大鼠部分肝缺血再灌注损伤后切除对肝再生的影响

目的 观察大鼠部分肝缺血再灌注损伤后切除对残肝再生的影响.方法 将75只健康雄性SD大鼠随机分为5组:肝脏左叶和中叶(约占全肝70%)切除组(Control组)、肝脏左叶和中叶缺血10min再灌注30min后切除组(I10R30组)、类推得到I60R30组、I90R30组、I90R60组.术后6、12、24h等时间点,测定再生肝重量(RLW);自动生化分析仪检测血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)含量;酶联免疫吸附试验(ELISA)检测血清肿瘤坏死因子(TNF)-α含量;通过免疫组织化学法检测残肝增殖细胞核抗原(Ki-67)表达.结果 术后12h,I60R30、I90R30和I90R60组RLW值分别为(1.80±0.03)%、(1.82±0.10)%、(1.87±0.05)%;Ki-67值分别为(58.35±2.18)%、(59.73±3.06)%、(62.65±2.24)%,均明显高于对照组(P<0.05).缺血再灌注干预各组ALT和AST明显高于对照组(P<0.05).术后6h和12h,I60R30、I90R30和I90R60组TNF-α明显高于对照组(P<0.05).结论 大鼠即将被切除的肝脏先缺血再灌注后切除,对残肝再生具有促进作用;诱导产生的TNF-α表达量增多是促进肝再生的原因之一.

Abstract：

Objective To investigate the effects of ischemia reperfusion injury before partial hepatectomy on liver regeneration in rats. Methods Seventy-five male healthy SD rats were randomly classified into 5 groups: group control, in which rats were only subjected to 70% hepatectomy; group I10R30, 70% liver hepatectomy after 10 min of ischemia and 30 min of reperfusion in the resected liver; By analogy, group I60R30, group I90R30 and group I90R60 were constructed. At 6th, 12th and 24th h after operation, RLW was determined; serum alanine aminotransferase (ALT) and aspartate transaminase (AST) activities were measured by using autoanalyzer; the levels of serum tumor necrosis factor (TNF)-α were determined by ELISA and the expression level of Ki-67 was detected by using immunohistochemical methods in the residual liver tissues. Results At 12th h after partial hepatectomy, the rate of RLW in group I60R30, group I90R30 and group I90R60 was (1.80±0.03)%, (1.82±0.10)%, (1.87±0.05)% respectively; the rate of Ki-67 was (58.35±2.18)%, (59.73±3.06)%, (62.65±2.24)% respectively, which was significantly higher than that in the group control (P<0.05). The levels of ALT and AST in rats with ischemia reperfusion injury were higher than in the group control (P<0.05). At 6th h and 12th h after operation, the expression levels of TNF-α in groups I60R30, I90R30 and I90R60 were significantly higher than those in the group control (P<0.05). Conclusion Ischemia reperfusion injury in the resected liver before partial hepatectomy could improve liver regeneration of the remnant liver in rats. The high expression of induced TNF-α may be one of the reasons.     

三氧化二砷对逆行隔离灌注大鼠肝脏毒性的观察

Objective To study As2O3toxicity on rat liver in a retrograde isolated hepatic perfusion model. Methods In this study 104 male Sprague-Dawley rats weighing between 300 and 400 g were used. Eight male SD rats were used for preoperatively normal control and the remaining rats were randomly divided into 4 subgroups receiving As2O3at dosage of 0 mg/kg,0.75 mg/kg, 1.5 mg/kg, 3 mg/kg respectively. Modified RIHP was used in which As2O3was infused through hepatic artery. Ringer's lactate was retrogradly infused through hepatic veins and the portal vein was used as the outflow tract. Hepatic function, pathology and liver enzymes were assessed at different time points. As2O3concentration was monitered during the perfusion in rats of subgroup C. Results Serum ALT and AST rose to the peak on the first day, returning to normal after 3 or 7 days in all four subgroups. There was no difference between the peak levels of serum ALT and AST between subgroup A and B. Differences in serum ALT、AST level between subgroup A and C, A and D, B and C, B and D, C and D were all statistically significant (FALT=40.811,P<0.01;FAST= 48.212,P <0.01). On day 7, ALT and AST in subgroup D were still statistically higher when compared with that of other subgroups and normal control (FALT=13.928, P<0.01;FAST=17.942, P<0.01), and the hepatic pathology showed necrosis of the hepatocyte. The peak levels of As2O3were 13.21±0.82(μg/ ml) and 0.09±0.008 (μg/ml)in rats liver and systemic circulation in subgroup C during isolated perfuision. There were significant differences between the peak levels of concentration of As2O3in rats liver and systemic circulation (t=35.758,P<0.01). Conclusions The hepatic toxicity is reversible caused by As2O3when given at a dosage of 1.5 mg/kg of As2O3in a murine model of RIHP.     

