摘要
目的探讨N-乙酰基-丝氨酰-天门冬酰-赖氨酰-脯氨酸（AcSDKP）对矽肺大鼠纤维化肺组织中c—Raf、细胞外信号调节激酶1／2（ERKl／2）和转化生长因子（TGF）-β1表达的影响，以探讨AcSDKP对Ras—Raf-ERK1／2信号转导通路调节的作用。方法选用非暴露式气管灌注法复制大鼠矽肺模型，60只大鼠随机分为6组，每组10只。对照1组（4周后处死）和对照2组（8周后处死）：支气管内均灌注生理盐水1．0ml/只；矽肺模型1组（4周后处死）和矽肺模型2组（8周后处死）：支气管内灌注SiO2混悬液1ml／只（SiO2 50mg／ml）；抗纤维化治疗组（治疗组）：支气管内灌注SiO2混悬液1ml／只，4周后给予AcSDKP800ug·kg-1·d-1，并持续至第8周处死；预防治疗组：给予AcSDKP800ug·kg-1·d-1 48h后，支气管内灌注SiO2混悬液1ml／只，8周后处死。采用免疫组织化学法和免疫印迹（Western blot）法对矽肺大鼠肺内c—Raf、磷酸化-c-Raf、ERK1／2、磷酸化-ERK1／2和TGF—β1的蛋白表达进行检测：结果与相应的对照组相比，矽肺模型组大鼠肺组织内磷酸化-c-Raf、磷酸化-ERK1／2和TGF-β1蛋白表达均增加；与相应矽肺模型组相比，治疗组大鼠肺组织内磷酸化-c-Raf、磷酸化-ERK1／2和TGF—β1蛋白表达均明显降低，其中，治疗组磷酸化-c-Raf、磷酸化-ERK1／2和TGF—β1蛋白表达分别为矽肺模型1组的52．25％、51．72％、67．74％，为矽肺模型2组的49．37％、55．76％、65．63％；预防治疗组蛋白表达分别为矽肺模型2组的54．64％、55．76％、78．9／％，差异均有统计学意义（P〈0．05）。而各组间比较c—Raf、ERK1／2蛋白表达无明显改变。结论AcSDKP可能是通过抑制TGF—β1介导的Ras／Raf／ERK1／2信号通路发挥拮抗矽肺纤维化的作用。
Objective To investigate the effect of N-acetyl-seryl-aspartyl-lysyl-proline （AcSDKP） on the expressions of c-Raf, ERK1/2 and TGF-β1 in the lung of rats with silicosis, thus to investigate the regulat- ing of AeSDKP on the Ras-Raf-ERK1/2 signal transduction pathway. Methods Rats were instilled with silica through trachea as silicotic models and administered AeSDKP in the experiment. Rats were divided into 6 groups randomly, 10 rats in each group： Control 1 and 2 of silicotie model： each rat was intratracheally instilled with 1.0 ml normal sodium and was killed after 4 or 8 weeks; Silicotic model 1 and Silicotic model 2： each rat was intratraeheally instilled with lml silica suspension and was killed after 4 or 8 weeks; Anti-fibrosis treatment of AeSDKP： after each rat was intratracheally instilled with lml siliea suspension for 4 weeks, AeSDKP 800 ug·kg-1·d-1 was administered into every, rat and rats were killed at the eighth week; Preventing fibrosis treatment of AcSDKP： after AcSDKP 800 ug·kg-1·d -1 was administered into every rat for 48 hours, each rat was intratraeheally instilled with 1.0 ml silica suspension and rats were killed at the eighth week. The expression of c-Raf, phospho-c-Raf, ERK1/2, phospho-ERK1/2 and TGF-β1 was measured by immunohistochemistry and western blot assay. Results Compared with the corresponding control groups,the expressions of phospho- c-Raf, phospho-ERK1/2 and TGF-β1 increased in the lung tissue of the silieotic models. Compared with the corresponding model groups, after administration AcSDKP, the expressions of phospho-c-Raf, phospho-ERK1/2 and TGF-β1 in the lung tissue reduced obviously. In anti-fibrosis treatment of AcSDKP group, expressions of phospho-c-Raf. phospho-ERK1/2 and TGF-β1 decreased to 52.25%. 51.72% and 67.74% compared with those of the silicotic model 1. and expressions of phospho-c-Raf. phospho-ERK1/2 and TGF-β1 decreased to 49.37%, 55.76%, 65.63% compared with those of the silieotic model 2： In preventing fibrosis treatment of AcSDKP group, expressions of phuspho-c-Raf. phospho-ERK1/2 and TGF-β1 decreased to 54.64%. 55.76% and 78.91% compared with those of the silicotie model 2 （P〈0.05） while the expressions of c-Raf and ERK1/2 were not different significantly among each groups. Conclusion AcSDKP possibly plays an important role in anti-silicotie fibrosis by blocking the TGF-β-induced Ras-Raf-ERK 1/2 signal transduction pathway.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
2010年第10期760-765,共6页
Chinese Journal of Industrial Hygiene and Occupational Diseases
基金
基金项目：河北省科技支撑计划项目（402249546）
中华人民共和国人事部留学人员科技活动资助项目（国人厅发[2006]164号）
河北省自然科学基金资助项目（C2005000807）
唐山市新药基础研究重点实验室资助项目（04362001B-9）
