CAJ | 학술논문

目的观察kielin/chordin样蛋白(kcp)在肝纤维化小鼠模型中的动态表达及骨形成蛋白-7(BMP-7)干预对其表达的影响,探讨治疗肝纤维化的新靶点.方法健康雄性ICR小鼠50只,分为正常对照组(10只)、模型组(30只)和BMP-7干预组(10只),模型组再按造模不同时间点分4、8、12周3个亚组,每组10只.模型组小鼠皮下注射四氯化碳制备肝纤维化小鼠模型,BMP-7干预组在造模后第8周开始腹腔注射人重组BMP-7,持续4周.造模后各组小鼠下腔静脉采血测定血清ALT、AST和Alb;肝组织经HE和Masson染色后光学显微镜下观察各组病理变化;采用RT-PCR、免疫组织化学和Western印迹法分别检测各组kcp mRNA及蛋白表达情况.各组间均数比较采用单因素方差分析.结果模型组血清ALT、AST升高,A1b降低,12周时达峰值,BMP-7干预组有所缓解,各组间比较,差异有统计学意义(F值=23.501、34.600、16.244,均P＜0.05).正常对照组kcp mRNA和蛋白表达很低,模型组则逐渐升高,BMP-7干预组有所缓解,差异有统计学意义(F值=30.362、10.727,P＜0.01或0.05).kcp mRNA表达水平与转化生长因子β1( TGF-β1)、BMP-7 mRNA表达水平均呈正相关(r=0.760、0.769,均P＜0.05).结论 BMP-7能改善小鼠肝纤维化;kcp可能参与肝纤维化发生发展过程,有望成为肝纤维化治疗的新靶点.
목적 관찰kielin/chordin양단백(kcp)재간섬유화소서모형중적동태표체급골형성단백-7(BMP-7)간예대기표체적영향,탐토치료간섬유화적신파점.방법 건강웅성ICR소서50지,분위정상대조조(10지)、모형조(30지)화BMP-7간예조(10지),모형조재안조모불동시간점분4、8、12주3개아조,매조10지.모형조소서피하주사사록화탄제비간섬유화소서모형,BMP-7간예조재조모후제8주개시복강주사인중조BMP-7,지속4주.조모후각조소서하강정맥채혈측정혈청ALT、AST화Alb;간조직경HE화Masson염색후광학현미경하관찰각조병리변화;채용RT-PCR、면역조직화학화Western인적법분별검측각조kcp mRNA급단백표체정황.각조간균수비교채용단인소방차분석.결과 모형조혈청ALT、AST승고,A1b강저,12주시체봉치,BMP-7간예조유소완해,각조간비교,차이유통계학의의(F치=23.501、34.600、16.244,균P＜0.05).정상대조조kcp mRNA화단백표체흔저,모형조칙축점승고,BMP-7간예조유소완해,차이유통계학의의(F치=30.362、10.727,P＜0.01혹0.05).kcp mRNA표체수평여전화생장인자β1( TGF-β1)、BMP-7 mRNA표체수평균정정상관(r=0.760、0.769,균P＜0.05).결론 BMP-7능개선소서간섬유화;kcp가능삼여간섬유화발생발전과정,유망성위간섬유화치료적신파점.
Objective To observe the dynamic expressions of kielin/chordin-like protein (kcp) in mouse model of hepatic fibrosis and the effects of bone morphogenetic protein-7 (BMP-7)intervention on the expressions,and to explore a new target for treatment of fibrosis.Methods A total of 50 healthy male ICR mice were divided into three groups:control group(n=10) ; model group (n=30) and BMP-7 treatment group (n=10).The model group was further divided into three subgroups according to different time points:subgroups of 4,8 and 12 weeks with 10 mice in each subgroup.The mouse model of hepatic fibrosis was established by hypodermic injection of carbon tetrachloride(CCl4 ).The mice in BMP-7 treatment group began to receive human recombmant BMP- 7via intraperitoneal injection after 8 weeks of the first administration of CC14 and lasted for 4 weeks.The serum levels of alanine aminotransferase (ALT),aspartate transaminase (AST) and albumin (Alb) were detected.The pathological changes of liver were observed under optical microscope after HE and Masson staining.The dynamic expressions of kcp mRNA and protein of each group were detected by reverse transcription-polymerase chain reaction,immunohistochemistry and Western blot.The comparison of means among groups was done by univariate ANOVA.Results In model group,ALT and AST levels increased,while Alb level gradually decreased,and peaked at week 12.BMP-7treatment could reduce the changes,and there were significant differences among groups (F=23.501,34.600 and 16.244,respectively; all P＜0.05).In normal control group,the expressions of kcp mRNA and protein were low,while those in model group were gradually increased.BMP-7 treatment could achieve remission and the changes were all significantly different among groups (F=30.362 and 10.727,respectively; P＜0.01 or 0.05).The expression of kcp mRNA was positively correlated with levels of transforming growth factor (TGF)-β1 mRNA and BMP-7 mRNA (r=0.760 and 0.769,respectively; both P＜0.05).Conclusions BMP 7 can improve the hepatic fibrosis in mice.kcp may play an important role in the occurrence and development of liver fibrosis which is a potential therapeutic target for hepatic fibrosis.