detected Smad2 and Smad2 phosphorylation by Western. Smad2 nuclear translocation was analyzed by indirect immunofluorescent staining. Results：Compared with the control group,the expression of PAI-1 mRNA was significantly increasing in HBE cells stimulated with TGF-β1. Naringenin has inhibitory effects on the expression of PAI-1 mRNA. The totle of Smad2 was similar in different groups. Compared with the control group,phosphorylated-Smad2 was increasing in the TGF-β1 group. Naringenin could significantly inhibit the Smad2 phosphorylation. Immunofluorescent staining showed that the nuclear content of Smad2 in the TGF-β1 group was increased,and in the naringenin group （100 μmol/L） was decreased. Conclusion： Naringenin may inhibit TGF-β1/Smad through attenuated Smad2 phosphorylation and suppressed the nuclear content of Smad2 in human bronchial epithelial cells.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2013年第2期155-159,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金项目(81100073)
