CAJ | 학술논문

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(ERK1/2)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1.0 ml/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1 ml/只(SiO250mg/ml);抗纤维化治疗组(治疗组):支气管内灌注SiO2混悬液1 ml/只,4周后给予AcSDKP 800μg·kg-1·d-1,并持续至第8周处死;预防治疗组:给予AcSDKP 800μg·kg-1·d-148 h后,支气管内灌注SiO2混悬液1 ml/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%、78.91%,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.
목적 탐토N-을선기-사안선-천문동선-뢰안선-포안산(AcSDKP)대석폐대서섬유화폐조직중c-Raf、세포외신호조절격매1/2(ERK1/2)화전화생장인자(TGF)-β1표체적영향,이탐토AcSDKP대Ras-Raf-ERK1/2신호전도통로조절적작용.방법 선용비폭로식기관관주법복제대서석폐모형,60지대서수궤분위6조,매조10지.대조1조(4주후처사)화대조2조(8주후처사):지기관내균관주생리염수1.0 ml/지;석폐모형1조(4주후처사)화석폐모형2조(8주후처사):지기관내관주SiO2혼현액1 ml/지(SiO250mg/ml);항섬유화치료조(치료조):지기관내관주SiO2혼현액1 ml/지,4주후급여AcSDKP 800μg·kg-1·d-1,병지속지제8주처사;예방치료조:급여AcSDKP 800μg·kg-1·d-148 h후,지기관내관주SiO2혼현액1 ml/지,8주후처사.채용면역조직화학법화면역인적(Western blot)법대석폐대서폐내c-Raf、린산화-c-Raf、ERK1/2、린산화-ERK1/2화TGF-β1적단백표체진행검측.결과 여상응적대조조상비,석폐모형조대서폐조직내린산화-c-Raf、린산화-ERK1/2화TGF-β1단백표체균증가;여상응석폐모형조상비,치료조대서폐조직내린산화-c-Raf、린산화-ERK1/2화TGF-β1단백표체균명현강저,기중,치료조린산화-c-Raf、린산화-ERK1/2화TGF-β1단백표체분별위석폐모형1조적52.25%、51.72%、67.74%,위석폐모형2조적49.37%、55.76%、65.63%;예방치료조단백표체분별위석폐모형2조적54.64%、55.76%、78.91%,차이균유통계학의의(P＜0.05).이각조간비교c-Raf、ERK1/2단백표체무명현개변.결론 AcSDKP가능시통과억제TGF-β1개도적Ras/Raf/ERK1/2신호통로발휘길항석폐섬유화적작용.
Objective To investigate the effect of N-acetyl-seryl-aspartyl-lysyl-proline(AcSDKP) on the expressions of c-Raf, ERK 1/2 and TGF-β1 in the lung of rats with silicosis, thus to investigate the regulating of AcSDKP on the Ras-Raf-ERK 1/2 signal transduction pathway. Methods Rats were instilled with silica through trachea as silicotic models and administered AcSDKP in the experiment. Rats were divided into 6groups randomly, 10 rats in each group: Control 1 and 2 of silicotic 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 intratracheally instilled with 1ml silica suspension and was killed after 4 or 8 weeks; Anti-fibrosis treatment of AcSDKP: after each rat was intratracheally instilled with lml silica suspension for 4 weeks, AcSDKP 800 μg ·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 μg ·kg-1 ·d-1 was administered into every rat for 48 hours, each rat was intratracheally 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 phosphoc-Raf, phospho-ERK1/2 and TGF-β1 increased in the lung tissue of the silicotic 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 silicotic model 2; In preventing fibrosis treatment of AcSDKP group, expressions of phospho-c-Raf, phospho-ERK 1/2 and TGF-β1 decreased to 54.64%, 55.76% and 78.91%compared with those of the silicotic 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-silicotic fibrosis by blocking the TGF-β-induced Ras-Raf-ERK 1/2 signal transduction pathway.