D-氨基半乳糖诱导的非人灵长类动物急性肝衰竭模型的建立
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摘要
目的探讨不同剂量D-氨基半乳糖(D-Gal)对非人灵长类动物食蟹猴的影响并建立不同程度急性肝衰竭(ALF)猴模型。方法将12只食蟹猴分为3组,每组4只,分别为低剂量组、中剂量组和高剂量组,给药剂量分别为0.23、0.25、0.27 g/kg。各组动物均在清醒状态(无麻醉)下,将相应剂量的D-Gal溶液经前臂静脉一次性注射入猴体内。记录猴的存活时间,观察是否出现消化道症状和肝性脑病症状。分别于给药前0 h,给药后12、24、36、48、60、72、96、120、144 h测量生命体征,留取血标本检测丙氨酸转氨酶(ALT)、总胆红素(TB)、凝血酶原时间(PT)、血氨等指标。取肝脏组织行苏木素-伊红(HE)染色观察病理学改变。结果低剂量组猴全部存活,仅出现一过性肝损伤,无肝性脑病症状,给药后60 h,肝功能和凝血指标达峰值,后逐渐恢复,120 h后基本恢复正常;中剂量组猴病程进展稍缓,在出现严重肝功能损伤及肝性脑病症状后逐渐恢复,仅1只死亡;高剂量组猴均出现肝性脑病症状后死亡,存活时间(72±13)h,肝功能严重受损。肝组织病理学检查示,低剂量组肝组织散在肝细胞坏死,炎症细胞浸润;中、高剂量组可见肝小叶结构不清,肝细胞片状坏死伴有明显出血。结论中、高剂量组均符合ALF模型标准,中剂量组ALF程度较轻,有利于肝移植手术等实验的开展;高剂量组ALF程度较重,适用于相关治疗方案疗效评价。
Objective To evaluate the effect of different doses of D-galactosamine(D-Gal) on non-human primate cynomolgus monkey and to establish a monkey model with different degree of acute liver failure(ALF). Methods Twelve cynomolgus monkeys were evenly divided into the low-, medium-and high-dose groups(n=4) with a dosage of 0.23, 0.25 and 0.27 g/kg, respectively. In each group, the corresponding dose of D-Gal solution was injected into the monkeys through the forearm vein at one time in a sober state(without anesthesia). The survival time of the cynomolgus monkeys was recorded. Digestive tract and hepatic encephalopathy symptoms were observed. Vital signs were measured at 0 h before and 12, 24, 36, 48, 60, 72, 96, 120 and 144 h after D-Gal administration. Alanine transaminase(ALT), total bilirubin(TB), prothrombin time(PT), blood ammonia and other parameters were detected from the blood samples. The liver tissues were prepared for hematoxylin-eosin(HE) staining to observe the pathological changes. Results All cynomolgus monkeys in the low-dose group survived and transient liver injury was noted without the hepatic encephalopathy symptoms. At 60 h after D-Gal administration, the liver function and coagulation indexes reached the peak, gradually recovered and then basically returned to the normal range at 120 h. In the medium-dose group, the course of disease was relatively slow and gradually recovered after the appearance of severe liver damage and hepatic encephalopathy symptoms and only one animal died. All cynomolgus monkeys in the high-dose group died after developing hepatic encephalopathy symptoms and severe liver damage with a mean survival time of(72±13) h. Pathological examination of liver tissue demonstrated that scattered liver cell necrosis and inflammatory cell infiltration were observed in the liver tissues of the low-dose group. In the medium-and high-dose groups, the hepatic lobule structure was not clear, and the liver cell necrosis in flakes accompanied by evident hemorrhage were documented. Conclusions The D-Gal dosage in the medium-and high-dose groups meet the standards of the ALF model. The degree of ALF in the medium-dose group is relatively slight, which is beneficial to the implementation of liver transplantation. ALF in the high-dose group is relatively severe, which is suitable for the evaluation of the clinical efficacy of therapeutic options.
