中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2014年
2期
89-93
,共5页
王丽萍%董进中%曹肃婷%张赛男%林镯%王晓东%陈永平
王麗萍%董進中%曹肅婷%張賽男%林鐲%王曉東%陳永平
왕려평%동진중%조숙정%장새남%림탁%왕효동%진영평
肝硬化%骨形态发生蛋白质类%受体,表皮生长因子%转化生长因子β1%疾病模型,动物
肝硬化%骨形態髮生蛋白質類%受體,錶皮生長因子%轉化生長因子β1%疾病模型,動物
간경화%골형태발생단백질류%수체,표피생장인자%전화생장인자β1%질병모형,동물
Liver cirrhosis%Bone morphogenetic proteins%Receptor,epidermal growth factor%Transforming growth factor beta1%Disease models,animal
目的 观察肝纤维化小鼠模型中表皮生长因子受体(EGFR)的动态表达,以及骨形成蛋白-7(BMP-7)对其表达的影响,探讨治疗纤维化的新靶点.方法 30只健康雄性ICR小鼠随机数字表法分为正常对照组(6只)、肝纤维化模型组(18只,包括4、8、12周3个亚组)和BMP-7干预组(6只),皮下注射四氯化碳制备肝纤维化小鼠模型,BMP-7干预组注射四氯化碳8周后腹腔注射人重组BMP-7,持续4周.采用HE和Manson染色观察肝组织病理变化,并进行肝纤维化半定量评分;下腔静脉采血检测ALT、AST和白蛋白;采用实时荧光定量PCR和Western印迹法分别检测各组TGF-β1 mRNA和TGF-β1、EGFR、pEGFR蛋白表达情况.计量资料比较采用方差分析,相关性采用Pearson相关分析.结果 成功建立小鼠肝纤维化模型,肝纤维化模型组血清ALT、AST均逐渐升高,12周时最高,白蛋白逐渐降低,12周时最低,BMP-7干预组各指标均有所缓解[ALT:(153.9±18.1) U/L比(191.3±24.5) U/L;AST:(177.8±19.2) U/L比(206.6±25.0)U/L;白蛋白:(25.4±0.9)g/L比(22.2±1.2) g/L;均P＜0.05].肝纤维化组随着纤维化的进展,TGF-β1、EGFR及pEGFR的蛋白表达量均逐渐升高,于12周时达到高峰,BMP-7干预后,上述3种蛋白的表达水平均明显降低(TGF-β1:0.256±0.006比0.287±0.014;EGFR:1.061±0.017比1.094±0.014; pEGFR:0.855±0.053比1.007±0.063;均P＜0.05).直线相关分析显示,TGF-β1与EGFR及pEGFR表达量均呈正相关(rs值分别为0.895、0.859,均P＜0.05).结论 BMP-7能改善小鼠肝纤维化,其作用机制可能是调节EGFR和TGF-β1的表达.
目的 觀察肝纖維化小鼠模型中錶皮生長因子受體(EGFR)的動態錶達,以及骨形成蛋白-7(BMP-7)對其錶達的影響,探討治療纖維化的新靶點.方法 30隻健康雄性ICR小鼠隨機數字錶法分為正常對照組(6隻)、肝纖維化模型組(18隻,包括4、8、12週3箇亞組)和BMP-7榦預組(6隻),皮下註射四氯化碳製備肝纖維化小鼠模型,BMP-7榦預組註射四氯化碳8週後腹腔註射人重組BMP-7,持續4週.採用HE和Manson染色觀察肝組織病理變化,併進行肝纖維化半定量評分;下腔靜脈採血檢測ALT、AST和白蛋白;採用實時熒光定量PCR和Western印跡法分彆檢測各組TGF-β1 mRNA和TGF-β1、EGFR、pEGFR蛋白錶達情況.計量資料比較採用方差分析,相關性採用Pearson相關分析.結果 成功建立小鼠肝纖維化模型,肝纖維化模型組血清ALT、AST均逐漸升高,12週時最高,白蛋白逐漸降低,12週時最低,BMP-7榦預組各指標均有所緩解[ALT:(153.9±18.1) U/L比(191.3±24.5) U/L;AST:(177.8±19.2) U/L比(206.6±25.0)U/L;白蛋白:(25.4±0.9)g/L比(22.2±1.2) g/L;均P＜0.05].肝纖維化組隨著纖維化的進展,TGF-β1、EGFR及pEGFR的蛋白錶達量均逐漸升高,于12週時達到高峰,BMP-7榦預後,上述3種蛋白的錶達水平均明顯降低(TGF-β1:0.256±0.006比0.287±0.014;EGFR:1.061±0.017比1.094±0.014; pEGFR:0.855±0.053比1.007±0.063;均P＜0.05).直線相關分析顯示,TGF-β1與EGFR及pEGFR錶達量均呈正相關(rs值分彆為0.895、0.859,均P＜0.05).結論 BMP-7能改善小鼠肝纖維化,其作用機製可能是調節EGFR和TGF-β1的錶達.
