目的探讨降低表皮生长因子受体（EGFR）在肺内的表达对博莱霉素致小鼠肺纤维化的影响。方法将40只4-6周龄C57BLB／c雄性小鼠按简单随机法分为正常对照组（气管滴入PBS）、纤维化组（气管滴入博莱霉素3mg／kg）、RNAi组（气管滴入博莱霉...目的探讨降低表皮生长因子受体（EGFR）在肺内的表达对博莱霉素致小鼠肺纤维化的影响。方法将40只4-6周龄C57BLB／c雄性小鼠按简单随机法分为正常对照组（气管滴入PBS）、纤维化组（气管滴入博莱霉素3mg／kg）、RNAi组（气管滴入博莱霉素3mg／kg＋气管滴入siR-NA20μl）和RNAi阴性对照组（气管滴入博莱霉素3mg／kg＋气管滴人siRNA阴性对照20μl）。实验第10天处死小鼠，收获肺组织，检测羟脯氨酸含量。肺组织切片行HE染色观察肺组织病理改变，采用逆转录一聚合酶链反应（RT-PCR）法检测EGFR的表达。Westernblot检测EGFR、磷酸化EGFR表达。结果RNAi组与纤维化组比较，肺组织EGFRmRNA表达（0．31±0．05 vs 0．75±0．08，P〈0．01）和EGFR蛋白表达（1．53±O．47vs2．56±0．37，P〈0．01）均显著下降；肺病理损伤较纤维化组减轻，肺羟脯氨酸含量显著减少（543．00±25．89vs900．73±31．77，P〈0．01）；磷酸化EGFR蛋白表达亦较纤维化组明显减少（1．78±0．35 vs 2．84±0．51，P〈0．01）。结论EGFRRNAi抑制了EGFR活化，减轻了博莱霉素诱导的肺纤维化改变。展开更多
Objective To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 （ TGF-β1...Objective To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 （ TGF-β1 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and matrix metalloproteinase-13 （ MMP-13 ） in lung tissue. Methods Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone （25-800 mg · kg^-l · d^-1 ）, dexamethasone （3 mg/kg）, or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg · kg^-1 · d^-1 at 7 days or 14 daYs after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in luffg tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxypro- line. Expression of proteins of TGF-β1 TIMP-1, and MMP-13 were detected by Western blotting. Results At doses of 25, 50, and 100 mg· kg^- 1 · d ^- 1, pirfenidone had significant anti-fibrotic effects for bleomy- cin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg · kg^-1 ·d^ -1（ HE： P 〈 0. 01, P 〈 0.01, and P = 0.064; sirius red： P 〈0.05, P 〈 0.01, and P 〈 0.05 ; hydroxyproline： P = 0.595, P 〈 0.01, and P = 0.976）. Pirfenidone at a dosage of 50 mg · kg^- l · d^-1 inhibited protein expression of TGF-131 and TIMP-1 in lung tissue in the early phase （0.79 and 0.75 times of control group）, but had no effect on ex- nr^eelnn nf MMP-13. Conclusion Low dose pirfenidone, especially at dosage of 50 mg · kg^-1 · d^-1, has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-131 and TIMP-β1 in lung tissue.展开更多
文摘目的探讨降低表皮生长因子受体（EGFR）在肺内的表达对博莱霉素致小鼠肺纤维化的影响。方法将40只4-6周龄C57BLB／c雄性小鼠按简单随机法分为正常对照组（气管滴入PBS）、纤维化组（气管滴入博莱霉素3mg／kg）、RNAi组（气管滴入博莱霉素3mg／kg＋气管滴入siR-NA20μl）和RNAi阴性对照组（气管滴入博莱霉素3mg／kg＋气管滴人siRNA阴性对照20μl）。实验第10天处死小鼠，收获肺组织，检测羟脯氨酸含量。肺组织切片行HE染色观察肺组织病理改变，采用逆转录一聚合酶链反应（RT-PCR）法检测EGFR的表达。Westernblot检测EGFR、磷酸化EGFR表达。结果RNAi组与纤维化组比较，肺组织EGFRmRNA表达（0．31±0．05 vs 0．75±0．08，P〈0．01）和EGFR蛋白表达（1．53±O．47vs2．56±0．37，P〈0．01）均显著下降；肺病理损伤较纤维化组减轻，肺羟脯氨酸含量显著减少（543．00±25．89vs900．73±31．77，P〈0．01）；磷酸化EGFR蛋白表达亦较纤维化组明显减少（1．78±0．35 vs 2．84±0．51，P〈0．01）。结论EGFRRNAi抑制了EGFR活化，减轻了博莱霉素诱导的肺纤维化改变。
基金Supported by National Ministry of Education Doctor Foundation of China(20020023045)
文摘Objective To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 （ TGF-β1 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and matrix metalloproteinase-13 （ MMP-13 ） in lung tissue. Methods Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone （25-800 mg · kg^-l · d^-1 ）, dexamethasone （3 mg/kg）, or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg · kg^-1 · d^-1 at 7 days or 14 daYs after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in luffg tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxypro- line. Expression of proteins of TGF-β1 TIMP-1, and MMP-13 were detected by Western blotting. Results At doses of 25, 50, and 100 mg· kg^- 1 · d ^- 1, pirfenidone had significant anti-fibrotic effects for bleomy- cin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg · kg^-1 ·d^ -1（ HE： P 〈 0. 01, P 〈 0.01, and P = 0.064; sirius red： P 〈0.05, P 〈 0.01, and P 〈 0.05 ; hydroxyproline： P = 0.595, P 〈 0.01, and P = 0.976）. Pirfenidone at a dosage of 50 mg · kg^- l · d^-1 inhibited protein expression of TGF-131 and TIMP-1 in lung tissue in the early phase （0.79 and 0.75 times of control group）, but had no effect on ex- nr^eelnn nf MMP-13. Conclusion Low dose pirfenidone, especially at dosage of 50 mg · kg^-1 · d^-1, has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-131 and TIMP-β1 in lung tissue.