Objective To evaluate the effect of different doses of D-galactosamine(D-Gal) on non-human primate cynomolgus monkey and to establish a monkey model with different degree of acute liver failure(ALF). Methods Twelve cynomolgus monkeys were evenly divided into the low-, medium-and high-dose groups(n=4) with a dosage of 0.23, 0.25 and 0.27 g/kg, respectively. In each group, the corresponding dose of D-Gal solution was injected into the monkeys through the forearm vein at one time in a sober state(without anesthesia). The survival time of the cynomolgus monkeys was recorded. Digestive tract and hepatic encephalopathy symptoms were observed. Vital signs were measured at 0 h before and 12, 24, 36, 48, 60, 72, 96, 120 and 144 h after D-Gal administration. Alanine transaminase(ALT), total bilirubin(TB), prothrombin time(PT), blood ammonia and other parameters were detected from the blood samples. The liver tissues were prepared for hematoxylin-eosin(HE) staining to observe the pathological changes. Results All cynomolgus monkeys in the low-dose group survived and transient liver injury was noted without the hepatic encephalopathy symptoms. At 60 h after D-Gal administration, the liver function and coagulation indexes reached the peak, gradually recovered and then basically returned to the normal range at 120 h. In the medium-dose group, the course of disease was relatively slow and gradually recovered after the appearance of severe liver damage and hepatic encephalopathy symptoms and only one animal died. All cynomolgus monkeys in the high-dose group died after developing hepatic encephalopathy symptoms and severe liver damage with a mean survival time of(72±13) h. Pathological examination of liver tissue demonstrated that scattered liver cell necrosis and inflammatory cell infiltration were observed in the liver tissues of the low-dose group. In the medium-and high-dose groups, the hepatic lobule structure was not clear, and the liver cell necrosis in flakes accompanied by evident hemorrhage were documented. Conclusions The D-Gal dosage in the medium-and high-dose groups meet the standards of the ALF model. The degree of ALF in the medium-dose group is relatively slight, which is beneficial to the implementation of liver transplantation. ALF in the high-dose group is relatively severe, which is suitable for the evaluation of the clinical efficacy of therapeutic options.
引文
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[17]BUTTERWORTH RF.Pathogenesis of hepatic encephalopathy and brain edema in acute liver failure[J].J Clin Exp Hepatol,2015,5(Suppl 1):S96-S103.DOI:10.1016/j.jceh.2014.02.004.
[18]曾艳丽,高飞,魏君峰,等.血栓弹力图评价慢加急性肝衰竭患者的凝血功能及预后研究[J].中华肝脏病杂志,2017,25(1):32-37.DOI:10.3760/cma.j.issn.1007-3418.2017.01.008.ZENG YL,GAO F,WEI JF,et al.Value of thromboelastography in evaluating coagulation function and prognosis in patients with acute-on-chronic liver failure[J].Chin JHepatol,2017,25(1):32-37.DOI:10.3760/cma.j.issn.1007-3418.2017.01.008.
[19]RAMANATHAN R,PETTINATO G,BEESTON JT,et al.Transplantation of human stem cell-derived hepatocytes in an animal model of acute liver failure[J].Surgery,2015,158(2):349-359.DOI:10.1016/j.surg.2015.04.014.
[2]CARDOSO FS,MARCELINO P,BAGULHO L,et al.Acute liver failure:an up-to-date approach[J].J Crit Care,2017,39:25-30.DOI:10.1016/j.jcrc.2017.01.003.
[3]ISIK A,FIRAT D.Bilateral intra-areolar polythelia[J].Breast J,2018,24(1):89-90.DOI:10.1111/tbj.12838.
[4]DEFTEREVOS G,NASTOS C,PAPALOIS A,et al.Peritoneal albumin dialysis as a novel approach for liver support:study in a porcine model of acute hepatic failure[J].Artif Organs,2016,40(8):755-764.DOI:10.1111/aor.12687.
[5]SANG JF,SHI XL,HAN B,et al.Intraportal mesenchymal stem cell transplantation prevents acute liver failure through promoting cell proliferation and inhibiting apoptosis[J].Hepatobiliary Pancreat Dis Int,2016,15(6):602-611.
[6]桑剑锋,马虎成,施晓雷,等.巴马小型猪85%肝切除术后急性肝功能衰竭模型的建立[J].中国实验动物学报,2017,25(2):174-180.DOI:10.3969/j.issn.1005-4847.2017.02.011.SANG JF,MA HC,SHI XL,et al.Establishment of a porcine model of acute liver failure after 85%hepatectomy[J].Acta Lab Anim Sci Sin,2017,25(2):174-180.DOI:10.3969/j.issn.1005-4847.2017.02.011.
