四川大学学报（医学版）
四川大學學報（醫學版）
사천대학학보（의학판）
JOURNAL OF SICHUAN UNIVERSITY(MEDICAL SCIENCE EDITION)
2009年
6期
994-999
,共6页
人肺成纤维细胞%转化生长因子-β_1%结缔组织生长因子%苦参碱
人肺成纖維細胞%轉化生長因子-β_1%結締組織生長因子%苦參堿
인폐성섬유세포%전화생장인자-β_1%결체조직생장인자%고삼감
Human lung fibroblasts%Transforming growth factor-β_1%Connective tissue growth factor%Matrine
目的 研究苦参碱对肺成纤维细胞转化生长因子-β_1(TGF-β_1)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-β_1、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(ERK_(1/2))抑制剂PD98059、苦参碱(Matrine,Mat)作用对结缔组织生长因子(CTGF)、p-Smad2、p-ERK_(1/2)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CTGF mRNA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-β_1可使CTGF、CTGF mRNA、p-Smad2、p-ERK_(1/2)表达水平增强(P<0.01);使用SP及PD98059预处理细胞可以抑制TGF-β_1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-β_1刺激的CTGF、CTGF mRNA、p-Smad2、p-ERK_(1/2)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-β_1可使CTGF蛋白及CTGF mRNA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-β_1诱导的CTGF表达.
目的 研究苦參堿對肺成纖維細胞轉化生長因子-β_1(TGF-β_1)信號轉導途徑的影響.方法 以人肺成纖維細胞繫(HLF-02)為研究對象,用Western blot法檢測TGF-β_1、絲/囌氨痠激酶抑製劑星形孢菌素(Staurosporine,SP)和細胞外信號調節激酶(ERK_(1/2))抑製劑PD98059、苦參堿(Matrine,Mat)作用對結締組織生長因子(CTGF)、p-Smad2、p-ERK_(1/2)蛋白錶達的影響,免疫組織化學方法檢測CTGF蛋白,RT-PCR技術檢測CTGF mRNA的錶達.結果 體外培養的HLF-02錶達基礎水平的CTGF蛋白,1 ng/mL TGF-β_1可使CTGF、CTGF mRNA、p-Smad2、p-ERK_(1/2)錶達水平增彊(P<0.01);使用SP及PD98059預處理細胞可以抑製TGF-β_1刺激的CTGF的錶達增加(P<0.01).使用苦參堿預處理細胞可以抑製TGF-β_1刺激的CTGF、CTGF mRNA、p-Smad2、p-ERK_(1/2)蛋白的錶達(P<0.05或P<0.01),且與苦參堿濃度呈負相關(r分彆為0.845,0.900,0.789,0.942;P<0.01).結論 TGF-β_1可使CTGF蛋白及CTGF mRNA的錶達明顯增彊,可能是通過Smad2和ERK途徑促進CTGF錶達.苦參堿可能是通過Smad2和ERK通路在基因及蛋白水平抑製TGF-β_1誘導的CTGF錶達.
목적 연구고삼감대폐성섬유세포전화생장인자-β_1(TGF-β_1)신호전도도경적영향.방법 이인폐성섬유세포계(HLF-02)위연구대상,용Western blot법검측TGF-β_1、사/소안산격매억제제성형포균소(Staurosporine,SP)화세포외신호조절격매(ERK_(1/2))억제제PD98059、고삼감(Matrine,Mat)작용대결체조직생장인자(CTGF)、p-Smad2、p-ERK_(1/2)단백표체적영향,면역조직화학방법검측CTGF단백,RT-PCR기술검측CTGF mRNA적표체.결과 체외배양적HLF-02표체기출수평적CTGF단백,1 ng/mL TGF-β_1가사CTGF、CTGF mRNA、p-Smad2、p-ERK_(1/2)표체수평증강(P<0.01);사용SP급PD98059예처리세포가이억제TGF-β_1자격적CTGF적표체증가(P<0.01).사용고삼감예처리세포가이억제TGF-β_1자격적CTGF、CTGF mRNA、p-Smad2、p-ERK_(1/2)단백적표체(P<0.05혹P<0.01),차여고삼감농도정부상관(r분별위0.845,0.900,0.789,0.942;P<0.01).결론 TGF-β_1가사CTGF단백급CTGF mRNA적표체명현증강,가능시통과Smad2화ERK도경촉진CTGF표체.고삼감가능시통과Smad2화ERK통로재기인급단백수평억제TGF-β_1유도적CTGF표체.
Objective To investigate the effect of matrine on the signal transduction pathway of transforming growth factor-pi (TGF-β_1). Methods The effect of TGF-β_1, 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-β1, 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.
