目的:研究活血化瘀注射液I号(HHI-I)预处理与缺血预处理(ischemic preconditioning,IP)对大鼠肝脏缺血再灌注(ischemia and reperfusion, I/R)损伤的改善作用,并比较两者的作用效果．方法:健康♂SD大鼠80只,随机分为假手术对照组(Shaman...目的:研究活血化瘀注射液I号(HHI-I)预处理与缺血预处理(ischemic preconditioning,IP)对大鼠肝脏缺血再灌注(ischemia and reperfusion, I/R)损伤的改善作用,并比较两者的作用效果．方法:健康♂SD大鼠80只,随机分为假手术对照组(Shaman)、缺血再灌注组(I／R组)、缺血预处理组(IP组)、HHI-I预处理组(HHI-I组),每组20只．建立大鼠部分肝缺血模型,各组在I／R后1,3,6,24 h分别取材,测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)的水平:取左肝测组织丙二醛(MDA)、超氧化物歧化酶(SOD)的含量．I／R后1 h RT- PCR检测肝组织中肿瘤坏死因子α(TNF-α)和细胞间黏附分子-1(ICAM-1)mRNA的表达, I／R后3 h行组织学观察．结果:I／R组、IP组、HHI-I组的ALT,AST, LDH活性、MDA值和TNF-α．ICAM-1 mRNA表达水平均明显高于Sham组．IP组、HHI-I组低于I／R组．HHI-I组所有时间点ALT,AST, LDH明显低于IP组(ALT:2378．8±303．4 nkat／L VS 2840．6±248．4 nkat／L;AST:2887．2±270．1 nkat／L vs 4567．6±275．1 nkat／L;LDH:10550．4±710．1 nkat／L vs 12164．1±735．1 nkat／L;P均<0．05)．HHI-I组MDA值明显低于IP组(17．35±1．39 nmol／g vs 21．66±1．84 nmol／g,P<0．05)．HHI-I组TNF-α,ICAM-1 mRNA表达水平低于IP组(TNF-α:0．54±0．06 vs 0．78±0．08; ICAM-1:0．43±0．03 vs 0．69±0．11,P均<0．01)．各组SOD值均低于Sham组(P<0．05)．IP组、HHI-I组均高于I／R组(P<0．05),HHI-I组SOD(1．3,6 h)明显高于IP组(136．00±12．50 nmol/gvs 124．70±9．32 nmol／g,P<0．05)．Sham组光镜下肝小叶结构正常;I/R组肝小叶结构紊乱,肝细胞水肿变性;IP组肝细胞水肿明显,部分肝细胞变性;HHI-I组肝小叶结构基本正常,肝细胞无明显水肿．结论:HHI-I预处理与IP均可改善I／R对肝脏造成的损伤,前者的效果优于后者．HHI-I的保护机制可能在于改善肝脏微循环,减轻组织缺氧状态,并通过抑制TNF-α和ICAM-1等细胞因子和细胞黏附�展开更多
Objective To investigate the role of intercellular adhesion molecule-1 (ICAM-1) andP- selectin in hepatic ischemia/reperfusion injury. Methods The relationship between P- selectin as well asICAM- 1 and liver ischemia/...Objective To investigate the role of intercellular adhesion molecule-1 (ICAM-1) andP- selectin in hepatic ischemia/reperfusion injury. Methods The relationship between P- selectin as well asICAM- 1 and liver ischemia/reperfusion was studied, using a 60 minutes ischemia - reperfusion rat liver model.The animals were divided into three groups including: the sham group, the ischemic control group receiving onlythe normal saline and the treated group receiving anti-P-selection monoclonal antibody (Mab) at a dose of2mg/kg 15 minutes before reperfusion. The following indexes were analyzed: liver injury tests, liver histotogy,serum enzymes level (AST, ALT), and the levels of ICAM-1 and P-selectin in the blood of rats. ResultsSerum enzymes levels were significantly increased during sixty minutes of left lobar ischemia and reperfusion(saline control), and the increment was significantly inhibited with P- selectin Mab. Liver pathology observed bylight microscopy was greatly ameliorated by Mab. Furthermore, the elevated levels of ICAM- 1 and P- selectinwere found in the blood of ischemia/ reperfusion rats. Concluhon ICAM- 1 and P- selectin contribute tohepatic ischemia / reperfusion injury,and the P - selectin Mab can protect the liver against the injury.展开更多
