CAJ | 학술논문

目的观察实验性大鼠肝纤维化形成过程中,肝组织瘦素、转化生长因子(TGF)-β1水平的动态变化,探讨瘦素在肝纤维化发生过程中可能的作用及机制.方法以38只SD大鼠为实验对象,于实验开始时处死6只作为正常对照组,剩余32只皮下注射60%四氯化碳复制肝纤维化模型,分别于3、6、9、12周各处死8只.RT-PCR检测大鼠肝组织中瘦素、瘦素功能性受体(Ob-Rb)、TGF-β1及其受体(TGF-βR Ⅱ)的mRNA表达水平.Western印迹法榆测肝组织中Smad3、Smad7蛋白水平.免疫组化法检测肝组织中瘦素、Ob-Rb及TGF-β1的表达和定位.结果正常肝组织中有少量瘦素、Ob-Rb、TGF-β1,TGF-βRⅡ的mRNA表达(分别为0.43±0.45、0,57±0.21、0.19±0.12、0.30±0.22),随着肝纤维化的逐渐形成,其表达量逐渐递增,至12周时分别为1.27±0.19、1.70±0.29、1.70±1.00、2.14±1.02,差异有统计学意义(P＜0.05).随着肝纤维化的形成,Smad3表达量逐渐递增,由0周时的0.44±0.24增至12周时的1.75±1.05,差异有统计学意义(P＜0.05);Smad7表达量逐渐减少,由0周时的1.35±0.12降至12周时的0.74±1.21,差异有统计学意义(P＜0.05).随着肝纤维化的形成,瘦素、Ob-Rb及TGF-β1表达水平增高(P＜0.01).结论随着肝纤维化的形成,肝脏中瘦素、Ob-Rb基因及蛋白表达增加,有促肝纤维化作用.其机制可能与瘦素通过直接或间接上调TGF-61、TGF-βR Ⅱ及Smad3表达、下调Smad7表达有关.
목적 관찰실험성대서간섬유화형성과정중,간조직수소、전화생장인자(TGF)-β1수평적동태변화,탐토수소재간섬유화발생과정중가능적작용급궤제.방법 이38지SD대서위실험대상,우실험개시시처사6지작위정상대조조,잉여32지피하주사60%사록화탄복제간섬유화모형,분별우3、6、9、12주각처사8지.RT-PCR검측대서간조직중수소、수소공능성수체(Ob-Rb)、TGF-β1급기수체(TGF-βR Ⅱ)적mRNA표체수평.Western인적법유측간조직중Smad3、Smad7단백수평.면역조화법검측간조직중수소、Ob-Rb급TGF-β1적표체화정위.결과 정상간조직중유소량수소、Ob-Rb、TGF-β1,TGF-βRⅡ적mRNA표체(분별위0.43±0.45、0,57±0.21、0.19±0.12、0.30±0.22),수착간섬유화적축점형성,기표체량축점체증,지12주시분별위1.27±0.19、1.70±0.29、1.70±1.00、2.14±1.02,차이유통계학의의(P＜0.05).수착간섬유화적형성,Smad3표체량축점체증,유0주시적0.44±0.24증지12주시적1.75±1.05,차이유통계학의의(P＜0.05);Smad7표체량축점감소,유0주시적1.35±0.12강지12주시적0.74±1.21,차이유통계학의의(P＜0.05).수착간섬유화적형성,수소、Ob-Rb급TGF-β1표체수평증고(P＜0.01).결론 수착간섬유화적형성,간장중수소、Ob-Rb기인급단백표체증가,유촉간섬유화작용.기궤제가능여수소통과직접혹간접상조TGF-61、TGF-βR Ⅱ급Smad3표체、하조Smad7표체유관.
Objective To observe the dynamic changes of leptin and transforming growth factor (TGF)-β1 in formation of liver fibrosis in rats,and the role of leptin in liver fibrosis.Methods Six rats were served as normal control(0 week)and killed at the beginning of the study.Thirty-two rats were subcutaneously injected with 60%CCl4 to establish fibrotie models.The rats were than sacrificed at 3,6,9 and 12 weeks with eight each.Expressions of leptin mRNA,Ob-Rb mRNA,TGF-β1 mRNA and TGF-βRⅡmRNA in liver were detected by reverse-transcription polymerase chain reaction(RT-PCR).Expression of Smad3 and Smad7 proteins were determined by Western blot.The expression and loealization of leptin,Ob-Rb and TGF-β1 in liver tissue were detected by immunohistochemistry.Results The expression of leptin mRNA,Ob-Rb mRNA,TGF-β1 mRNA and TGF-βR Ⅱ mRNA in 0 week were 0.43±0.45,0.57±0.21,0.19±0.12 and 0.30±0.22,respectively,and increased to 1.27±0.19,1.70±0.29,1.70±1.00 and 2.14±1.02 at 12 weeks(P＜0.05).At the same times the level of Smad3 increased from 0.44±0.24 at 0 week to 1.75±1.05 at 12 weeks.While the expression of Smad7 was decreased from 1.35±0.12 at 0 week to 0.74±1.21 at 12 weeks(P＜0.05).The expressions of leptin,Ob-Rb and TGF-β1 were increased with the formation of the fibrosis(P＜0.01).Conclusions With the development of liver fibrosis,the expressions of leptin,Ob-Rb mRNA and protein are increased and improved the formation of the fibrosis.Its mechanisms may be correlated with up-regulation of TGF-β1,TGF-βR Ⅱ and Smda3 expressions and down-regulaton of Smad7 expression by leptin.