HO-1在大鼠胆道缺血再灌注损伤中对胆盐转运蛋白的影响及机制研究
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摘要
目的：
本研究通过调节肝脏血红素氧合酶-1(HO-1)的表达,来观察其对肝脏胆盐转运蛋白表达的影响,探讨其对大鼠胆道上皮细胞缺血再灌注损伤(IRI)的作用及其可能机制。
方法：
1.采用Fei X等报道的方法操作,建立70%肝脏缺血再灌注损伤模型。
2.使用化学合成的Ad-HO-1、Ad-HO-1-siRNA,以及空腺病毒载体和生理盐水,经阴茎背静脉注射导入大鼠肝脏,24小时后开腹,建立大鼠肝脏缺血再灌注损伤模型。动物随机分为4组：生理盐水对照组(n=32)；空载体组(n=32)；HO-1组(n=32),注射剂量为2×109TU/只,MOI=1；siRNA组(n=32),注射剂量为2×109TU/只,MOI=1。各组再分别于再灌注损伤后1h、1d、1w和2w处死8只大鼠。检测肝脏缺血再灌注后血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TB)、碱性磷酸酶(ALP)水平；ELISA法检测IL-6IL-10、HGF、TNF-a在血清中的水平；荧光定量PCR检测胆盐输出泵(Bsep) mRNA、多药耐药蛋白2(Mrp2) mRNA和牛磺胆酸钠共转运蛋白(Ntcp) mRNA的表达,以及肝组织1d时相点HO-1的mRNA表达；Western Blotting检测肝组织Ntcp蛋白的表达；肝脏及肝门部胆管组织标本行免疫荧光双染及氯醋酸酯酶染色,观察大小胆管及周围肝组织形态结构的变化。
结果：
1.共成功实行大鼠肝脏缺血再灌注损伤模型128例,肝脏部分缺血时间为45min。
2.相同时相点比较,HO-1组血清ALT、AST、TB、ALP低于对照组和空载体组(p<0.05)：siRNA组血清ALT、AST、TB、ALP含量高于对照组和空载体组(p<0.05)；HO-1组明显低于siRNA组(p<0.01),有统计学意义。
3.相同时相点比较, siRNA组血清HGF、IL-10明显低于对照组和空载体组(p<0.05)；HO-1组血清HGF、IL-10明显高于对照组和空载体组(p<0.05)；HO-1组明显高于siRNA组(p<0.01),有统计学意义。HO-1组血清AIL-6、TNF-a低于对照组和空载体组(p<0.05)；siRNA组血清IL-6、TNF-a含量高于对照组和空载体组(p<0.05)；HO-1组明显低于siRNA组(p<0.01),有统计学意义。
4.相同时相点比较,HO-1组HO-1、Ntcp、Bsep、Mrp2mRNA表达水平高于对照组和空载体组(p<0.05)；siRNA组HO-1、Ntcp、Bsep、Mrp2mRNA表达水平低于对照组和空载体组(p<0.05)；HO-1组表达水平明显高于siRNA组(p<0.01),有统计学意义。
5.相同时相点比较,siRNA组Ntcp蛋白表达水平低于对照组和空载体组(p<0.05);HO-1组Ntcp蛋白表达水平高于对照组和空载体组(p<0.05)；HO-1组表达水平明显高于siRNA组(p<0.01),有统计学意义。
6.相同时相点比较,siRNA组免疫荧光双染及氯醋酸酯酶染色上可见：大小胆管上皮细胞模糊,部分胆管基底膜脱落,周围肝细胞松散,伴核变性,严重损伤表现；HO-1组损伤表现较轻,大小胆管上皮细胞清晰,基底膜的连续性可,胆管周围肝细胞排列紧密,未见核变性。
结论：
1.大鼠肝脏热缺血再灌注损伤模型是一种稳定、可靠、简便的模型,可操作性强。
2. Ad-HO-1能有效增加HO-1基因在大鼠肝脏的表达,而Ad-HO-1-siRNA能有效减少HO-1基因在大鼠肝脏的表达。
3.缺血再灌注会造成肝功能受损,Ad-HO-1可有效改善肝功能,减少促炎性细胞因子IL-6、TNF-a的表达,增加了保护性细胞因子HGF、IL-10的表达,减轻了大、小胆管的缺血再灌注损伤。
4.HO-1保护大鼠胆道缺血再灌注损伤的机制,可能与诱导胆盐转运蛋白高表达有关；胆盐转运蛋白高表达可以促进胆汁的转运和排泄,从而减少具有较强细胞毒性的“毒性胆汁”形成,最终减少胆盐对胆管的损伤。
Objective
In this study, by regulating the expression of liver heme oxygenase-1(HO-1), to observe the expression of hepatic bile salt transport protein by HO-1, Explore the rat biliary epithelial cell in ischemia and reperfusion injury (IRI) and its possible mechanisms
Methods
1. To establish the models of70%part of the liver ischemia-reperfusion injury by the method of operation which Fei X had reported.