三氧化二砷对逆行隔离灌注大鼠肝脏毒性的观察

Objective To study As2O3toxicity on rat liver in a retrograde isolated hepatic perfusion model. Methods In this study 104 male Sprague-Dawley rats weighing between 300 and 400 g were used. Eight male SD rats were used for preoperatively normal control and the remaining rats were randomly divided into 4 subgroups receiving As2O3at dosage of 0 mg/kg,0.75 mg/kg, 1.5 mg/kg, 3 mg/kg respectively. Modified RIHP was used in which As2O3was infused through hepatic artery. Ringer's lactate was retrogradly infused through hepatic veins and the portal vein was used as the outflow tract. Hepatic function, pathology and liver enzymes were assessed at different time points. As2O3concentration was monitered during the perfusion in rats of subgroup C. Results Serum ALT and AST rose to the peak on the first day, returning to normal after 3 or 7 days in all four subgroups. There was no difference between the peak levels of serum ALT and AST between subgroup A and B. Differences in serum ALT、AST level between subgroup A and C, A and D, B and C, B and D, C and D were all statistically significant (FALT=40.811,P<0.01;FAST= 48.212,P <0.01). On day 7, ALT and AST in subgroup D were still statistically higher when compared with that of other subgroups and normal control (FALT=13.928, P<0.01;FAST=17.942, P<0.01), and the hepatic pathology showed necrosis of the hepatocyte. The peak levels of As2O3were 13.21±0.82(μg/ ml) and 0.09±0.008 (μg/ml)in rats liver and systemic circulation in subgroup C during isolated perfuision. There were significant differences between the peak levels of concentration of As2O3in rats liver and systemic circulation (t=35.758,P<0.01). Conclusions The hepatic toxicity is reversible caused by As2O3when given at a dosage of 1.5 mg/kg of As2O3in a murine model of RIHP.     

高渗氯化钠羟乙基淀粉40注射液高容量血液稀释对大鼠肝脏缺血再灌注损伤的影响

Objective To investigate the effects of hypervolemic hemodilution (HH) with hypertonic saline plus hetastarch solution 40 injectio on hepatic ischemia-reperfusion (I/R) injury in rats. Methods Thirty male Wistar rats weighing 300-350 g were randomly divided into 3 groups ( n = 10 each): group I sham operation (group S); group II I/R and group Ⅲ HH. Partial liver ischemia was produced by clamping hepatic portal vein and left arteria hepatica for 30 min with atraumatic mini-clamp, followed by 2 h of reperfusion in I/R group and HH group. In HH group the animals were infused hypertonic saline plus hetastarch solution 40 injectio 10 ml/kg through vena caudalis over 30 min and then hepatic I/R was performed IS min after the infusion.The animals were killed at 2 h of reperfusion. The left liver was removed and blood sample was taken from inferior caval vein for determination of (1) serum alanine amino transferase (ALT) and aspartate amino transferase (AST) activities; (2) superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content in the liver; ( 3 ) microscopic examination. Results The serum ALT and AST activities and MDA content in the liver were significantly higher, SOD activity in the liver significantly lower after hepatic I/R and pathological changes in the liver severer in group I/R and HH than in group S. The serum ALT and AST activities and MDA content in the liver were significantly lower, SOD activity in the liver significantly higher after hepatic I/R and pathological changes in the liver milder in group HH than in group I/R. Conclusion Hypervolemic hemodilution with hypertonic saline plus hetastarch solution 40 injectio can ameliorate hepatic I/R injury by decreasing oxygen free radical production in rats.     

