胆道闭锁幼鼠肝脏组织中LOXL-2与MMP-7、MMP-9、TGF-β1的关系研究

Research on the relationship between LOXL-2 and MMP-7, MMP-9 and TGF-β1 in liver tissues of newborn mice with biliary atresia

目的研究探索LOXL-2在胆道闭锁(BA)病理发展中的作用,探讨LOXL-2与MMP-7、MMP-9、TGF-β1的关系。方法BALB/c新生幼鼠60只,随机分为对照组(n=30)和模型组(n=30),模型组腹腔注射恒河猴轮状病毒(RRV)建立幼鼠BA模型。分别于造模后7 d、15 d、28 d处死各组小鼠,以qPCR荧光定量检测肝脏组织中LOXL-2、MMP-7、MMP-9及TGF-β1的mRNA水平,ELISA检测LOXL-2、MMP-7、MMP-9及TGF-β1肝脏组织中蛋白表达水平。结果造模后15 d、28 d,模型组肝脏LOXL-2 mRNA、LOXL-2表达均高于对照组(P<0.05);模型组肝脏MMP-7 mRNA、MMP-7表达均高于对照组,到28 d时达到最高,差异均有统计学意义(P<0.05)。造模后15 d,模型组肝脏MMP-9 mRNA、TGF-β1 mRNA、MMP-9、TGF-β1表达高于对照组(P<0.05);造模后28 d,模型组肝脏MMP-9 mRNA、MMP-9表达与对照组无统计学差异(P>0.05),而TGF-β1 mRNA、TGF-β1表达持续升高且差异均有统计学意义(P<0.05)。LOXL-2的表达与TGF-β1、MMP-7的表达呈正相关(r=0.707,P<0.001;r=0.556,P=0.001),与MMP-9的表达无相关性(r=-0.022,P=0.907)。结论LOXL-2参与了胆道闭锁肝纤维化的病理过程,可能与TGF-β1、MMP-7协同促使肝纤维化。

Objective To investigate the role of LOXL-2 in the pathological development of biliary atresia(BA)and to explore the interaction among LOXL-2,MMP-7,MMP-9 and TGF-β1.Methods 60 Balb/c newborn mice were randomly divided into model group and control group(n=30).Model group was intraperitoneally injected with rhesus rotavirus(RRV)to establish BA model in mouse.Mice in each group were sacrificed at 7d,15d and 28d after model establishment respectively.The mRNA levels of LOXL-2,MMP-7,MMP-9 and TGF-β1 in liver tissues were detected by qPCR fluorescence,and LOXL-2,MMP-7,MMP-9 and TGF-β1 protein expression levels were detected by ELISA kit.Results The expressions of LOXL-2 mRNA and LOXL-2 level in the model group were significantly higher than those in the control group at 15 and 28 days after model establishment(P<0.05).The expressions of MMP-7 mRNA and MMP-7 in the liver tissue of model group were significantly higher than those of control group and reached the highest level at 28 days(P<0.05).The expressions of MMP-9 mRNA,TGF-β1 mRNA,MMP-9 and TGF-β1 in the liver tissue of model group were significantly higher than those of control group at 15 days after modeling(P<0.05),and there was no significant difference in the expressions of MMP-9 mRNA and MMP-9 at 28 days after modeling(P>0.05),while the expressions of TGF-β1 mRNA and TGF-β1 were continuously increased(P<0.05).The expressions of LOXL-2 were positively correlated with the expressions of TGF-β1 and MMP-7(r=0.707,P<0.001 and r=0.556,P=0.001),and had no correlation with the expressions of MMP-9(r=-0.022,P=0.907).Conclusion LOXL-2 is involved in the pathological process of hepatic fibrosis caused by biliary atresia,and may be synergistic with TGF-β1 and MMP-7 to promote liver fibrosis.