2. The use of chemical synthesis of Ad-HO-1, Ad-HO-1-of siRNA, empty adenoviruses and saline, into the donor liver by Genital dorsal vein. laparotomy after24hours, establish rat liver ischemia-reperfusion injury model. The animals were randomly divided into four groups:NaCl group, Empty adenovirus group, Amplification of HO-1adenovirus group, Silencing HO-1adenovirus group. To Collect specimens on1hour,1day,1week and2weeks after operation. To detect the levels of ALT、AST、TB、ALP in serum after ischemia reperfusion injury of liver. The function of liver was also evaluated. The expressions of IL-6、IL-10、HGF and TNF-a in the liver were detected. The mRNA and protein expressions of salt transporter proteins(Sodium taurocholate cotransporting polypeptide, Bile salt expot pump、Multidrug resistance-associated protein-2、sodium taurocholate cotransporting polypeptide and the HO-1mRNA expressions of liver tissue of levels in1d were measured. The livere and biliary bile duct were detected by immunofluorescence double staining and Chloro-acetate esterase staining. Observe the morphological changes in the structure of the size of the bile duct and the surrounding liver tissue.
Results
1.128cases of ischemia-reperfusion injury models of liver were Successfully established and partial liver ischemia time was45minutes.
2. Compared with the same time points, serum ALT, AST, TB, and the ALP in HO-1groups was significantly lower than in NaCl and Adenovirus groups (p<0.05). serum ALT, AST, TB, and the ALP in siRNA groups was significantly higher than in NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly lower than the siRNA groups (p<0.01).
3. Compared with the same time points, siRNA groups serum HGF and IL-10was significantly lower than in NaCl and Adenovirus groups (p<0.05); HO-1groups serum HGF and IL-10was significantly higher than in NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly higher than the siRNA groups (p<0.01). serum IL-6and TNF-a in G3was significantly lower than in NaCl and Adenovirus groups (p<0.05). serum IL-6and TNF-a in siRNA groups was significantly higher than in NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly lower than the siRNA groups (p<0.01).
4. Compared with the same time points, HO-1groups of HO-1, Ntcp, Bsep, Mrp2mRNA expression level was significantly higher than in NaCl and Adenovirus groups (p<0.05); siRNA groups of HO-1, Ntcp, Bsep, Mrp2mRNA expression level was significantly lower than in NaCl and Adenovirus groups (p<0.05); HO-1groups was significantly higher than the siRNA groups (p<0.01).
5. siRNA groups Ntcp protein expression levels were significantly lower than the NaCl and Adenovirus groups (p<0.05). HO-1groups Ntcp protein expression levels were significantly higher than the NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly higher than the siRNA groups (p<0.01).