AIM: To investigate a possible association between losartan and sirtuin 1(SIRT1) in reduced-size orthotopic liver transplantation(ROLT) in rats.METHODS: Livers of male Sprague-Dawley rats(200-250 g) were preserved in ...AIM: To investigate a possible association between losartan and sirtuin 1(SIRT1) in reduced-size orthotopic liver transplantation(ROLT) in rats.METHODS: Livers of male Sprague-Dawley rats(200-250 g) were preserved in University of Wisconsin preservation solution for 1 h at 4 ℃ prior to ROLT.In an additional group,an antagonist of angiotensin Ⅱ type 1 receptor(AT1R),losartan,was orally administered(5 mg/kg) 24 h and 1 h before the surgical procedure to both the donors and the recipients.Transaminase(as an indicator of liver injury),SIRT1 activity,and nicotinamide adenine dinucleotide(NAD+,a co-factor necessary for SIRT1 activity) levels were determined by biochemical methods.Protein expression of SIRT1,acetylated Fox O1(ac-Fox O1),NAMPT(the precursor of NAD+),heat shock proteins(HSP70,HO-1) expression,endoplasmic reticulum stress(GRP78,IRE1 a,p-e IF2) and apoptosis(caspase 12 and caspase 3) parameters were determined by Western blot.Possible alterations in protein expression of mitogen activated protein kinases(MAPK),such as p-p38 and p-ERK,were also evaluated.Furthermore,the SIRT3 protein expression and m RNA levels were examined.RESULTS: The present study demonstrated that losartan administration led to diminished liver injury when compared to ROLT group,as evidenced by the significant decreases in alanine aminotransferase(358.3 ± 133.44 vs 206 ± 33.61,P < 0.05) and aspartate aminotransferase levels(893.57 ± 397.69 vs 500.85 ± 118.07,P < 0.05).The lessened hepatic injury in case of losartan was associated with enhanced SIRT1 protein expression and activity(5.27 ± 0.32 vs 6.08 ± 0.30,P < 0.05).This was concomitant with increased levels of NAD+(0.87 ± 0.22 vs 1.195 ± 0.144,P < 0.05) the co-factor necessary for SIRT1 activity,as well as with decreases in ac-Fox O1 expression.Losartan treatment also provoked significant attenuation of endoplasmic reticulum stress parameters(GRP78,IRE1 a,p-e IF2) which was consistent with reduced levels of both caspase 12 and caspase 3.Furthermore,losartan administration 展开更多
目的探讨P38丝裂原活化蛋白激酶(p38MAPK)特异性活性抑制剂(FR167653)对大鼠肝脏缺血再灌注损伤保护作用的机理。方法利用封闭群SD大鼠肝脏部分缺血再灌注模型,将实验动物分假手术组、对照组、治疗组。分别在各个时间点按照相应程序采...目的探讨P38丝裂原活化蛋白激酶(p38MAPK)特异性活性抑制剂(FR167653)对大鼠肝脏缺血再灌注损伤保护作用的机理。方法利用封闭群SD大鼠肝脏部分缺血再灌注模型,将实验动物分假手术组、对照组、治疗组。分别在各个时间点按照相应程序采集标本检测肝组织髓过氧化物酶(MPO)活性、免疫组化检测黏附分子表达及肝组织诱生型一氧化氮合酶i NOS m RNA表达测定。结果肝脏缺血再灌注损伤后,对照组肝脏组织中MPO活性增高,而治疗组肝脏组织MPO活性明显低于对照组(t分别=3.30、3.10、4.10、2.90,P均<0.05);同时在治疗组中炎性趋化因子的表达也较对照组明显减低(t分别=3.70、4.30、4.60、4.90、3.50、4.00,P均<0.05)。免疫组化结果显示对照组肝组织内可见P-选择素(P-selectin)及细胞间黏附分子-1(ICAM-1)在肝组织中表达,而治疗组表达阳性率明显低于对照组;i NOS的m RNA表达水平在3 h和6 h时治疗组明显低于对照组(t分别=4.50、5.70,P均<0.05)。结论 FR167653能在多个环节、多个层次对缺血再灌注肝脏起着保护作用。展开更多
