解整合素-金属蛋白酶9在小鼠急性酒精性肝损伤过程中的表达分析
|
摘要
目的研究解整合素-金属蛋白酶9(ADAM9)在小鼠急性酒精性肝损伤过程中的表达变化规律。方法将50只健康雄性小鼠随机分为正常组10只和实验组40只。实验组小鼠用56度乙醇12 m L·kg(-1)灌胃诱导小鼠急性肝损伤模型,分别于6,12,18和24 h后各取10只小鼠摘除眼球取血并分离血清,同时分离正常组血清,检测谷草转氨酶(AST)活性;颈椎脱臼处死小鼠并取肝,用蛋白印迹法测定正常组和实验组小鼠肝组织中ADAM9蛋白的表达变化。结果小鼠经乙醇灌胃后0,6,12,18,24 h的AST活性分别为(112±25),(316±31),(767±45),(1020±49),(525±23)U·L(-1),灌胃后6 h,血清AST活性显著上升(P<0.05),随着时间的增加,血清AST活性持续升高,18 h后达到最高值(P<0.01),然后随着肝损伤的修复,血清AST活性开始出现下降,24 h后酶活性仍高于正常组(P<0.05)。免疫印迹结果显示,乙醇灌胃后6 h,ADAM9的蛋白表达量显著上升(P<0.05),随着时间的增加,ADAM9的蛋白表达量持续升高,18 h后达到最高值(P<0.01),然后随着肝损伤的修复,ADAM9的蛋白表达量开始出现下降。结论 ADAM9在小鼠急性酒精性肝损伤过程中的先升高后降低且变化显著,ADAM9可能起到了促进酒精性肝损伤的作用。
Objective To investigate the expressed dynamic change of a disintegrin and metalloprotease 9( ADAM9) during acute liver injury induced by alcohol in mice. Methods Fifty healthy male mice were randomly divided into normal group( n = 10) and experimental group( n =40). The mice in experimental group were respectively drawn blood by removing the eyeballs and serum was separated to detect the activity of aspartate aminotransferase( AST) at 6,24,42 and 72 h after liver injury by alcohol( 12 mL·kg(-1)) with 56 ° alcohol,at the same time,the serum of normal group was separated,and the activity of AST was detected.The mice were sacrificed by cervical dislocation and the livers were taken,and the expression of ADAM9 protein in liver tissue of mice was determined by Western blot at different time points after the mice and alcohol were fed intragastrically. Results The AST at 0,6,12,18,24 h after administration with alcohol were( 112 ± 25),( 316 ± 31),( 767 ± 45),( 1020 ± 49),( 525 ± 23) U·L(-1),the activity of serum AST enzyme significantly increased at 6 h after alcohol( P < 0. 05).The AST activity of serum increased continuously with the increase of time and reached the highest value after 18 h( P < 0. 01),and then with liver damage repair,serum AST activity began to decline,the level at 24 h after enzyme activity was still higher than that in normal group( P < 0. 05). Western blot results showed that after 6 h of intragastric administration of alcohol,the expression of ADAM9 protein increased significantly( P < 0. 05). With the increase of time,the protein expression of ADAM9 continued to increase,reaching the highest value after 18 h( P < 0. 01),then with the repair of liver injury,ADAM9 Protein expression began to decline. Conclusion ADAM9 in mice with acute alcoholic liver injury increased first and then decreased and changed significantly,ADAM9 may play a role in promoting liver injury.
Objective To investigate the expressed dynamic change of a disintegrin and metalloprotease 9( ADAM9) during acute liver injury induced by alcohol in mice. Methods Fifty healthy male mice were randomly divided into normal group( n = 10) and experimental group( n =40). The mice in experimental group were respectively drawn blood by removing the eyeballs and serum was separated to detect the activity of aspartate aminotransferase( AST) at 6,24,42 and 72 h after liver injury by alcohol( 12 mL·kg(-1)) with 56 ° alcohol,at the same time,the serum of normal group was separated,and the activity of AST was detected.The mice were sacrificed by cervical dislocation and the livers were taken,and the expression of ADAM9 protein in liver tissue of mice was determined by Western blot at different time points after the mice and alcohol were fed intragastrically. Results The AST at 0,6,12,18,24 h after administration with alcohol were( 112 ± 25),( 316 ± 31),( 767 ± 45),( 1020 ± 49),( 525 ± 23) U·L(-1),the activity of serum AST enzyme significantly increased at 6 h after alcohol( P < 0. 05).The AST activity of serum increased continuously with the increase of time and reached the highest value after 18 h( P < 0. 01),and then with liver damage repair,serum AST activity began to decline,the level at 24 h after enzyme activity was still higher than that in normal group( P < 0. 05). Western blot results showed that after 6 h of intragastric administration of alcohol,the expression of ADAM9 protein increased significantly( P < 0. 05). With the increase of time,the protein expression of ADAM9 continued to increase,reaching the highest value after 18 h( P < 0. 01),then with the repair of liver injury,ADAM9 Protein expression began to decline. Conclusion ADAM9 in mice with acute alcoholic liver injury increased first and then decreased and changed significantly,ADAM9 may play a role in promoting liver injury.