목적 관찰간섬유화소서모형중표피생장인자수체(EGFR)적동태표체,이급골형성단백-7(BMP-7)대기표체적영향,탐토치료섬유화적신파점.방법 30지건강웅성ICR소서수궤수자표법분위정상대조조(6지)、간섬유화모형조(18지,포괄4、8、12주3개아조)화BMP-7간예조(6지),피하주사사록화탄제비간섬유화소서모형,BMP-7간예조주사사록화탄8주후복강주사인중조BMP-7,지속4주.채용HE화Manson염색관찰간조직병리변화,병진행간섬유화반정량평분;하강정맥채혈검측ALT、AST화백단백;채용실시형광정량PCR화Western인적법분별검측각조TGF-β1 mRNA화TGF-β1、EGFR、pEGFR단백표체정황.계량자료비교채용방차분석,상관성채용Pearson상관분석.결과 성공건립소서간섬유화모형,간섬유화모형조혈청ALT、AST균축점승고,12주시최고,백단백축점강저,12주시최저,BMP-7간예조각지표균유소완해[ALT:(153.9±18.1) U/L비(191.3±24.5) U/L;AST:(177.8±19.2) U/L비(206.6±25.0)U/L;백단백:(25.4±0.9)g/L비(22.2±1.2) g/L;균P＜0.05].간섬유화조수착섬유화적진전,TGF-β1、EGFR급pEGFR적단백표체량균축점승고,우12주시체도고봉,BMP-7간예후,상술3충단백적표체수평균명현강저(TGF-β1:0.256±0.006비0.287±0.014;EGFR:1.061±0.017비1.094±0.014; pEGFR:0.855±0.053비1.007±0.063;균P＜0.05).직선상관분석현시,TGF-β1여EGFR급pEGFR표체량균정정상관(rs치분별위0.895、0.859,균P＜0.05).결론 BMP-7능개선소서간섬유화,기작용궤제가능시조절EGFR화TGF-β1적표체.
Objective To investigate the dynamic expressions of epidermal growth factor receptor (EGFR) in mice with liver fibrosis and the effect of bone morphogenetic protein-7 (BMP-7) intervention on the expression of EGFR,and to explore a new therapy target for fibrosis.Methods A total of 30 healthy male ICR mice were randomly divided into three groups:6 mice in control group,18 mice in hepatic fibrosis group and 6 mice in BMP-7 intervention group.The model of mice with liver fibrosis was established by subcutaneous injection of carbon tetrachloride (CCl4) for 12 weeks.After administration of CCl4 for 8 weeks,human recombinant BMP-7 was given into mice in intervention group by intraperitoneal injection for 4 weeks.Hematoxylin-Eosin and Masson staining of liver tissues were employed to observe the pathological changes,and the semi-quantitative analysis of liver fibrosis was performed.Blood withdrawn from inferior vena cava was detected for levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST) and albumin (Alb).The expressions of transforming growth factor-β1 (TGF-β1)mRNA and TGF-β1,EGFR,phosphorylation EGFR (pEGFR) protein in each group were detected using quantitative real time polymerase chain reaction and Western blot.Measurement date was compared using analysis of variance and Pearson correlation analysis.Results The model of mice with liver fibrosis was successfully established.In model group,the serum levels of ALT and AST increased,while the level of Alb decreased gradually.All these biochemical index improved after intervention of BMP-7 (ALT:[153.9±18.1] U/L vs [191.3±24.5] U/L;AST:[177.8±19.2] U/L vs [206.6±25.0] U/L;Alb:[25.4±0.9] g/L vs [22.2±1.2] g/L; all P＜0.05).With the progress of fibrosis,TGF-β1,EGFR and pEGFR protein expressions increased gradually in model group and reached a peak at week 12,which was significantly different compared to the control group (all P＜0.05).In BMP-7 intervention group,the expressions of the three proteins decreased significantly compared to model group (TGF-β1:0.256 ± 0.006 vs 0.287±0.014,EGFR:1.061±0.017 vs 1.094±0.014,pEGFR:0.855±0.053 vs 1.007±0.063;all P＜0.05).Additionally,linear correlation analysis showed that expressions of both EGFR and pEGFR proteins were positively correlated with TGF-β1 protein (rs =0.895 and 0.859,respectively; both P＜0.05).Conclusions BMP-7 can suppress the pathogenesis of mouse liver fibrosis.The mechanism may rely on the regulation of EGFR and TGF-β1 expressions.