高渗氯化钠羟乙基淀粉40注射液高容量血液稀释对大鼠肝脏缺血再灌注损伤的影响

Objective To investigate the effects of hypervolemic hemodilution (HH) with hypertonic saline plus hetastarch solution 40 injectio on hepatic ischemia-reperfusion (I/R) injury in rats. Methods Thirty male Wistar rats weighing 300-350 g were randomly divided into 3 groups ( n = 10 each): group I sham operation (group S); group II I/R and group Ⅲ HH. Partial liver ischemia was produced by clamping hepatic portal vein and left arteria hepatica for 30 min with atraumatic mini-clamp, followed by 2 h of reperfusion in I/R group and HH group. In HH group the animals were infused hypertonic saline plus hetastarch solution 40 injectio 10 ml/kg through vena caudalis over 30 min and then hepatic I/R was performed IS min after the infusion.The animals were killed at 2 h of reperfusion. The left liver was removed and blood sample was taken from inferior caval vein for determination of (1) serum alanine amino transferase (ALT) and aspartate amino transferase (AST) activities; (2) superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content in the liver; ( 3 ) microscopic examination. Results The serum ALT and AST activities and MDA content in the liver were significantly higher, SOD activity in the liver significantly lower after hepatic I/R and pathological changes in the liver severer in group I/R and HH than in group S. The serum ALT and AST activities and MDA content in the liver were significantly lower, SOD activity in the liver significantly higher after hepatic I/R and pathological changes in the liver milder in group HH than in group I/R. Conclusion Hypervolemic hemodilution with hypertonic saline plus hetastarch solution 40 injectio can ameliorate hepatic I/R injury by decreasing oxygen free radical production in rats.     

高渗氯化钠羟乙基淀粉40注射液高容量血液稀释对大鼠肝脏缺血再灌注损伤的影响

Objective To investigate the effects of hypervolemic hemodilution (HH) with hypertonic saline plus hetastarch solution 40 injectio on hepatic ischemia-reperfusion (I/R) injury in rats. Methods Thirty male Wistar rats weighing 300-350 g were randomly divided into 3 groups ( n = 10 each): group I sham operation (group S); group II I/R and group Ⅲ HH. Partial liver ischemia was produced by clamping hepatic portal vein and left arteria hepatica for 30 min with atraumatic mini-clamp, followed by 2 h of reperfusion in I/R group and HH group. In HH group the animals were infused hypertonic saline plus hetastarch solution 40 injectio 10 ml/kg through vena caudalis over 30 min and then hepatic I/R was performed IS min after the infusion.The animals were killed at 2 h of reperfusion. The left liver was removed and blood sample was taken from inferior caval vein for determination of (1) serum alanine amino transferase (ALT) and aspartate amino transferase (AST) activities; (2) superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content in the liver; ( 3 ) microscopic examination. Results The serum ALT and AST activities and MDA content in the liver were significantly higher, SOD activity in the liver significantly lower after hepatic I/R and pathological changes in the liver severer in group I/R and HH than in group S. The serum ALT and AST activities and MDA content in the liver were significantly lower, SOD activity in the liver significantly higher after hepatic I/R and pathological changes in the liver milder in group HH than in group I/R. Conclusion Hypervolemic hemodilution with hypertonic saline plus hetastarch solution 40 injectio can ameliorate hepatic I/R injury by decreasing oxygen free radical production in rats.     

高渗氯化钠羟乙基淀粉40注射液高容量血液稀释对大鼠肝脏缺血再灌注损伤的影响

Objective To investigate the effects of hypervolemic hemodilution (HH) with hypertonic saline plus hetastarch solution 40 injectio on hepatic ischemia-reperfusion (I/R) injury in rats. Methods Thirty male Wistar rats weighing 300-350 g were randomly divided into 3 groups ( n = 10 each): group I sham operation (group S); group II I/R and group Ⅲ HH. Partial liver ischemia was produced by clamping hepatic portal vein and left arteria hepatica for 30 min with atraumatic mini-clamp, followed by 2 h of reperfusion in I/R group and HH group. In HH group the animals were infused hypertonic saline plus hetastarch solution 40 injectio 10 ml/kg through vena caudalis over 30 min and then hepatic I/R was performed IS min after the infusion.The animals were killed at 2 h of reperfusion. The left liver was removed and blood sample was taken from inferior caval vein for determination of (1) serum alanine amino transferase (ALT) and aspartate amino transferase (AST) activities; (2) superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content in the liver; ( 3 ) microscopic examination. Results The serum ALT and AST activities and MDA content in the liver were significantly higher, SOD activity in the liver significantly lower after hepatic I/R and pathological changes in the liver severer in group I/R and HH than in group S. The serum ALT and AST activities and MDA content in the liver were significantly lower, SOD activity in the liver significantly higher after hepatic I/R and pathological changes in the liver milder in group HH than in group I/R. Conclusion Hypervolemic hemodilution with hypertonic saline plus hetastarch solution 40 injectio can ameliorate hepatic I/R injury by decreasing oxygen free radical production in rats.     

