重组人MG53对CCl_4诱导的急性肝损伤的保护作用
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摘要
目的研究重组人MG53(rh MG53)对四氯化碳(CCl_4)诱导的急性肝损伤的保护作用及其相关机制。方法 40只雄性C57小鼠采用完全随机化分组法分成4组:正常对照组、rh MG53组、CCl_4组、rh MG53+CCl_4组,每组10只。CCl_4组、rh MG53+CCl_4组小鼠予0.1%CCl_4花生油溶液2 m L/kg腹腔内注射,rh MG53+CCl_4组在注射CCl_4前30 min予rh MG53 1 mg/kg肌肉注射。rh MG53组予rh MG53 1 mg/kg肌肉注射。正常进食进水24 h后眼球取血分离血清,测定谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)活性。取小鼠肝脏,观察肝脏的组织学改变。以体外培养的人肝癌细胞(Hep G2)为靶细胞,观察rh MG53对CCl_4诱导的细胞损伤的保护作用。结果 rh MG53降低小鼠血清ALT、AST、LDH水平,病理镜检显示rh MG53可改善肝脏的病理结构,差异均有统计学意义(P<0.05);rh MG53可以修复Hep G2细胞膜,保护CCl_4对Hep G2细胞的损伤。结论 rh MG53改善CCl_4诱导的急性肝细胞损伤,保护肝脏功能。
Objective To determine the protective effect of recombinant human MG53( rh MG53) on acute liver injury in mice induced by carbon tetrachloride( CCl_4),and investigate its underlying mechanism.Methods Male C57 mice( n = 40) were randomly divided into 4 groups( n = 10),that is,normal control group,rh MG53 group,CCl_4 group,and rh MG53 + CCl_4 group. The animals of the CCl_4 group and rh MG53 +CCl_4 group were injected intraperitoneally with 0. 1% CCl_4 peanut oil solution at a dose of 2 m L/kg,and the mice of the latter group were injected intramuscularly with 1 mg/kg rh MG53 in 0. 5 h before CCl_4 injection. In24 h after normal feeding,blood samples were collected after eyeball extirpating,and the serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST) and lactate dehydrogenase( LDH) were measured. The liver tissues of the mice were also harvested for histological changes of the livers. Human hepatocellular carcinoma cells( Hep G2) were used as target cells to observe the protective effect of rh MG53 on CCl_4-induced cell injury. Results Rh MG53 treatment reduced serum ALT,AST and LDH levels,and significant attenuated the pathological injuries induced by CCl_4( P < 0. 05). rh MG53 repaired cell membrane and reduced CCl_4-induced damage to Hep G2 cells. Conclusion rh MG53 shows protective effect on CCl_4-induced acute hepatocellular injury and protects the liver function in mice.
Objective To determine the protective effect of recombinant human MG53( rh MG53) on acute liver injury in mice induced by carbon tetrachloride( CCl_4),and investigate its underlying mechanism.Methods Male C57 mice( n = 40) were randomly divided into 4 groups( n = 10),that is,normal control group,rh MG53 group,CCl_4 group,and rh MG53 + CCl_4 group. The animals of the CCl_4 group and rh MG53 +CCl_4 group were injected intraperitoneally with 0. 1% CCl_4 peanut oil solution at a dose of 2 m L/kg,and the mice of the latter group were injected intramuscularly with 1 mg/kg rh MG53 in 0. 5 h before CCl_4 injection. In24 h after normal feeding,blood samples were collected after eyeball extirpating,and the serum levels of alanine aminotransferase( ALT),aspartate aminotransferase( AST) and lactate dehydrogenase( LDH) were measured. The liver tissues of the mice were also harvested for histological changes of the livers. Human hepatocellular carcinoma cells( Hep G2) were used as target cells to observe the protective effect of rh MG53 on CCl_4-induced cell injury. Results Rh MG53 treatment reduced serum ALT,AST and LDH levels,and significant attenuated the pathological injuries induced by CCl_4( P < 0. 05). rh MG53 repaired cell membrane and reduced CCl_4-induced damage to Hep G2 cells. Conclusion rh MG53 shows protective effect on CCl_4-induced acute hepatocellular injury and protects the liver function in mice.
