博来霉素诱导培养的人胎肺成纤维细胞中赖氨酰氧化酶的上调。
Bleomycin induces upregulation of lysyl oxidase in cultured human fetal lung fibroblasts.
机构信息
Department of Pharmacology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510089, China.
出版信息
Acta Pharmacol Sin. 2010 May;31(5):554-9. doi: 10.1038/aps.2010.44. Epub 2010 Apr 26.
AIM
To investigate the mechanism of bleomycin (BLM)-induced pulmonary fibrosis.
METHODS
Cultured human fetal lung fibroblast (HLF) cells were exposed to bleomycin (BLM) at 0-30 microg/mL for 24 h. Western blot analysis was used to detect lysyl oxidase (LO) protein expression. Real-time RT-PCR was used to detect LO mRNA level. LO catalytic activity was measured using diaminopentane as a substrate and Amplex red as a hydrogen peroxide probe. Copper (Cu) concentration was detected by flame atomic absorption spectrophotometry.
RESULTS
Exposure of HLF cells to BLM at 10 microg/mL and 30 microg/mL increased LO catalytic activity to 130% and 158% of the control in the conditioned media. The expression of LO mRNA was increased to 5.5-fold of the control in HLF cells exposure to BLM at 3 microg/mL. BLM at 3 microg/mL also increased the expression of 46 kDa preproLO, 50 kDa proLO and 32 kDa mature LO to 219%, 130%, and 135% of the control, respectively. The Cu concentrations in conditioned media of cultured HLF cells exposed to BLM (10 and 30 microg/mL) were increased significantly to 1.48 and 2.46-fold of the control, respectively.
CONCLUSION
Bleomycin induces upregulation of LO in cultured human fetal lung fibroblasts, which may be the mechanism of bleomycin-induced pulmonary fibrosis.
目的
探讨博来霉素（BLM）诱导肺纤维化的机制。
方法
将培养的人胎肺成纤维细胞（HLF）暴露于 0-30μg/ml 的博来霉素（BLM）中 24 小时。采用 Western blot 分析检测赖氨酰氧化酶（LO）蛋白表达。实时 RT-PCR 检测 LO mRNA 水平。以二氨基戊烷为底物，Amplex red 为过氧化氢探针，测定 LO 催化活性。火焰原子吸收分光光度法检测铜（Cu）浓度。
结果
HLF 细胞暴露于 10μg/ml 和 30μg/ml 的 BLM 时，LO 催化活性分别增加至对照的 130%和 158%。HLF 细胞暴露于 3μg/ml 的 BLM 时，LO mRNA 的表达增加至对照的 5.5 倍。BLM 还可将 46 kDa 前 proLO、50 kDa proLO 和 32 kDa 成熟 LO 的表达分别增加至对照的 219%、130%和 135%。暴露于 BLM（10 和 30μg/ml）的培养 HLF 细胞的条件培养基中的 Cu 浓度分别显著增加至对照的 1.48 倍和 2.46 倍。
结论
博来霉素诱导培养的人胎肺成纤维细胞中 LO 的上调，这可能是博来霉素诱导肺纤维化的机制。
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