引文
[1]BRUHA R,DVORAK K,PETRTYL J.Alcoholic liver disease[J].World J Hepatol,2012,4(3):81-90.
[2]厉有名,范建高,王炳元,等.酒精性肝病诊疗指南[J].临床肝胆病杂志,2010,4(3):229-232.
[3]SUH Y G,JEONG W I.Hepatic stellate cells and innate immunity in alcoholic liver disease[J].World J Gastroenterology,2011,17(20):2543-2551.
[4]DUFFY M J,MCKIERNAN E,O'DONOVAN N,et al.Role of ADAMs in cancer formation and progression[J].Clin Cancer Res,2009,15(4):1140-1144.
[5]MAROLDA R,CIOTTI M T,MATRONE C,et al.Substance P activates ADAM9 mRNA expression and inducesα-secretasemediated amyloid precursor protein cleavage[J].Neuropharmacology,2012,62(5-6):1954-1963.
[6]肖娟,张瑞芬,黄菲,等.两种不同的酒精摄入方式诱导的小鼠酒精性肝损伤模型比[J].中国比较医学杂志,2016,26(6):11-17.
[7]岳展伊,唐古生,杜大海,等.全自动生化分析仪检测酶活性过程中底物耗尽的监测和处理[J].国际检验医学杂志,2010,31(4):414-415.
[8]LI S Q,LI R F,XI S M,et al.Systematical analysis of impacts of heat stress on the proliferation,apoptosis and metabolism of mouse hepatocyte[J].J Physiol Sci,2012,62(1):29-43.
[9]RAYMOND E,FAIVRE S,ARMAND J P.Epidermal growth factor receptor tyrosine kinase as a target for anticancer therapy[J].Drugs,2000,60(Suppl.1):S15-S23.
[10]陈建明,庄时刚,刘军伟,等.ADAM9在胃癌中的表达及其临床意义[J].浙江医学,2011,33(7):1066-1068.
[11]SCHWETTMANN L,WEHMEIER M,JOKOVIC D,et al.Hepatic expression of A disintegrin and metalloproteinase(ADAM)and ADAMs with thrombospondin motives(ADAM-TS)enzymes in patients with chronic liver diseases[J].J Hepatol,2008,49(2):243-250.
[12]李三强,韩红梅,卢华杰,等.解整合素-金属蛋白酶9在对乙酰氨基酚诱导的小鼠急性肝损伤中的表达分析[J].中国临床药理学杂志,2015,31(12):1143-1145.
[2]厉有名,范建高,王炳元,等.酒精性肝病诊疗指南[J].临床肝胆病杂志,2010,4(3):229-232.
[3]SUH Y G,JEONG W I.Hepatic stellate cells and innate immunity in alcoholic liver disease[J].World J Gastroenterology,2011,17(20):2543-2551.
[4]DUFFY M J,MCKIERNAN E,O'DONOVAN N,et al.Role of ADAMs in cancer formation and progression[J].Clin Cancer Res,2009,15(4):1140-1144.
[5]MAROLDA R,CIOTTI M T,MATRONE C,et al.Substance P activates ADAM9 mRNA expression and inducesα-secretasemediated amyloid precursor protein cleavage[J].Neuropharmacology,2012,62(5-6):1954-1963.
[6]肖娟,张瑞芬,黄菲,等.两种不同的酒精摄入方式诱导的小鼠酒精性肝损伤模型比[J].中国比较医学杂志,2016,26(6):11-17.
[7]岳展伊,唐古生,杜大海,等.全自动生化分析仪检测酶活性过程中底物耗尽的监测和处理[J].国际检验医学杂志,2010,31(4):414-415.
[8]LI S Q,LI R F,XI S M,et al.Systematical analysis of impacts of heat stress on the proliferation,apoptosis and metabolism of mouse hepatocyte[J].J Physiol Sci,2012,62(1):29-43.
[9]RAYMOND E,FAIVRE S,ARMAND J P.Epidermal growth factor receptor tyrosine kinase as a target for anticancer therapy[J].Drugs,2000,60(Suppl.1):S15-S23.
[10]陈建明,庄时刚,刘军伟,等.ADAM9在胃癌中的表达及其临床意义[J].浙江医学,2011,33(7):1066-1068.
[11]SCHWETTMANN L,WEHMEIER M,JOKOVIC D,et al.Hepatic expression of A disintegrin and metalloproteinase(ADAM)and ADAMs with thrombospondin motives(ADAM-TS)enzymes in patients with chronic liver diseases[J].J Hepatol,2008,49(2):243-250.
[12]李三强,韩红梅,卢华杰,等.解整合素-金属蛋白酶9在对乙酰氨基酚诱导的小鼠急性肝损伤中的表达分析[J].中国临床药理学杂志,2015,31(12):1143-1145.