CAJ | 학술논문

目的观察锌指蛋白(ZEB)1在大鼠肝纤维化模型中的表达情况及探讨重组ZEB1蛋白干预对纤维化程度的影响.方法 36只健康SD大鼠随机分为三组:对照组、模型组、ZEB1干预组.模型组与干预组均用二甲基亚硝胺(DMN)造模,每周前三天腹腔注射DMN,持续4周;之后干预组按4μg/kg剂量尾静脉注射重组ZEB1蛋白,隔天1次,而模型组继续腹腔注射DMN,各持续3周.对照组以等量等渗盐水代替DMN.实验结束后处死大鼠,留取肝组织经HE和Masson染色观察各组肝脏的病理学形态;用RT-PCR和Western blot检测肝组织转化生长因子(TGF)β 1和ZEB1的表达情况.计量资料采用单因素方差分析,两两比较采用LSD检验;计数资料采用Fisher确切概率法.结果病理学染色显示:模型组肝组织变性坏死,有明显纤维间隔形成,可见假小叶;ZEB1干预组与模型组相比,炎性坏死更加明显,纤维化程度加深,蓝色纤维条索更粗,两组肝纤维化程度的差异有统计学意义(x2=21.626,P＜0.01).RT-PCR和Western blot结果显示:ZEB1和TGF β 1在模型组中表达水平较正常组高,差异有统计学意义(P＜0.01);模型组与干预组比较,ZEB1和TGF β 1在干预组中表达水平更高,差异同样有统计学意义(P＜0.01). 结论 ZEB1可能通过参与TGF β/Smad通路促进大鼠肝纤维化的发展.
목적 관찰자지단백(ZEB)1재대서간섬유화모형중적표체정황급탐토중조ZEB1단백간예대섬유화정도적영향.방법 36지건강SD대서수궤분위삼조:대조조、모형조、ZEB1간예조.모형조여간예조균용이갑기아초알(DMN)조모,매주전삼천복강주사DMN,지속4주;지후간예조안4μg/kg제량미정맥주사중조ZEB1단백,격천1차,이모형조계속복강주사DMN,각지속3주.대조조이등량등삼염수대체DMN.실험결속후처사대서,류취간조직경HE화Masson염색관찰각조간장적병이학형태;용RT-PCR화Western blot검측간조직전화생장인자(TGF)β 1화ZEB1적표체정황.계량자료채용단인소방차분석,량량비교채용LSD검험;계수자료채용Fisher학절개솔법.결과 병이학염색현시:모형조간조직변성배사,유명현섬유간격형성,가견가소협;ZEB1간예조여모형조상비,염성배사경가명현,섬유화정도가심,람색섬유조색경조,량조간섬유화정도적차이유통계학의의(x2=21.626,P＜0.01).RT-PCR화Western blot결과현시:ZEB1화TGF β 1재모형조중표체수평교정상조고,차이유통계학의의(P＜0.01);모형조여간예조비교,ZEB1화TGF β 1재간예조중표체수평경고,차이동양유통계학의의(P＜0.01). 결론 ZEB1가능통과삼여TGF β/Smad통로촉진대서간섬유화적발전.
Objective To determine the role of zinc finger protein 1 (ZEB 1) in liver fibrosis and in regards to expression of the tumor growth factor-beta (TGFβ) signaling factor using a rat model system.Methods Sprague-Dawley rats were randomly divided into a normal (control) group,liver fibrosis (model) group and a liver fibrosis + therapy (ZEB1 intervention) group.The model group and the ZEB1 intervention group were given intraperitoneal injections of dimethylnitrosamine (DMN) for the first 3 days of each week over a 7-week period; starting at week 5,the ZEB 1 intervention group was started on a routine of every other day tail vein injections of recombinant ZEB1.During this 7-week period,the control group was given intraperitoneal injections of 0.9％ NaC1 alone on the DMN schedule.Liver tissues were collected for pathological examination (with hematoxylin-eosin and Masson staining) and for detection of TGFβ1 and ZEB 1 expression (by RT-PCR and western blotting).Measurement data were compared between groups using the single-factor analysis of variance test,followed by the least significant difference LSD test.Count data were analyzed by Fisher's exact test.Results The model group's liver tissues showed degeneration and necrosis,as well as obvious fibrous septa accompanied by pseudo lobules.The ZEB 1 intervention group's liver tissues showed a significantly higher degree of fibrosis (x2 =21.63,P =0),with more coarse fiber cords.The expression of ZEB1 and TGFβ1 was significantly higher in the model group than in the control group (both P ＜ 0.05).However,the ZEB 1 intervention group showed the highest levels of ZEB 1 and TGFβ1 expression (vs.model group,P ＜ 0.05).Conclusion ZEB 1 may promote the development of liver fibrosis in rats through a mechanism involving the TGFβ/Smad signaling pathway.