文摘目的:研究活血化瘀注射液I号(HHI-I)预处理与缺血预处理(ischemic preconditioning,IP)对大鼠肝脏缺血再灌注(ischemia and reperfusion, I/R)损伤的改善作用,并比较两者的作用效果．方法:健康♂SD大鼠80只,随机分为假手术对照组(Shaman)、缺血再灌注组(I／R组)、缺血预处理组(IP组)、HHI-I预处理组(HHI-I组),每组20只．建立大鼠部分肝缺血模型,各组在I／R后1,3,6,24 h分别取材,测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)的水平:取左肝测组织丙二醛(MDA)、超氧化物歧化酶(SOD)的含量．I／R后1 h RT- PCR检测肝组织中肿瘤坏死因子α(TNF-α)和细胞间黏附分子-1(ICAM-1)mRNA的表达, I／R后3 h行组织学观察．结果:I／R组、IP组、HHI-I组的ALT,AST, LDH活性、MDA值和TNF-α．ICAM-1 mRNA表达水平均明显高于Sham组．IP组、HHI-I组低于I／R组．HHI-I组所有时间点ALT,AST, LDH明显低于IP组(ALT:2378．8±303．4 nkat／L VS 2840．6±248．4 nkat／L;AST:2887．2±270．1 nkat／L vs 4567．6±275．1 nkat／L;LDH:10550．4±710．1 nkat／L vs 12164．1±735．1 nkat／L;P均<0．05)．HHI-I组MDA值明显低于IP组(17．35±1．39 nmol／g vs 21．66±1．84 nmol／g,P<0．05)．HHI-I组TNF-α,ICAM-1 mRNA表达水平低于IP组(TNF-α:0．54±0．06 vs 0．78±0．08; ICAM-1:0．43±0．03 vs 0．69±0．11,P均<0．01)．各组SOD值均低于Sham组(P<0．05)．IP组、HHI-I组均高于I／R组(P<0．05),HHI-I组SOD(1．3,6 h)明显高于IP组(136．00±12．50 nmol/gvs 124．70±9．32 nmol／g,P<0．05)．Sham组光镜下肝小叶结构正常;I/R组肝小叶结构紊乱,肝细胞水肿变性;IP组肝细胞水肿明显,部分肝细胞变性;HHI-I组肝小叶结构基本正常,肝细胞无明显水肿．结论:HHI-I预处理与IP均可改善I／R对肝脏造成的损伤,前者的效果优于后者．HHI-I的保护机制可能在于改善肝脏微循环,减轻组织缺氧状态,并通过抑制TNF-α和ICAM-1等细胞因子和细胞黏附�
文摘Objective To investigate the role of intercellular adhesion molecule-1 (ICAM-1) andP- selectin in hepatic ischemia/reperfusion injury. Methods The relationship between P- selectin as well asICAM- 1 and liver ischemia/reperfusion was studied, using a 60 minutes ischemia - reperfusion rat liver model.The animals were divided into three groups including: the sham group, the ischemic control group receiving onlythe normal saline and the treated group receiving anti-P-selection monoclonal antibody (Mab) at a dose of2mg/kg 15 minutes before reperfusion. The following indexes were analyzed: liver injury tests, liver histotogy,serum enzymes level (AST, ALT), and the levels of ICAM-1 and P-selectin in the blood of rats. ResultsSerum enzymes levels were significantly increased during sixty minutes of left lobar ischemia and reperfusion(saline control), and the increment was significantly inhibited with P- selectin Mab. Liver pathology observed bylight microscopy was greatly ameliorated by Mab. Furthermore, the elevated levels of ICAM- 1 and P- selectinwere found in the blood of ischemia/ reperfusion rats. Concluhon ICAM- 1 and P- selectin contribute tohepatic ischemia / reperfusion injury,and the P - selectin Mab can protect the liver against the injury.
