中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2009年
5期
317-321
,共5页
潘浩%窦爱霞%陈尉华%周琨%陈婷%朱长清%归茜%房静远%曾民德%陆伦根
潘浩%竇愛霞%陳尉華%週琨%陳婷%硃長清%歸茜%房靜遠%曾民德%陸倫根
반호%두애하%진위화%주곤%진정%주장청%귀천%방정원%증민덕%륙륜근
脂肪肝%转化生长因子β1%Smad4蛋白质%疾病模型,动物
脂肪肝%轉化生長因子β1%Smad4蛋白質%疾病模型,動物
지방간%전화생장인자β1%Smad4단백질%질병모형,동물
Fatty liver%Transforming growth factor betal%Smad4 protein%Disease model,animal
目的 观察转化生长因子(TGF)β1、转化生长因子β受体(TβR)-Ⅰ、TβR-Ⅲ和胞内Smad4信号通道蛋白以及转录因子激活剂蛋白-1(AP-1)家族中Jun蛋白质因子在大鼠非酒精性脂肪性肝病(NAFLD)中的表达.探讨NAFLD发生肝纤维化的可能机制.方法 将18只大鼠分为对照组与模型组,每组9只.对照组喂饲普通饲料,模型组喂饲高脂饲料(88%标准普通饲料+10%猪油+2%胆固醇)造模.在20周处死所有大鼠,免疫组化法检测TGFβ1、Smad4的表达;RT-PCR检测肝组织TGFβ1、TβR-Ⅰ、TβR-Ⅱ的表达;Western印迹法测定磷酸化C-Jun蛋白的表达.结果 成功建立NAFLD动物模型,免疫组化法检测结果显示,TGFβ1和Smad4在对照组大鼠肝实质细胞的胞质内均有少量表达.TGFβ1在模型组大鼠肝实质细胞胞质、肝血窦周围的表达明显增强,尤其在汇管区更为明显.Smad4在模型组大鼠肝实质细胞胞质表达亦明显增强,而在汇管区的表达更为明显.RT-PCR检测结果显示,模型组大鼠肝组织TGFβ1、TβR-Ⅰ、TβR-ⅡmRNA的A值分别为0.46±0.12、5.24±2.70和3.35±1.95,明显高于对照组(0.21±0.09、1.36±0.77和0.52±0.19,P值均＜0.01).Western印迹检测显示,模型组磷酸化C-Jun蛋白表达同内参灰度值比为0.93±0.41,较对照组显著增高(0.32±0.25,P=0.001).结论 TGFβ1/Smad4信号通路可能参与了NAFLD进展为肝纤维化的过程.阻断TGFβ/Smad4信号传导通路,可开辟治疗NAFLD新的思路.
目的 觀察轉化生長因子(TGF)β1、轉化生長因子β受體(TβR)-Ⅰ、TβR-Ⅲ和胞內Smad4信號通道蛋白以及轉錄因子激活劑蛋白-1(AP-1)傢族中Jun蛋白質因子在大鼠非酒精性脂肪性肝病(NAFLD)中的錶達.探討NAFLD髮生肝纖維化的可能機製.方法 將18隻大鼠分為對照組與模型組,每組9隻.對照組餵飼普通飼料,模型組餵飼高脂飼料(88%標準普通飼料+10%豬油+2%膽固醇)造模.在20週處死所有大鼠,免疫組化法檢測TGFβ1、Smad4的錶達;RT-PCR檢測肝組織TGFβ1、TβR-Ⅰ、TβR-Ⅱ的錶達;Western印跡法測定燐痠化C-Jun蛋白的錶達.結果 成功建立NAFLD動物模型,免疫組化法檢測結果顯示,TGFβ1和Smad4在對照組大鼠肝實質細胞的胞質內均有少量錶達.TGFβ1在模型組大鼠肝實質細胞胞質、肝血竇週圍的錶達明顯增彊,尤其在彙管區更為明顯.Smad4在模型組大鼠肝實質細胞胞質錶達亦明顯增彊,而在彙管區的錶達更為明顯.RT-PCR檢測結果顯示,模型組大鼠肝組織TGFβ1、TβR-Ⅰ、TβR-ⅡmRNA的A值分彆為0.46±0.12、5.24±2.70和3.35±1.95,明顯高于對照組(0.21±0.09、1.36±0.77和0.52±0.19,P值均＜0.01).Western印跡檢測顯示,模型組燐痠化C-Jun蛋白錶達同內參灰度值比為0.93±0.41,較對照組顯著增高(0.32±0.25,P=0.001).結論 TGFβ1/Smad4信號通路可能參與瞭NAFLD進展為肝纖維化的過程.阻斷TGFβ/Smad4信號傳導通路,可開闢治療NAFLD新的思路.