6. Compared with the same time points, Immunofluorescence double staining and chloro-acetate esterase staining in siRNA groups can be seen:The size of the bile duct epithelial cells is fuzzy, part of the bile duct basement membrane shedding, liver cells around loose, with degeneration, serious injury performance. The HO-1groups was less severe injury, size of the bile duct epithelial cells is clear, the continuity of the basement membrane is better, around the bile duct liver cells closely arranged, no degeneration
Conclusions:
1. Rat liver warm ischemia reperfusion injury model is a stable, reliable, simple, the model is workable.
2. Ad-HO-1can effectively increase HO-1gene expression in rat liver; Ad-HO-1-siRNA can effectively reduce the expression of HO-1gene in rat liver.
3. The liver function damaged by Ischemia and reperfusion. Ad-HO-1can effectively improve liver function. Reduce the proinflammatory cytokine IL-6, TNF-a expression, an increase of protective cytokines of HGF, IL-10expression. Reduce the size of the bile duct ischemia and reperfusion injury.
4. The mechanism of HO-1protect biliary ischemia-reperfusion injury in rats, may be induced by bile salt transporter protein high expression. Bile salt transporter protein expression can contribute to the transport and excretion of bile, reduce the strong cytotoxic bile formation, ultimately reduce the role of bile salts of the bile duct.
本研究通过调节肝脏血红素氧合酶-1(HO-1)的表达,来观察其对肝脏胆盐转运蛋白表达的影响,探讨其对大鼠胆道上皮细胞缺血再灌注损伤(IRI)的作用及其可能机制。
方法：
1.采用Fei X等报道的方法操作,建立70%肝脏缺血再灌注损伤模型。
2.使用化学合成的Ad-HO-1、Ad-HO-1-siRNA,以及空腺病毒载体和生理盐水,经阴茎背静脉注射导入大鼠肝脏,24小时后开腹,建立大鼠肝脏缺血再灌注损伤模型。动物随机分为4组：生理盐水对照组(n=32)；空载体组(n=32)；HO-1组(n=32),注射剂量为2×109TU/只,MOI=1；siRNA组(n=32),注射剂量为2×109TU/只,MOI=1。各组再分别于再灌注损伤后1h、1d、1w和2w处死8只大鼠。检测肝脏缺血再灌注后血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TB)、碱性磷酸酶(ALP)水平；ELISA法检测IL-6IL-10、HGF、TNF-a在血清中的水平；荧光定量PCR检测胆盐输出泵(Bsep) mRNA、多药耐药蛋白2(Mrp2) mRNA和牛磺胆酸钠共转运蛋白(Ntcp) mRNA的表达,以及肝组织1d时相点HO-1的mRNA表达；Western Blotting检测肝组织Ntcp蛋白的表达；肝脏及肝门部胆管组织标本行免疫荧光双染及氯醋酸酯酶染色,观察大小胆管及周围肝组织形态结构的变化。
结果：
1.共成功实行大鼠肝脏缺血再灌注损伤模型128例,肝脏部分缺血时间为45min。
2.相同时相点比较,HO-1组血清ALT、AST、TB、ALP低于对照组和空载体组(p<0.05)：siRNA组血清ALT、AST、TB、ALP含量高于对照组和空载体组(p<0.05)；HO-1组明显低于siRNA组(p<0.01),有统计学意义。
3.相同时相点比较, siRNA组血清HGF、IL-10明显低于对照组和空载体组(p<0.05)；HO-1组血清HGF、IL-10明显高于对照组和空载体组(p<0.05)；HO-1组明显高于siRNA组(p<0.01),有统计学意义。HO-1组血清AIL-6、TNF-a低于对照组和空载体组(p<0.05)；siRNA组血清IL-6、TNF-a含量高于对照组和空载体组(p<0.05)；HO-1组明显低于siRNA组(p<0.01),有统计学意义。
4.相同时相点比较,HO-1组HO-1、Ntcp、Bsep、Mrp2mRNA表达水平高于对照组和空载体组(p<0.05)；siRNA组HO-1、Ntcp、Bsep、Mrp2mRNA表达水平低于对照组和空载体组(p<0.05)；HO-1组表达水平明显高于siRNA组(p<0.01),有统计学意义。
5.相同时相点比较,siRNA组Ntcp蛋白表达水平低于对照组和空载体组(p<0.05);HO-1组Ntcp蛋白表达水平高于对照组和空载体组(p<0.05)；HO-1组表达水平明显高于siRNA组(p<0.01),有统计学意义。
6.相同时相点比较,siRNA组免疫荧光双染及氯醋酸酯酶染色上可见：大小胆管上皮细胞模糊,部分胆管基底膜脱落,周围肝细胞松散,伴核变性,严重损伤表现；HO-1组损伤表现较轻,大小胆管上皮细胞清晰,基底膜的连续性可,胆管周围肝细胞排列紧密,未见核变性。
结论：
1.大鼠肝脏热缺血再灌注损伤模型是一种稳定、可靠、简便的模型,可操作性强。
