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c-jun氨基末端激酶(JNK)信号转导通路活化的调节在矽肺纤维化中的作用.方法选用非暴露式气管灌注法制作大鼠矽肺模型.60只大鼠随机分为对照4周组、对照8周组、矽肺模型4周组、矽肺模型8周组、AcSDKP治疗组、AcSDKP预防组,每组10只.采用对二甲氨基苯甲醛显色法检测肺组织中羟脯氨酸含量,免疫印迹法检测肺组织内转化生长因子(TGF)-β1、磷酸化-JNK、JNK、c-jun蛋白的表达.培养新生大鼠肺成纤维细胞,第4代细胞用于实验,分为对照组、TGF-β1刺激组、SP600125干预组、AcSDKP干预组.免疫细胞化学法检测磷酸化-JNK和c-jun蛋白的分布,免疫印迹法检测Ⅰ型、Ⅲ型胶原表达.结果 AcSDKP治疗组大鼠肺组织中TGF-β1、磷酸化-JNK、c-jun蛋白表达和羟脯氨酸含量分别是矽肺模型4周组的70.60％、78.03％、79.85％和71.28％,分别是矽肺模型8周组的77.99％、66.73％、69.94％和64.82％；AcSDKP预防组大鼠肺组织中TGF-β1、磷酸化-JNK、c-jun蛋白表达和羟脯氨酸含量分别为矽肺模型8周组的84.56％、61.18％、64.73％和74.96％,差异均有统计学意义(P＜0.05).AcSDKP干预组大鼠肺成纤维细胞磷酸化-JNK和c-jun蛋白的表达分别是TGF-β1刺激组的54.59％和55.56％,AcSDKP干预组大鼠肺成纤维细胞Ⅰ型和Ⅲ型胶原蛋白的表达分别是TGF-β1刺激组的79.9％和84.4％,差异均有统计学意义(P＜0.05).结论 AcSDKP可能通过阻制TGF-β1介导的JNK信号转导通路的活化,抑制间质胶原的沉积,进而发挥其抗矽肺纤维化的作用.
목적 탐토N-을선기-사안선-천문동선-뢰안선-포안산(AcSDKP)대c-jun안기말단격매(JNK)신호전도통로활화적조절재석폐섬유화중적작용.방법 선용비폭로식기관관주법제작대서석폐모형.60지대서수궤분위대조4주조、대조8주조、석폐모형4주조、석폐모형8주조、AcSDKP치료조、AcSDKP예방조,매조10지.채용대이갑안기분갑철현색법검측폐조직중간포안산함량,면역인적법검측폐조직내전화생장인자(TGF)-β1、린산화-JNK、JNK、c-jun단백적표체.배양신생대서폐성섬유세포,제4대세포용우실험,분위대조조、TGF-β1자격조、SP600125간예조、AcSDKP간예조.면역세포화학법검측린산화-JNK화c-jun단백적분포,면역인적법검측Ⅰ형、Ⅲ형효원표체.결과 AcSDKP치료조대서폐조직중TGF-β1、린산화-JNK、c-jun단백표체화간포안산함량분별시석폐모형4주조적70.60％、78.03％、79.85％화71.28％,분별시석폐모형8주조적77.99％、66.73％、69.94％화64.82％；AcSDKP예방조대서폐조직중TGF-β1、린산화-JNK、c-jun단백표체화간포안산함량분별위석폐모형8주조적84.56％、61.18％、64.73％화74.96％,차이균유통계학의의(P＜0.05).AcSDKP간예조대서폐성섬유세포린산화-JNK화c-jun단백적표체분별시TGF-β1자격조적54.59％화55.56％,AcSDKP간예조대서폐성섬유세포Ⅰ형화Ⅲ형효원단백적표체분별시TGF-β1자격조적79.9％화84.4％,차이균유통계학의의(P＜0.05).결론 AcSDKP가능통과조제TGF-β1개도적JNK신호전도통로적활화,억제간질효원적침적,진이발휘기항석폐섬유화적작용.
Objective To investigate the regulatory effect of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) on the activation of c-jun N-terminal kinase (JNK) signal transduction pathway and its role in silicotic fibrosis.Methods A rat model of silicosis was developed by intratracheal instillation.Sixty rats were randomly divided into 4-week control group (n=10),8-week control group (n=10),4-week silicosis model group (n=10),8-week silicosis model group (n=10),AcSDKP treatment group (n=10),and AcSDKP prevention group (n=10).The content of hydroxyproline in lung tissue was measured using a p-dimethylaminobenzaldehyde reagent; the expression levels of transforming growth factor (TGF)-beta 1 (TGF-β1),phospho-JNK,JNK,and c-jun in lung tissue were measured by Western blot.The lung fibroblasts from neonatal rats were cultured,and the 4th generation of cells were used in the experiment; these cells were divided into control group,TGF-β1 stimulation group,SP600125 intervention group,and AcSDKP intervention group.The distributions of phospho-JNK and c-jun in lung fibroblasts were observed by immunocytochemistry; the expression levels of type Ⅰ collagen and type Ⅲ collagen in lung fibroblasts were measured by Western blot.Results The expression levels of TGF-β13 phospho-JNK,and c-jun and the content of hydroxyproline in the AcSDKP treatment group were 70.60％,78.03％,79.85％,and 71.28％,respectively,of those in the 4-week silicosis model group (P＜0.05) and 77.99％,66.73％,69.94％,and 64.82％,respectively,of those in the 8-week silicosis model group (P＜0.05); the expression levels of TGF-β1,phospho-JNK,and c-jun and the content of hydroxyproline in the AcSDKP prevention group were 84.56％,61.18％,64.73％,and 74.96％,respectively,of those in the 8-week silicosis model group (P＜0.05).The expression levels of phospho-JNK and c-jun in the AcSDKP intervention group were 54.59％ and 55.56％,respectively,of those in the TGF-β1 stimulation group; the expression levels of type Ⅰ collagen and type Ⅲ collagen in the AcSDKP intervention group were 79.9％ and 84.4％,respectively,of those in the TGF-β1 stimulation group (P＜0.05).Conclusion AcSDKP exerts anti-silicotic fibrosis effect probably by inhibiting the activation of JNK signal transduction pathway mediated by TGF-β1 and the deposition of interstitial collagen.