临床和实验医学杂志
臨床和實驗醫學雜誌
림상화실험의학잡지
JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE
2014年
17期
1393-1396
,共4页
何培元%王明娟%张聪%马立新%侯志平%李炳庆
何培元%王明娟%張聰%馬立新%侯誌平%李炳慶
하배원%왕명연%장총%마립신%후지평%리병경
大鼠模型%酒精性肝损伤%高脂饲料%骨形成蛋白 7%Smad6
大鼠模型%酒精性肝損傷%高脂飼料%骨形成蛋白 7%Smad6
대서모형%주정성간손상%고지사료%골형성단백 7%Smad6
Rat model%Alcoholic liver disease%High fat diet%BMP - 7%Smad6
目的:探讨 BMP -7/ Smad6信号传导通路在酒精性肝病发病机制中的作用并检测其在酒精性肝病大鼠模型中的基因水平和蛋白水平。方法以自由摄取白酒加高脂饲料诱导酒精性肝纤维化大鼠模型,并比较普通饲料与高脂饲料对酒精性肝病的影响。实验结束后取各实验组肝组织观察病理学变化,用生化检测仪检测大鼠血浆 ALT、AST变化。用实时 RCR 检测肝的骨形成蛋白7(BMP -7)和 Smad6 mRNA 水平变化,并用 Western Blot 检测蛋白水平变化。结果肝脏组织形态学改变显示正常饲料组肝组织出现脂肪肝表现,高脂饲料组脂肪变性程度加重并出现了肝炎及胶原纤维增生,而对照组大鼠肝脏组织结构正常。生化分析仪检测 ALT、AST 结果显示:至实验结束时,实验组比对照组转氨酶活性明显升高( P ﹤0.05)。BMP 和 Smad6这两个基因的蛋白丰度与它们各自的 mRNA 表达水平呈正相关,与对照组比较两个实验组 BMP -7表达水平降低,并与肝脏病理损伤程度呈负相关,两个实验组 Smad6表达水平较对照组升高,但与肝脏病理变化严重程度无相关性。结论饮用白酒水溶液结合高脂饲料的方法可以成功建立酒精性肝损伤的大鼠模型,BMP -7/ Smad6信号通路的失活促进了肝纤维化的形成。
目的:探討 BMP -7/ Smad6信號傳導通路在酒精性肝病髮病機製中的作用併檢測其在酒精性肝病大鼠模型中的基因水平和蛋白水平。方法以自由攝取白酒加高脂飼料誘導酒精性肝纖維化大鼠模型,併比較普通飼料與高脂飼料對酒精性肝病的影響。實驗結束後取各實驗組肝組織觀察病理學變化,用生化檢測儀檢測大鼠血漿 ALT、AST變化。用實時 RCR 檢測肝的骨形成蛋白7(BMP -7)和 Smad6 mRNA 水平變化,併用 Western Blot 檢測蛋白水平變化。結果肝髒組織形態學改變顯示正常飼料組肝組織齣現脂肪肝錶現,高脂飼料組脂肪變性程度加重併齣現瞭肝炎及膠原纖維增生,而對照組大鼠肝髒組織結構正常。生化分析儀檢測 ALT、AST 結果顯示:至實驗結束時,實驗組比對照組轉氨酶活性明顯升高( P ﹤0.05)。BMP 和 Smad6這兩箇基因的蛋白豐度與它們各自的 mRNA 錶達水平呈正相關,與對照組比較兩箇實驗組 BMP -7錶達水平降低,併與肝髒病理損傷程度呈負相關,兩箇實驗組 Smad6錶達水平較對照組升高,但與肝髒病理變化嚴重程度無相關性。結論飲用白酒水溶液結閤高脂飼料的方法可以成功建立酒精性肝損傷的大鼠模型,BMP -7/ Smad6信號通路的失活促進瞭肝纖維化的形成。
목적:탐토 BMP -7/ Smad6신호전도통로재주정성간병발병궤제중적작용병검측기재주정성간병대서모형중적기인수평화단백수평。방법이자유섭취백주가고지사료유도주정성간섬유화대서모형,병비교보통사료여고지사료대주정성간병적영향。실험결속후취각실험조간조직관찰병이학변화,용생화검측의검측대서혈장 ALT、AST변화。용실시 RCR 검측간적골형성단백7(BMP -7)화 Smad6 mRNA 수평변화,병용 Western Blot 검측단백수평변화。결과간장조직형태학개변현시정상사료조간조직출현지방간표현,고지사료조지방변성정도가중병출현료간염급효원섬유증생,이대조조대서간장조직결구정상。생화분석의검측 ALT、AST 결과현시:지실험결속시,실험조비대조조전안매활성명현승고( P ﹤0.05)。BMP 화 Smad6저량개기인적단백봉도여타문각자적 mRNA 표체수평정정상관,여대조조비교량개실험조 BMP -7표체수평강저,병여간장병리손상정도정부상관,량개실험조 Smad6표체수평교대조조승고,단여간장병리변화엄중정도무상관성。결론음용백주수용액결합고지사료적방법가이성공건립주정성간손상적대서모형,BMP -7/ Smad6신호통로적실활촉진료간섬유화적형성。
Objective To investigate the function of BMP - 7 / Smad6 signaling pathway in alcoholic liver disease rat model and detect the two genes in both mRNA and protein levels. Methods Rats were fed with alcohol and high fat diet to establish alcoholic liver disease model. In the end of the experiment,liver of rats were dissected for histological studies whereas plasma obtained from rats was taken for biochemical testing to detect the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST). mRNA was extracted from the liver to perform real- time PCR in order to detect the mRNA level of bone morphogenetic proteins 7(BMP - 7)and Smad6. Western blot was performed to determine the protein level of BMP - 7 and Smad6. Results Livers from the experimental group fed with alcohol demonstrated histopathological changes such as fatty change,inflammation as well as fibrosis;however,no pathological changes were observed in the livers of rats in normal control group. Bio-chemical testing results showed that levels of AST and ALT were higher in experimental group compared to normal control group. The results of PCR and western blot showed that the mRNA levels of BMP - 7 and Smad6 positively correlated to the corresponding protein levels. BMP - 7 level of normal control group was higher than that of experimental group,and the level was negatively correlated to the severity of hepatic pathological changes of liver. Smad6 levels were higher in experimental group compared to the normal control group,however,there was no correlation between the level of Smad6 and hepatic pathological changes. Conclusion Feeding high fat diet with alcohol dilution on rats is a convenient,low cost method in establishing animal model of alcoholic liver disease. Inactivation of BMP - 7 / Smad6 pathway contributes to the development of alcoholic liver fibrosis.