2. Ad-HO-1能有效增加HO-1基因在大鼠肝脏的表达,而Ad-HO-1-siRNA能有效减少HO-1基因在大鼠肝脏的表达。
3.缺血再灌注会造成肝功能受损,Ad-HO-1可有效改善肝功能,减少促炎性细胞因子IL-6、TNF-a的表达,增加了保护性细胞因子HGF、IL-10的表达,减轻了大、小胆管的缺血再灌注损伤。
4.HO-1保护大鼠胆道缺血再灌注损伤的机制,可能与诱导胆盐转运蛋白高表达有关；胆盐转运蛋白高表达可以促进胆汁的转运和排泄,从而减少具有较强细胞毒性的“毒性胆汁”形成,最终减少胆盐对胆管的损伤。
Objective
In this study, by regulating the expression of liver heme oxygenase-1(HO-1), to observe the expression of hepatic bile salt transport protein by HO-1, Explore the rat biliary epithelial cell in ischemia and reperfusion injury (IRI) and its possible mechanisms
Methods
1. To establish the models of70%part of the liver ischemia-reperfusion injury by the method of operation which Fei X had reported.
2. The use of chemical synthesis of Ad-HO-1, Ad-HO-1-of siRNA, empty adenoviruses and saline, into the donor liver by Genital dorsal vein. laparotomy after24hours, establish rat liver ischemia-reperfusion injury model. The animals were randomly divided into four groups:NaCl group, Empty adenovirus group, Amplification of HO-1adenovirus group, Silencing HO-1adenovirus group. To Collect specimens on1hour,1day,1week and2weeks after operation. To detect the levels of ALT、AST、TB、ALP in serum after ischemia reperfusion injury of liver. The function of liver was also evaluated. The expressions of IL-6、IL-10、HGF and TNF-a in the liver were detected. The mRNA and protein expressions of salt transporter proteins(Sodium taurocholate cotransporting polypeptide, Bile salt expot pump、Multidrug resistance-associated protein-2、sodium taurocholate cotransporting polypeptide and the HO-1mRNA expressions of liver tissue of levels in1d were measured. The livere and biliary bile duct were detected by immunofluorescence double staining and Chloro-acetate esterase staining. Observe the morphological changes in the structure of the size of the bile duct and the surrounding liver tissue.
Results
1.128cases of ischemia-reperfusion injury models of liver were Successfully established and partial liver ischemia time was45minutes.
2. Compared with the same time points, serum ALT, AST, TB, and the ALP in HO-1groups was significantly lower than in NaCl and Adenovirus groups (p<0.05). serum ALT, AST, TB, and the ALP in siRNA groups was significantly higher than in NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly lower than the siRNA groups (p<0.01).
3. Compared with the same time points, siRNA groups serum HGF and IL-10was significantly lower than in NaCl and Adenovirus groups (p<0.05); HO-1groups serum HGF and IL-10was significantly higher than in NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly higher than the siRNA groups (p<0.01). serum IL-6and TNF-a in G3was significantly lower than in NaCl and Adenovirus groups (p<0.05). serum IL-6and TNF-a in siRNA groups was significantly higher than in NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly lower than the siRNA groups (p<0.01).