高渗氯化钠羟乙基淀粉40注射液高容量血液稀释对大鼠肝脏缺血再灌注损伤的影响

Objective To investigate the effects of hypervolemic hemodilution (HH) with hypertonic saline plus hetastarch solution 40 injectio on hepatic ischemia-reperfusion (I/R) injury in rats. Methods Thirty male Wistar rats weighing 300-350 g were randomly divided into 3 groups ( n = 10 each): group I sham operation (group S); group II I/R and group Ⅲ HH. Partial liver ischemia was produced by clamping hepatic portal vein and left arteria hepatica for 30 min with atraumatic mini-clamp, followed by 2 h of reperfusion in I/R group and HH group. In HH group the animals were infused hypertonic saline plus hetastarch solution 40 injectio 10 ml/kg through vena caudalis over 30 min and then hepatic I/R was performed IS min after the infusion.The animals were killed at 2 h of reperfusion. The left liver was removed and blood sample was taken from inferior caval vein for determination of (1) serum alanine amino transferase (ALT) and aspartate amino transferase (AST) activities; (2) superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content in the liver; ( 3 ) microscopic examination. Results The serum ALT and AST activities and MDA content in the liver were significantly higher, SOD activity in the liver significantly lower after hepatic I/R and pathological changes in the liver severer in group I/R and HH than in group S. The serum ALT and AST activities and MDA content in the liver were significantly lower, SOD activity in the liver significantly higher after hepatic I/R and pathological changes in the liver milder in group HH than in group I/R. Conclusion Hypervolemic hemodilution with hypertonic saline plus hetastarch solution 40 injectio can ameliorate hepatic I/R injury by decreasing oxygen free radical production in rats.     

肝脏缺血/再灌注后Toll样受体4表达情况及丙泊酚对其表达的影响

Objective To observe the changes in TLR4 expression during hepatic ischemia/reperfusion (I/R) and the effects of propofol on the expression of TLR4 induced by I/R injury in rats.Methods Fifty six male SD rats were randomly divided into min before isehemia in P groups,and the same volume of sodium lactate Ringer's solution was infused in sham group and I/R groups.Plasma ALT,AST,TNF-α levels were measured,and the expression of Tlr4 was detected 2,6,24 h after reperfusion.Results The levels of plasma ALT,AST and TNF-α were lower,and Tlr4 expression was weaker in P groups than those in I/R groups (P＜0.05).Conclusion Propofol decreases hepatic ischemia/reperfusion (I/R)-induced Tlr4 expression and exerts property of liver protection.     

脐带间充质干细胞旁分泌物质对暴发性肝衰竭大鼠肝功能及肝细胞增殖的影响

目的 探索人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,UC-MSCs)旁分泌物质对实验性暴发性肝衰竭大鼠的治疗作用,研究其对大鼠肝功能及肝细胞增殖的影响.方法 体外分离培养人脐带间充质于细胞,流式细胞仪检测UC-MSCs的表面标志,制备含有UC-MSCs旁分泌物质的条件培养基(MSC-CM),腹腔注射D-氨基半乳糖制备暴发性肝衰竭大鼠模型.实验分为3组:MSC-CM组、生理盐水(NS)组和促肝细胞生长素(PHGF)组.在模型制备后24 h从大鼠尾静脉分别注射三组治疗药物.每组8只大鼠治疗后12 h、24 h、36 h、60 h分别经内眦取血测定谷丙转氨酶(ALT)和总胆红素(TBIL)含量.每组另取5只大鼠在治疗后36 h取肝脏组织制备切片进行PCNA免疫组化染色,检测大鼠肝细胞增殖情况.观察并记录各组大鼠的生存状态及生存时间.结果 MSC-CM组及PHGF组大鼠治疗后24 h ALT及TBIL的含量均低于NS组(P＜0.05),MSC-CM组与PHGF组比较差异无统计学意义.大鼠治疗后36 h肝脏切片PCNA染色显示,MSC-CM组和PHGF组PCNA肝细胞阳性数显著高于NS组(P＜0.01),MSC-CM组与PHGF组比较差异无统计学意义.生存分析显示,MSC-CM组和PHGF组大鼠的生存率高于NS组(P=0.049),MSC-CM组与PHGF组比较差异无统计学意义.结论 人脐带间充质干细胞的旁分泌物质可以刺激暴发性肝衰竭大鼠肝细胞增殖,改善暴发性肝衰竭大鼠的肝功能,为暴发性肝衰竭的治疗提供了一种新途径.

