摘要
目的研究苦参碱对肺成纤维细胞转化生长因子-β1(TGF-β1)信号转导途径的影响。方法以人肺成纤维细胞系(HLF-02)为研究对象,用Western blot法检测TGF-β1、丝/苏氨酸激酶抑制剂星形孢菌素(Staurosporine,SP)和细胞外信号调节激酶(ERK1/2)抑制剂PD98059、苦参碱(Matrine,Mat)作用对结缔组织生长因子(CTGF)、p-Smad2、p-ERK1/2蛋白表达的影响,免疫组织化学方法检测CTGF蛋白,RT-PCR技术检测CTGFmRNA的表达。结果体外培养的HLF-02表达基础水平的CTGF蛋白,1 ng/mL TGF-β1可使CTGF、CTGFmRNA、p-Smad2、p-ERK1/2表达水平增强(P<0.01);使用SP及PD98059预处理细胞可以抑制TGF-β1刺激的CTGF的表达增加(P<0.01)。使用苦参碱预处理细胞可以抑制TGF-β1刺激的CTGF、CTGFmRNA、p-Smad2、p-ERK1/2蛋白的表达(P<0.05或P<0.01),且与苦参碱浓度呈负相关(r分别为0.845,0.900,0.789,0.942;P<0.01)。结论TGF-β1可使CTGF蛋白及CTGFmRNA的表达明显增强,可能是通过Smad2和ERK途径促进CTGF表达。苦参碱可能是通过Smad2和ERK通路在基因及蛋白水平抑制TGF-β1诱导的CTGF表达。
Objective To investigate the effect of matrine on the signal transduction pathway of transforming growth factor-β 1（TGF-β 1）.Methods The effect of TGF-β1,Staurosporine,PD98059 and matrine on the expression of CTGF,CTGF mRNA,p-Smad2,and p-ERK 1/2 were evaluated by Western blot in cultured HLF-02 cells.Immunohistochemical staining was applied to test the expression of CTGF and the expression level of CTGF mRNA was assayed by RT-PCR.Results Basal levels of CTGF protein were observed in cultured HLF-02.The expression of CTGF,CTGF mRNA,p-Smad2 and p-ERK 1/2,in HLF-02 cells were stimulated significantly（P〈0.01） by 1 ng/mL TGF-β 1 treatment.The increase of CTGF protein was inhibited significantly by pre-incubation with Staurosporine（SP） and PD98059（P〈0.01）.And matrine could markedly inhibited the up-regulation of CTGF, CTGF mRNA,p-Smad2 and p-ERK 1/2（P〈0.05or P〈0.01） in a concentration-dependant manner.Conclusion CTGF,CTGF mRNA,p-Smad2 and p-ERK 1/2 of HLF-02 cells could be significantly up-regulated after treated with TGF-β 1,and these increases could be inhibited significantly by matrine.This implied that matrine inhibit the expression of CTGF and CTGF mRNA induced by TGF-β 1 through the Smad2 and ERK signal transduction pathways.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2009年第6期994-999,共6页
Journal of Sichuan University(Medical Sciences)
基金
四川省科技厅攻关项目(0040205301259)资助