基金Supported by Grants from Fondo de Investigaciones Sanitarias,No.FIS PI12/00519fellowship from Agència de Gestiód’Ajuts Universitaris i de Recerca,No.2012FI_B00382Generalitat de Catalunya,Barcelona,Catalonia,Spain(to Pantazi E)
文摘AIM: To investigate a possible association between losartan and sirtuin 1(SIRT1) in reduced-size orthotopic liver transplantation(ROLT) in rats.METHODS: Livers of male Sprague-Dawley rats(200-250 g) were preserved in University of Wisconsin preservation solution for 1 h at 4 ℃ prior to ROLT.In an additional group,an antagonist of angiotensin Ⅱ type 1 receptor(AT1R),losartan,was orally administered(5 mg/kg) 24 h and 1 h before the surgical procedure to both the donors and the recipients.Transaminase(as an indicator of liver injury),SIRT1 activity,and nicotinamide adenine dinucleotide(NAD+,a co-factor necessary for SIRT1 activity) levels were determined by biochemical methods.Protein expression of SIRT1,acetylated Fox O1(ac-Fox O1),NAMPT(the precursor of NAD+),heat shock proteins(HSP70,HO-1) expression,endoplasmic reticulum stress(GRP78,IRE1 a,p-e IF2) and apoptosis(caspase 12 and caspase 3) parameters were determined by Western blot.Possible alterations in protein expression of mitogen activated protein kinases(MAPK),such as p-p38 and p-ERK,were also evaluated.Furthermore,the SIRT3 protein expression and m RNA levels were examined.RESULTS: The present study demonstrated that losartan administration led to diminished liver injury when compared to ROLT group,as evidenced by the significant decreases in alanine aminotransferase(358.3 ± 133.44 vs 206 ± 33.61,P < 0.05) and aspartate aminotransferase levels(893.57 ± 397.69 vs 500.85 ± 118.07,P < 0.05).The lessened hepatic injury in case of losartan was associated with enhanced SIRT1 protein expression and activity(5.27 ± 0.32 vs 6.08 ± 0.30,P < 0.05).This was concomitant with increased levels of NAD+(0.87 ± 0.22 vs 1.195 ± 0.144,P < 0.05) the co-factor necessary for SIRT1 activity,as well as with decreases in ac-Fox O1 expression.Losartan treatment also provoked significant attenuation of endoplasmic reticulum stress parameters(GRP78,IRE1 a,p-e IF2) which was consistent with reduced levels of both caspase 12 and caspase 3.Furthermore,losartan administration
文摘目的探讨P38丝裂原活化蛋白激酶(p38MAPK)特异性活性抑制剂(FR167653)对大鼠肝脏缺血再灌注损伤保护作用的机理。方法利用封闭群SD大鼠肝脏部分缺血再灌注模型,将实验动物分假手术组、对照组、治疗组。分别在各个时间点按照相应程序采集标本检测肝组织髓过氧化物酶(MPO)活性、免疫组化检测黏附分子表达及肝组织诱生型一氧化氮合酶i NOS m RNA表达测定。结果肝脏缺血再灌注损伤后,对照组肝脏组织中MPO活性增高,而治疗组肝脏组织MPO活性明显低于对照组(t分别=3.30、3.10、4.10、2.90,P均<0.05);同时在治疗组中炎性趋化因子的表达也较对照组明显减低(t分别=3.70、4.30、4.60、4.90、3.50、4.00,P均<0.05)。免疫组化结果显示对照组肝组织内可见P-选择素(P-selectin)及细胞间黏附分子-1(ICAM-1)在肝组织中表达,而治疗组表达阳性率明显低于对照组;i NOS的m RNA表达水平在3 h和6 h时治疗组明显低于对照组(t分别=4.50、5.70,P均<0.05)。结论 FR167653能在多个环节、多个层次对缺血再灌注肝脏起着保护作用。