Abstract：

Objective To investigate the therapeutic effect of human umbilical cord mesenchymal stem cell-paracrine substance on fulminant hepatic failure (FHF) rat, and to study the effect on liver function and hepatocyte proliferation. Methods Mesenchymal stem cells(MSCs)were separated from human umbilical cord, and surface makers of cells were detected by flow cytometry. Human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) was prepared. FHF rat model was induced by intraperitoneal injection of D-galactosamine and they were randomly diveded into three groups: MSC-CM group, NS group, PHGF group. 24 h later, 1 ml MSC-CM, 1 ml 0. 9% NaCl solution and lml PHGF solution was injected into the tail vein of MSC-CM, NS, and PHGF rats, respectively. In each group (n=8 per group), blood samples were collected at 12, 24, 36, and 60 h after treatment from inner canthus for analysis of blood ALT and TBIL levels. We used five rats per group for tissue collection after sacrifice at 36 h after treatment and 10 animals per group for survival analysis. PCNA immunohistochemical staining was used in the sections of liver tissue to detect hepatocyte proliferation. Results 24 h after treatment, the levels of ALT and TBIL in the MSC-CM and PHGF groups were lower than those in the NS group(P＜0. 05), but there was no significant difference between the MSC-CM and PHGF groups. There were more PCNA-positive hepatocytes in the MSC-CM and PHGF groups than in the NS group(P＜0.01), but there was no significant difference between MSC-CM and PHGF group. Survival analysis found that the survival rate of rats in the MSC-CM and PHGF groups was higher than that of rats in the NS group (P=0. 049), but there was no significant difference between the MSC-CM and PHGF group. Conclusions The paracrine substance of human umbilical cord mesenchymal stem cells can stimulate hepatocyte proliferation and improve liver function of FHF rats, potentially creating a new avenue for the treatment of FHF.     

RNA干扰Kupffer细胞肿瘤坏死因子-α减轻大鼠肝脏缺血再灌注损伤

目的 观察RNA干扰肝脏Kupffer细胞肿瘤坏死因子-α(TNF-α)对大鼠肝脏缺血再灌注损伤的保护作用.方法 构建针对大鼠TNF-α基因的短发夹状RNA(shRNA)真核表达载体.肝脏缺血再灌注损伤前48 h经门静脉注射磷酸盐缓冲液(PBS)、空载体或TNF-α shRNA.实验随机分为4组,假手术组、PBS组、空载体组和shRNA组.阻断大鼠70%入肝血流40 min,再灌注6 h检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、肝脏Kupffer细胞TNF-α mRNA、血清TNF-α、肝组织中丙二醛(MDA)以及超氧化物岐化酶(SOD)含量.结果 与PBS组和空载体组比较,shRNA组再灌注6 h后血清ALT和AST水平显著降低(P＜0.05),Kupffer细胞TNF-α mRNA水平、血清TNF-α水平(56.6±6.7 pg/ml比87.8±8.7 pg/ml和96.5±7.3 pg/ml,P＜0.05)、肝组织中MDA含量(93.4±13.3 nmol/mg比133.5±12.4 nmo1/mg和136.7±13.6 nmol/mg,P＜0.05)显著降低,SOD活性显著升高(22.4±4.6 U/mg比12.2±3.1 U/mg和11.4±2.9 U/mg,P＜0.05).结论 RNA干扰Kupffer细胞TNF-α基因的表达可以减轻大鼠肝脏缺血再灌注损伤.     