4. Compared with the same time points, HO-1groups of HO-1, Ntcp, Bsep, Mrp2mRNA expression level was significantly higher than in NaCl and Adenovirus groups (p<0.05); siRNA groups of HO-1, Ntcp, Bsep, Mrp2mRNA expression level was significantly lower than in NaCl and Adenovirus groups (p<0.05); HO-1groups was significantly higher than the siRNA groups (p<0.01).
5. siRNA groups Ntcp protein expression levels were significantly lower than the NaCl and Adenovirus groups (p<0.05). HO-1groups Ntcp protein expression levels were significantly higher than the NaCl and Adenovirus groups (p<0.05). HO-1groups was significantly higher than the siRNA groups (p<0.01).
6. Compared with the same time points, Immunofluorescence double staining and chloro-acetate esterase staining in siRNA groups can be seen:The size of the bile duct epithelial cells is fuzzy, part of the bile duct basement membrane shedding, liver cells around loose, with degeneration, serious injury performance. The HO-1groups was less severe injury, size of the bile duct epithelial cells is clear, the continuity of the basement membrane is better, around the bile duct liver cells closely arranged, no degeneration
Conclusions:
1. Rat liver warm ischemia reperfusion injury model is a stable, reliable, simple, the model is workable.
2. Ad-HO-1can effectively increase HO-1gene expression in rat liver; Ad-HO-1-siRNA can effectively reduce the expression of HO-1gene in rat liver.
3. The liver function damaged by Ischemia and reperfusion. Ad-HO-1can effectively improve liver function. Reduce the proinflammatory cytokine IL-6, TNF-a expression, an increase of protective cytokines of HGF, IL-10expression. Reduce the size of the bile duct ischemia and reperfusion injury.
4. The mechanism of HO-1protect biliary ischemia-reperfusion injury in rats, may be induced by bile salt transporter protein high expression. Bile salt transporter protein expression can contribute to the transport and excretion of bile, reduce the strong cytotoxic bile formation, ultimately reduce the role of bile salts of the bile duct.
引文
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[33]李国钦.肝细胞刺激因子能否刺激肝再生的研究[J].中西医结合肝病杂志,2000,10(2)：59-61
[34]林梅.肝源性促肝细胞生长因子的研究与临床现状[J].中国实用内科学杂志,1997,17(1)：56-57.
[35]Schmidt C, Bladt F, et,al. Scatter factor/hepatocyte growth factor isessential for liver development[J]. Nature,1995,373(6516):699-702.
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[37]Kamimoto M, Mizuno S, Matsumoto K, et,al. Hepatocyte growth factor prevents multiple organ injuries in endotoxemic mice through a heme oxygenase-1-dependent mechanism. Biochem Biophys Res Commun.2009 Mar 6;380(2):333-7.
[38]Kamimoto M;Mizuno S;Nakamura T,et,al. Reciprocal regulation of IL-6 and IL-10 balance by HGF via recruitment of heme oxygenase-1 in macrophages for attenuation of liver injury in a mouse model of endotoxemia. Int J Mol Med.2009V24N2:161-70
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[2]Tenhunen R, Marver HS, Schmid R. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase[J]. Proc Natl Acad Sci USA,1968,61: 748-755.
[3]MD Maines, GM Trakshel, and RK Kutty Characterization of two constitutive froms of rat liver microsomal heme oxygenase. Only one molecular species of the enzyme is inducibleJ.Biol.Chem.1986 261:411-419.
[4]Trakshel GM, Maines MD. Multiplicity of heme oxygenase isozymes.HO-1 and HO-2 are different molecular species in rat and rabbit. J. biol.Chem.1989 264:1323-1328.
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[6]Wise CD, Drabkin DL. Degradation of haemoglobin and hemin to biliverdin by a new cell-free system obtained from the hemophagous organ of dog placenta. Fed Proc 23:323.
[7]Llesuy SF, Tomaro ML Heme oxygenase and oxidative stress. Evidence of involvement of bilirubin as Physiological protector against oxidative damage. Biochim. BioPhys. Acta 1994,1223(1):9-14.
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[24]Zhong Zeng, Han-Fei Huang,at al. Heme oxygenase-1 protects donor livers from ischemia/reperfusion injury:The role of kupffer cells. World J Gastroenterol 2010 March 14;16(10):1285-1292.