引文
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[2]OH I S,PARK S H.Immune-mediated liver injury in hepatitis B virus infection[J].Immune Netw,2015,15(4):191-198.DOI:10.4110/in.2015.15.4.191.
[3]WEISLEDER N,TAKIZAWA N,LIN P,et al.Recombinant MG53 protein modulates therapeutic cell membrane repair in treatment of muscular dystrophy[J].Sci Transl Med,2012,4(139):139ra85.DOI:10.1126/scitranslmed.3003921.
[4]CAI C,MASUMIYA H,WEISLEDER N,et al.MG53 nucleates assembly of cell membrane repair machinery[J].Nat Cell Biol,2009,11(1):56-64.DOI:10.1038/ncb1812.
[5]ZHU H,HOU J,ROE J L,et al.Amelioration of ischemiareperfusion-induced muscle injury by the recombinant human MG53 protein[J].Muscle Nerve,2015,52(5):852-858.DOI:10.1002/mus.24619.
[6]WANG X,XIE W,ZHANG Y,et al.Cardioprotection of ischemia/reperfusion injury by cholesterol-dependent MG53-mediated membrane repair[J].Circ Res,2010,107(1):76-83.DOI:10.1161/CIRCRESAHA.109.215822.
[7]CAO C M,ZHANG Y,WEISLEDER N,et al.MG53 constitutes a primary determinant of cardiac ischemic preconditioning[J].Circulation,2010,121(23):2565-2574.DOI:10.1161/CIRCULATIONAHA.110.954628.
[8]LIU J,ZHU H,ZHENG Y,et al.Cardioprotection of recombinant human MG53 protein in a porcine model of ischemia and reperfusion injury[J].J Mol Cell Cardiol,2015,80:10-19.DOI:10.1016/j.yjmcc.2014.12.010.
[9]JIA Y,CHEN K,LIN P,et al.Treatment of acute lung injury by targeting MG53-mediated cell membrane repair[J].Nat Commun,2014,5:4387.DOI:10.1038/ncomms5387.
[10]DUANN P,LI H,LIN P,et al.MG53-mediated cell membrane repair protects against acute kidney injury[J].Sci Transl Med,2015,7(279):279ra36.DOI:10.1126/scitranslmed.3010755.
[11]YAO Y G,ZHANG B,ZHU H,et al.MG53 permeates through blood-brain barrier to protect ischemic brain injury[J].Oncotarget,2016,7(16):22474-22485.DOI:10.18632/oncotarget.7965.
[12]CUBERO F J,ZOUBEK M E,HU W,et al.Combined activities of JNK1 and JNK2 in hepatocytes protect against toxic liver injury[J].Gastroenterology,2016,150(4):968-981.DOI:10.1053/j.gastro.2015.12.019.
[13]WEBER L W,BOLL M,STAMPFL A.Hepatotoxicity and mechanism of action of haloalkanes:carbon tetrachloride as a toxicological model[J].Crit Rev Toxicol,2003,33(2):105-136.DOI:10.1080/713611034.
[14]ZHANG Y,WU H K,LV F,et al.MG53:biological function and potential as a therapeutic target[J].Mol Pharmacol,2017,92(3):211-218.DOI:10.1124/mol.117.108241.
[15]LI H,DUANN P,LIN P H,et al.Modulation of wound healing and scar formation by MG53 protein-mediated cell membrane repair[J].J Biol Chem,2015,290(40):24592-24603.DOI:10.1074/jbc.M115.680074.
[16]YAO W,LI H,HAN X,et al.MG53 anchored by dysferlin to cell membrane reduces hepatocyte apoptosis which induced by ischaemia/reperfusion injury in vivo and in vitro[J].J Cell Mol Med,2017.[Epub ahead of print].DOI:10.1111/jcmm.13171.