wortmannin对重症急性胰腺炎大鼠胰肝损伤的保护作用

目的 探讨wortmannin预处理对重症急性胰腺炎(SAP)大鼠肝损伤的保护作用及其可能机制。方法 健康成年SD大鼠54只，随机分为对照组（C组）、SAP组（P组）和SAP＋wortmannin组（PW组），每组18只。除C组以生理盐水代牛磺胆酸钠外，另两组逆行胆胰管注射50g/L牛磺胆酸钠制备SAP模型。各组分别于术后3，6，12h检测血清中TNF-α，ALT，AST水平及肝组织中NF-κB活性，观察肝组织及胰腺组织的病理变化。结果 P组血清中TNF-α，ALT，AST水平及肝组织中NF-κB活性（积分灰度）均显著高于C组（P<0.01）；胰腺、肝组织病理损伤随病情进展而逐渐加重。PW组较C组各项指标均升高，但较P组明显降低（P<0.01）；且胰腺、肝组织病理损伤亦较P组减轻。结论 wortmannin预处理对SAP大鼠胰肝损伤有一定的保护作用，其机制可能与其抑制了NF-κB的活化、减少TNF-α等多种炎症因子的释放有关。     

抑制Caspase-8基因表达对减轻大鼠肝脏缺血再灌注损伤的作用

目的 观察抑制半胱氨酸天冬氨酸特异性蛋白酶8(Caspase-8)基因的表达对减轻大鼠肝脏缺血再灌注损伤的作用.方法 构建针对大鼠Caspase-8基因的短发夹状RNA(shRNA)真核表达载体.将Lewis大鼠随机分为3组,每组8只.(1)假手术组:麻醉后,取腹部正中切口,缝合关腹;(2)磷酸盐缓冲液(PBS液)组:阻断肝门血流前48 h经门静脉注射PBS液1 ml,然后行肝脏缺血再灌注;(3)shRNA组:阻断肝门血流前48 h经门静脉注射Caspase-8 shRNA 50 μg(总体积为1 ml),然后行肝脏缺血再灌注.肝脏缺血再灌注的方法为阻断大鼠70%入肝血流40min.于再灌注6 h、12 h、24 h、3 d、5 d、7 d时检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平;检测肝组织中Caspase-8 mRNA的表达、细胞凋亡情况、丙二醛(MDA)以及超氧化物岐化酶(SOD)的含量.结果 与PBS液组比较,shRNA组再灌注6、12、24 h,血清中ALT和AST水平显著降低(P<0.05);肝组织中Caspase-8 mRNA水平、肝细胞凋亡指数(shRNA组和PBS液组分别为22.33%±4.28%和35.24%±2.33%)以及肝组织中MDA含量[shRNA组和PBS液组分别为(94.5±11.2)nmol/mg和(133.5±12.4)nmol/mg]均显著降低(P<0.05),而肝组织中SOD活性显著升高[shRNA组和PBS液组分别为(21.6±3.7)U/mg和(12.2±3.1)U/mg,P<0.05].结论 通过RNA干扰Caspase-8基因的表达可以抑制肝细胞凋亡的发生,减轻肝脏缺血再灌注损伤.     

丙泊酚对小鼠肝脏缺血再灌注损伤后氧化应激和炎症的影响

目的研究丙泊酚对小鼠肝脏缺血再灌注损伤的保护作用及其作用机制。方法 40只健康雄性C57小鼠随机分为假手术组(Sham)、肝脏缺血再灌注损伤组(IRI)、地塞米松组(DEM,5 mg/kg)和丙泊酚组(PPF,20 mg/kg),每组10只。用无创血管夹夹闭左、中叶肝蒂构建70%肝脏缺血再灌注损伤模型。再灌注6 h后,检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)和核因子κB(NF-κB)水平;评估肝脏过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)、超氧化物歧化酶(SOD)活性,MDA含量和病理形态;并检测血清和和肝脏肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)和白介素6(IL-6)的m RNA表达。结果 IRI组ALT、AST和LDH水平分别为(395.12±35.81)U/L、(155.37±19.22)U/L和(776.32±59.27)U/L而PPF组血清中ALT、AST和LDH表达水平分别为(144.81±14.22)U/L、(55.13±6.71)U/L和(439.27±45.12)U/L,较IRI组均明显降低(P0.05)。此外,与IRI组相比,PPF组NF-κB、MDA、TNF-α、IL-1β和IL-6的表达水平均明显降低(P0.05),肝脏病理损伤程度明显减轻,CAT、GPx和SOD活性均明显增高(P0.05)。结论丙泊酚对肝脏缺血再灌注损伤有保护作用,其作用机制与丙泊酚减轻炎症反应和抑制氧化应激相关。     