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[1]Amesri F, Buelow R, Kato H, et al. Upregulation of heme oxygenase-1 protects genetically fat Zucker rat livers from ischemia/reperfusion injury[J]. J Clin Invest,1999,104:1631-1639.
[2]Tenhunen R, Marver HS, Schmid R. The enzymatic conversion of heme to bilirubin by microsomal heme oxygenase[J]. Proc Natl Acad Sci USA,1968,61: 748-755.
[3]MD Maines, GM Trakshel, and RK Kutty Characterization of two constitutive froms of rat liver microsomal heme oxygenase. Only one molecular species of the enzyme is inducibleJ.Biol.Chem.1986 261:411-419.
[4]Trakshel GM, Maines MD. Multiplicity of heme oxygenase isozymes.HO-1 and HO-2 are different molecular species in rat and rabbit. J. biol.Chem.1989 264:1323-1328.
[5]McCoubrey W, Huang T, Maines M. Isolation and characterization of a cDNA from the rat brain thatencodes hemoProtein heme oxygenase-3.Eur J Bioehem 1997;247:725-732.
[6]Wise CD, Drabkin DL. Degradation of haemoglobin and hemin to biliverdin by a new cell-free system obtained from the hemophagous organ of dog placenta. Fed Proc 23:323.
[7]Llesuy SF, Tomaro ML Heme oxygenase and oxidative stress. Evidence of involvement of bilirubin as Physiological protector against oxidative damage. Biochim. BioPhys. Acta 1994,1223(1):9-14.
[8]Maines MD. The heme oxygenase system:a regulator of second messenger gases[J].Annu Rve Phamraeol Toxciol,1997,37:517-554.
[9]Abrahm NG, Lavrovsky Y, Schwartzman ML, et al. Transfection of the human heme oxygenase gene into rabbit coronary microvessel endothelial cells: protective effect against heme and hemoglobin toxicity[J].Proc Natl Acad Sci USA, 1995,92:6798-6802.
[10]Teyrr CM, Clikeman JA, Hoidal JR, and Callahan KS. TNF-alpha and IL-lalpha induce heme oxygenas-1 via protein kinase C, Ca2+and phospholiPase A2 in endothelial cells[J]. Am J Physiol,1999,276:H1493-H501.
[11]Immenschuh S, Kietzmann T, Hinke V, et al. te rat heme oxygenase-1 gene is transcriptionally induced via the protein kinase A signaling pathway in rat hepatocyte cultures [J]. Mol Pharmacol,1998,53:483-491.
[12]Polte T, Abate A, Dennery PA, and Schroder H.Heme oxygenase-1 is a eGMP-inducible endothelial Protein and mediates the cytoprotective action of nitric oxide[J]. Arterioscler Thromb Vasc Biol,2000,20:1209-1215.
[13]Johnson RK, Kozma F, and Colomlbari E. Carbon monoxide:from toxin to endogenous modulator of cardiovascular functions[J]. Braz J Med Biol Res,1999,32: 1-14.
[14]田飞,张黎,郭琼行,等.血红素氧合酶-1及TOLL样受体2/4在直肠癌组织 中的表达及意义[J].中国现代普通外科进展,2008,11(3)：253-254.
[15]Stoker R, Yamamoto Y, Mcdonagh AF, et al. Bilirubin is an antioxidant of possible Physiologic importance[J]. Seienec,1987,235(4792):1043-1046.
[16]Naziroglu M, Cay M, Ustundag B, et al. Portective effects of Vitmin E on carbon Tetrachloride-induced liver damage in ratIJ]. Cell Biochem Funct,1999,17(4): 253-259.
[17]Siwo RC, Sato H, Leake DS, et al. Vitamin C protects human arterial smooth cells against atherogenic lipoproteins:effects of antioxidant Vitamins on oxidized LDL-induced increase in cystin transport and glutathione. [J]. Arterioscler Thromb Vasc Biol,1998,18(10):1662-1670.