[7]FLAMM SL,YANG YX,SINGH S,et al.American gastroenterological association institute guidelines for the diagnosis and management of acute liver failure[J].Gastroenterology,2017,152(3):644-647.DOI:10.1053/j.gastro.2016.12.026.
[8]FENG L,CAI L,HE GL,et al.Novel D-galactosamineinduced cynomolgus monkey model of acute liver failure[J].World J Gastroenterol,2017,23(42):7572-7583.DOI:10.3748/wjg.v23.i42.7572.
[9]CAI L,WENG J,FENG L,et al.Establishment of a novel simplified surgical model of acute liver failure in the cynomolgus monkey[J].Biomed Res Int,2016:3518989.DOI:10.1155/2016/3518989.
[10]时红波,时红林,张向颖,等.细胞自噬在D-氨基半乳糖/脂多糖诱导的小鼠急性肝损伤模型中的保护作用及机制[J].临床肝胆病杂志,2017,33(2):329-333.DOI:10.3969/j.issn.1001-5256.2017.02.027.SHI HB,SHI HL,ZHANG XY,et al.Protective effect of autophagy in mice with acute liver injury induced by D-galactosamine/lipopolysaccharide and related mechanisms[J].J Clin Hepatol,2017,33(2):329-333.DOI:10.3969/j.issn.1001-5256.2017.02.027.
[11]HE GL,FENG L,CAI L,et al.Artificial liver support in pigs with acetaminophen-induced acute liver failure[J].World J Gastroenterol,2017,23(18):3262-3268.DOI:10.3748/wjg.v23.i18.3262.
[12]NING Y,KIM JK,MIN HK,et al.Cholesterol metabolites alleviate injured liver function and decrease mortality in an LPS-induced mouse model[J].Metabolism,2017,71:83-93.DOI:10.1016/j.metabol.2016.12.007.
[13]TU?óN MJ,ALVAREZ M,CULEBRAS JM,et al.An overview of animal models for investigating the pathogenesis and therapeutic strategies in acute hepatic failure[J].World JGastroenterol,2009,15(25):3086-3098.
[14]MAES M,VINKEN M,JAESCHKE H.Experimental models of hepatotoxicity related to acute liver failure[J].Toxicol Appl Pharmacol,2016,290:86-97.DOI:10.1016/j.taap.2015.11.016.(下转第页)
[15]GLORIOSO JM,MAO SA,RODYSILL B,et al.Pivotal preclinical trial of the spheroid reservoir bioartificial liver[J].J Hepatol,2015,63(2):388-398.DOI:10.1016/j.jhep.2015.03.021.
[16]ZHOU P,SHAO L,ZHAO L,et al.Efficacy of fluidized bed bioartificial liver in treating fulminant hepatic failure in pigs:a metabolomics study[J].Sci Rep,2016,6:26070.DOI:10.1038/srep26070.
[17]BUTTERWORTH RF.Pathogenesis of hepatic encephalopathy and brain edema in acute liver failure[J].J Clin Exp Hepatol,2015,5(Suppl 1):S96-S103.DOI:10.1016/j.jceh.2014.02.004.
[18]曾艳丽,高飞,魏君峰,等.血栓弹力图评价慢加急性肝衰竭患者的凝血功能及预后研究[J].中华肝脏病杂志,2017,25(1):32-37.DOI:10.3760/cma.j.issn.1007-3418.2017.01.008.ZENG YL,GAO F,WEI JF,et al.Value of thromboelastography in evaluating coagulation function and prognosis in patients with acute-on-chronic liver failure[J].Chin JHepatol,2017,25(1):32-37.DOI:10.3760/cma.j.issn.1007-3418.2017.01.008.
[19]RAMANATHAN R,PETTINATO G,BEESTON JT,et al.Transplantation of human stem cell-derived hepatocytes in an animal model of acute liver failure[J].Surgery,2015,158(2):349-359.DOI:10.1016/j.surg.2015.04.014.
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