摘要
目的 :研究缺氧时大鼠肺动脉平滑肌细胞 (PASMC)诱导型血红素氧合酶 (HO 1 ) /一氧化碳 (CO)系统的变化及其对胶原合成的影响 ,探讨血红素氧合酶 /一氧化碳 (HO/CO)系统对缺氧性肺血管重建中胶原代谢的作用及其机制。方法 :原代培养的大鼠PASMC ,用分光光度计检测PASMC培养液中CO相对含量的变化 ,Westernblot检测PASMCHO 1和转化生长因子 β3 (TGF β3 )表达的变化 ,用免疫细胞化学法分别观察PASMCHO 1、TGF β3 、Ⅰ型胶原蛋白表达的变化 ,原位杂交法检测Ⅰ型前胶原mRNA表达的变化。结果 :缺氧 2 4h诱导PASMC表达TGF β3 、Ⅰ型胶原蛋白和mRNA ,与对照组比较缺氧使HO 1蛋白表达增加 6 7.4 5 % (P <0 .0 1 )、CO含量增加35 .4 1 % (P <0 .0 5 )。ZnPP(HO 1抑制剂 )使缺氧 2 4h大鼠PASMC的CO含量降低 7.88% (P <0 .0 1 )、HO 1蛋白表达减少 2 3.9% (P <0 .0 5 )、TGF β3 蛋白表达增高 393% (P <0 .0 1 )、Ⅰ型胶原蛋白和mRNA表达均增加。Hemin(HO 1诱导剂 )使缺氧 2 4h大鼠PASMC的CO含量增加 8.83% (P <0 .0 1 )、HO 1表达增高 1 0 5 % (P <0 .0 5 )、TGF β3 蛋白表达降低 6 8.1 2 % (P <0 .0 1 ) ,Ⅰ型胶原蛋白和mRNA表达均减少。结论 :缺氧刺激下大鼠PASMCHO/CO系统上调 ,内源性CO能够抑制Ⅰ型胶原?
Objective: To explore changes of expression of heme oxygenase (HO) and production of endogenous carbon monoxide in hypoxic pulmonary artery smooth muscle cells (PASMCs) of rats and study their effects on type Ⅰcollagen under hypoxia. Methods: PASMCs of rats were cultured in vitro and incubated under hypoxic condition for 24 hours. The relative content of CO released into medium by PASMCs was detected using spectrophotometry. Western blot was applied to assay the expressions of HO 1 and transforming growth factor β 3 (TGF β 3). The expressions of HO 1, TGF β 3 and typeⅠcollagen were detected by immunohistochemical assay. The expressions of procollagen type Ⅰwere detected by in situ hybridization. Results: Hypoxia induced the expression of TGF β 3, typeⅠcollagen and mRNA in rat PASMCs, elevated the expression of HO 1 and CO content compared to controls by 67.45% ( P <0.01) and 35.41%( P <0.05), respectively. In the hypoxic PASMCs treated with protoporphyrin IX (ZnPP)(10 μmol·L -1 ), an inhibitor of HO, the expression of HO 1 and the CO content were 23.9%( P < 0.05), 7.88% ( P <0.01) lower than those of hypoxic PASMCs, respectively. ZnPP promoted the expression of TGF β 3 (393%, P <0.01), typeⅠcollagen protein and mRNA in hypoxic PASMCs. Hemin (20 μmol·L -1 ), an inducer of HO, caused an increase in the expression of HO 1 and CO content in hypoxic rat PASMCs by 105% ( P <0.05), 8.83% ( P <0.01) compared to hypoxic PASMCs, respectively. Hemin inhibited the expression of TGF β 3 by 68.12%( P <0.01), and typeⅠcollagen protein and mRNA in hypoxic PASMCs. Conclusion: HO/CO system is up regulated by hypoxia and endogenous CO may play an important role in decreasing collagen synthesis and inhibiting the expression of TGF β 3 in hypoxic rat PASMCs.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2003年第6期571-575,共5页
Journal of Peking University:Health Sciences
基金
国家自然科学基金 (30 0 70 796)
国家重点基础研究项目基金 (G2 0 0 0 0 5690 5)
北京市自然科学基金 (70 1 2 0 2 1 )资助~~
关键词
缺氧
肺动脉
平滑肌细胞血红素氧合酶
一氧化碳
Ⅰ型胶原
病理学
Carbon monoxide
Pulmonary artery
Cell hypoxia
Collagen type Ⅰ
Muscle, smooth, vascular/physiopathol