목적 관찰전화생장인자(TGF)β1、전화생장인자β수체(TβR)-Ⅰ、TβR-Ⅲ화포내Smad4신호통도단백이급전록인자격활제단백-1(AP-1)가족중Jun단백질인자재대서비주정성지방성간병(NAFLD)중적표체.탐토NAFLD발생간섬유화적가능궤제.방법 장18지대서분위대조조여모형조,매조9지.대조조위사보통사료,모형조위사고지사료(88%표준보통사료+10%저유+2%담고순)조모.재20주처사소유대서,면역조화법검측TGFβ1、Smad4적표체;RT-PCR검측간조직TGFβ1、TβR-Ⅰ、TβR-Ⅱ적표체;Western인적법측정린산화C-Jun단백적표체.결과 성공건립NAFLD동물모형,면역조화법검측결과현시,TGFβ1화Smad4재대조조대서간실질세포적포질내균유소량표체.TGFβ1재모형조대서간실질세포포질、간혈두주위적표체명현증강,우기재회관구경위명현.Smad4재모형조대서간실질세포포질표체역명현증강,이재회관구적표체경위명현.RT-PCR검측결과현시,모형조대서간조직TGFβ1、TβR-Ⅰ、TβR-ⅡmRNA적A치분별위0.46±0.12、5.24±2.70화3.35±1.95,명현고우대조조(0.21±0.09、1.36±0.77화0.52±0.19,P치균＜0.01).Western인적검측현시,모형조린산화C-Jun단백표체동내삼회도치비위0.93±0.41,교대조조현저증고(0.32±0.25,P=0.001).결론 TGFβ1/Smad4신호통로가능삼여료NAFLD진전위간섬유화적과정.조단TGFβ/Smad4신호전도통로,가개벽치료NAFLD신적사로.
Objective To investigate the expression of transforming growth factor β1,transforming growth factor beta receptor(TBR)Ⅰ,TβR Ⅱ,Smad4 and C-Jun in rats with nonalcoholic fatty liver disease(NAFLD)and to find out the mechanisms of liver fibrosis in patients with NAFLD.Methods A total of 18 male SD rats were randomly divided into normal control group(n=9)and model group(n=9).The rats in control group were fed with normal diet,and those in model group were fed with fat-rich diet(consisted of 10%lard oil+2%cholesterol).An rats were sacrificed at the 20th week.The levels of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were examined by RT-PCR.The expressions of TGFβ1 and Smad4 in liver tissue were detected by immunohistochemistry.The expression of C-Jun protein was detected by Western blotting.Results The NAFLD model was successfully established.The immunohistochemistry examination revealed that TGFβ1 and Smad4 were expressed weekly in control group,but strongly expressed in model group.RT-PCR showed that A values of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were 0.46±0.12,5.z4±2.70 and 3.35±1.95,respectively,in model group,which were higher than those in control group(0.21±0.09,1.36±0.77 and 0.52±0.19,all P values＜0.01).The Western blotting results demonstrated that the expression of C-Jun protein in model group(0.93±0.41)was higher than that in control group (0.32±0.25,P=0.001).Conclusion TGFβ1/Smad4 signal pathway might be involved in the development of hepatic fibrosis in NAFLD.Blocking TGFβ1/Smad4 signal pathway will be helpful in treatment of NAFLD.