[18]Mayer M. Association of serum bilirubin concent ration with risk of coronary artery disease[J]. Clin Chem,2000,46(11):1723.
[19]Soares M P,Brouard S, Smith RN, er al. Heme oxygenase-l,a protective gene that prevents the rejection of transplanted organs. Immunol Rev,2001, 184:275-285.
[20]Durante W. Carbon monoxide and bile pigments:surprising mediators of vascular function. Vasc Med,2002,7(3):195-202.
[21]Berberat P. O, Katori M, Kaczmarek E, et, al.Heavy chain ferritin acts as an antiapoptotic gene that protects livers from ischemia reperfusion injury.FASEB J.2003,17:1724.
[22]Ferris CD, Jaffrey SR, Sawa A, at al. Heme oxygenase-1 prevents cell death by regulating cellular iron. Nat. Cell Biol 1999,1(3):152-157.
[23]Wagener F A, Eggert A, Boerman O C, at al. Heme is a potent inducer of inflammation in mice and is counteracted by heme oxygenase[J]. Blood,2001,98(6): 1802-1811.
[24]Zhong Zeng, Han-Fei Huang,at al. Heme oxygenase-1 protects donor livers from ischemia/reperfusion injury:The role of kupffer cells. World J Gastroenterol 2010 March 14;16(10):1285-1292.
[25]Devey L, Ferenbach D, Mohr E,et al. Tissue-resident macrophages protect the liver from ischemia reperfusion injury via a heme oxygenase-1 dependent mechanism. Mol Ther,2009,17(1);65-72.
[26]Kobayashi T, Hirano K, Yamamoto T, et al.The protective role of kupffer cell in the ischemia-reperfusion rat liver. Archives of Histology & Cytology,2002, 65(3):251-261.
[27]De Broe ME, Bonventre JV,Molitoris BA, et al. Evolving basic concepts in ischemia injury. Kidney Int.2004,66:479-531.
[28]王万铁,林丽娜,谢克俭,等异丙酚对兔肝缺血/再灌注损伤中一氧化碳和内皮素的干预.中国急救医学,2004,24(1)：4-6.
[29]Z. Zeng, H.F. Huang,at al. Contributions of Heme Oxygenase-1 in Postconditioning-Protected Ischemia-Reperfusion Injury in Rat Liver Transplantation. Transplant Proc.2011V43N7:2517-23
[30]Amersi F, Shen XD,Anselmo D, et al.Ex vivo exposure to carbon monoxide prevents hepatic ischemia/reperfusion injury through p38MAP kinase pathway[J]. Hepatology,2002,35(4):815-823.
[31]Brouard S, Berberat PO, Tobiasch E,et al.Heme oxygenase-1 derived carbon monoxide requires the activation of the transcription factor NF-kB to protect endothelial cells from TNF-alpha mediated apoptosis[J]. Biol Chem,2002,277: 17950-17961.
[32]Duckers,Davidson CA, Kaminski PM,Fayngersh RP, and Mohazzab HK. Oxidant-nitric oxide signalling mechanisms in vascular tissue[J]. Biochemistry(Mosc),1998,63:810-816.
[33]Togane Y, Morita T, Suematsu M, at al. Protective roles of endogenous carbon monoxide in neointimal development elicited by arterial injury. Am J Physiol Heart Cic Physiol 2000,278(2):H623-632.
[34]Moore KW, de Waal Malefyt R, Coffman RL,et al. Interleukin-10 and the interleukin-10 receptor. Annu. Rev. Immunol.2001,19:683.
[35]Lee TS,Chau LY. Heme oxygenase-1 mediates the antiinflammatory effect of interleukin-10 in mice. Nature Med.2002,8:240.
[36]Soares MP.VEGF:is it just an inducer of heme oxygenase-1 expression. Blood,2004,103:751.
[37]Z Zeng, H F Huang, M Q Chen, et al. Postconditioning Prevents I/R Injury in Liver Transplantation. Hepato-Gastroenterology 2010,57:1-6.