N-乙酰半胱氨酸对大鼠胆道梗阻肝功能损伤的保护作用及机制研究

目的 探讨N-乙酰半胱氨酸(N-acetylcysteine,NAC)对大鼠胆道梗阻所致肝功能损伤的保护作用及其机制。方法Wistar大鼠72只随机均分成3组：(1)胆道结扎+NAC组(DBL+NAC,n=24)：开腹结扎并切断胆总管,建模成功后经腹腔注射NAC（150 mg·kg-1·d-1）连续注射7 d;(2)胆道结扎组（DBL组,n=24）;(3)假手术组(SO组,n=24)：仅行开腹游离胆总管不予结扎和切断。建模成功后1、3、5、7d每组分别活杀6只大鼠,取静脉血及肝组织,检测肝功能、血浆肿瘤坏死因子α(TNF-α)在各时相点的变化并采用Griess法检测一氧化氮(NO)产生情况。结果 在DBL组、DBL+ NAC组谷-草转氨酶(AST)、血清谷丙转氨酶(ALT)、总胆红素(TBIL)、直接胆红素(DBIL)均随胆道梗阻时间延长而升高,但DBL组AST、ALT在各时间点均较DBL+NAC组明显升高(P＜0.05),而TBIL、DBIL在这两组间无明显差异(P＞0.05)。DBL组和DBL+ NAC组TNF-α、NO浓度变化也随梗阻时间延长而升高,但DBL组较DBL+ NAC组TNF-α、NO浓度升高更明显(P＜0.05)。结论N-乙酰半胱氨酸能有效改善胆道梗阻所致肝损害,并有可能是通过下调肝组织中TNF-α、NO的表达这一途径实现的。     

落新妇苷减轻大鼠肝脏缺血再灌注损伤的机制研究

目的 探讨落新妇苷对大鼠肝脏缺血再灌注损伤(HIRI)的保护作用.方法 SD大鼠,分为Sham组(假手术组)、HIRI组(缺血再灌注组)、落新妇苷组(低剂量组、中剂量组、高剂量组),建立大鼠肝脏缺血再灌注模型,分别于再灌注4h、8h、16h后取血液及肝脏标本.检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST);光镜下观察肝细胞显微结构变化;Western blot分析肝组织中HMGB1、TLR4、NF-kB、TNF-α的表达.结果 Sham组4h、8h、16 h的血清ALT为(58.11 ±4.81) U/L、(57.12±5.33) U/L、(57.63±4.54) U/L,HIRI组4h、8h、16 h的血清ALT为与(540.38±21.41) U/L、(831.21±20.11) U/L、(191.95±15.35) U/L,HIRI组与Sham组相比,血清ALT在4h、8h、16h均显著升高(P＜0.01).与HIRI组相比,不同剂量组的血清ALT水平在4h、8h、16h均显著降低,其中高剂量组4h、8h、16 h的ALT为(110.12±9.14) U/L、(168.56±11.52) U/L、(81.12±6.45) U/L,降低最明显(P＜0.01);Sham组4h、8h、16 h的血清AST为(167.16±11.55) U/L、(161.55±10.85) U/L、(168.41±11.26) U/L,HIRI组4h、8h、16h的血清AST为(978.83±19.82) U/L、(1514.36±25.22) U/L、(411.25±31.63) U/L,HIRI组与Sham组相比,血清AST在4h、8h、16 h均显著升高(P＜0.01),与HIRI组相比,不同剂量组的血清ALT水平在4h、8h、16 h均显著降低,其中高剂量组4h、8h、16 h的血清AST为(223.75±10.53) U/L、(412.14±23.59) U/L、(205.25±15.48) U/L,血清AST降低最明显(P＜0.01).光镜结果示药物组肝细胞损伤明显减轻.Western blot结果示:高剂量组HMGB1、TLR4蛋白的表达在4h、8h、16 h均较HIRI组显著下降(P＜0.05).高剂量组NF-kB、TNF-α蛋白的表达在术后8h、16 h较HIRI组显著下降(P＜0.05),但在术后4h与HIRI组比较差异无统计学意义(P＞0.05).结论 落新妇苷预处理可减轻大鼠HIRI,其作用机制可能与下调HMGB1/TLR4/NF-kB/TNF-d轴,进而抑制炎症有关.     

极化液对内毒素血症大鼠肝损伤的影响

目的研究葡萄糖-胰岛素-钾(极化液,GIK)对内毒素血症大鼠肝损伤的影响。方法雄性SD大鼠60只,体重200~250g,随机分为三组:对照组,脂多糖组(LPS组,LPS 8mg/kg),GIK组(LPS 8mg/kg+GIK 4ml·kg~(-1)·h~(-1))。采用全自动生化仪检测腹腔注射LPS后3d和5d大鼠血清丙氨酸氨基转移酶(ALT)及天门冬氨酸氨基转移酶(AST)含量,ELISA法检测三组大鼠肝组织匀浆TNF-α含量,并行HE染色观察肝组织病理变化,TUNEL免疫荧光检测肝实质细胞凋亡情况。结果与注射后3d比较,注射后5dLPS组大鼠血清ALT、AST、肝组织匀浆TNF-α含量明显升高,而GIK组大鼠血清ALT、AST、肝组织匀浆TNF-α含量明显下降(P0.05)。与对照组比较,注射后3dLPS组和GIK组大鼠血清ALT、AST、注射后5dLPS组大鼠血清ALT、AST明显升高(P0.05),注射后3、5dLPS组和GIK组大鼠肝组织匀浆TNF-α含量、肝损伤等级评分、肝细胞凋亡指数明显升高(P0.05)。与LPS组比较,注射后3dGIK组大鼠肝组织匀浆TNF-α含量、肝细胞凋亡指数明显降低(P0.05),注射后5dGIK组大鼠血清ALT、AST、肝组织匀浆TNF-α含量、肝损伤等级评分、肝细胞凋亡指数明显降低(P0.05)。结论腹腔注射LPS可引起大鼠肝损伤,导致肝功能改变及肝细胞破坏;GIK可减轻LPS诱导的大鼠肝损伤。     

门冬氨酸鸟氨酸对肝硬化肝癌术后肝功能的影响

目的：观察门冬氨酸鸟氨酸（LOLA）对肝癌合并肝硬化半肝切除术后肝功能的影响。方法：通过随机、对照的方法,将66例肝癌合并肝硬化行半肝切除患者分为2组,仅治疗组于术后1～7 d静脉应用LOLA。全部患者分别在术前及术后1、3、5、7、10、14 d抽外周静脉血检查谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TB）、直接胆红素（DB）。结果：治疗组术后1、3、5 d ALT明显低于对照组（P〈0.05）,术后3、5 d AST明显低于对照组（P〈0.05）。治疗组术后7 d ALT恢复正常的比率高于对照组。LOLA对术后TB、DB无影响。两组术后肝功能不全的发生率无差异（P=0.557）。结论：LOLA能促进肝癌肝硬化半肝切除术后肝脏功能的早期恢复。     

不同预处理对大鼠肝移植供肝保护作用的探讨

目的 通过应用缺血、加热等多种手段对大鼠供肝进行移植术前的预处理,比较各种预处理方法对大鼠肝移植供肝缺血再灌注损伤的保护作用. 方法 将SD大鼠50只,随机分为5组:半肝缺血预处理组、脾脏缺血预处理组、热休克预处理组、热休克+缺血预处理组及手术对照组,分别进行肝脏预处理后行模拟原位肝移植术,术后检测胆汁流量,术后24 h检测血清ALT,AST,ALP水平并观察肝脏形态学变化. 结果 半肝缺血预处理组、热休克预处理组移植后胆汁分泌量多于对照组,血清ALT,AST水平明显低于对照组(P<0.05);热休克+缺血预处理组的血清ALT水平低于对照组(P<0.05),胆汁分泌量及血清AST与对照组没有显著差异;脾脏缺血预处理组的胆汁分泌量多于对照组,血清ALT水平低于对照组(P<0.05). 结论 肝脏缺血预处理初始阶段保护作用最明显,将缺血及热休克预处理两种方法联合处理大鼠时,其保护作用弱于单独缺血或单独热休克的预处理方法;脾脏缺血预处理也具有保护肝脏的作用.