三氧化二砷对逆行隔离灌注大鼠肝脏毒性的观察

目的 观察不同剂量三氧化二砷(As2O3)在大鼠逆行隔离灌注(RIHP)模型中对肝脏毒性作用.方法 104只体重300～400 g雄性SD大鼠中8只为术前正常对照组,其余大鼠分A、B、C、D 4组,每组24只.A组灌注不含As2O3的乳酸林格氏液,B、C、D组灌注As2O3的剂量分别为0.75 mg/kg、1.5 mg/kg、3 mg/kg.实验组均采用RIHP,即经肝动脉灌注As2O3溶液,经肝静脉逆行灌注乳酸林格氏液,门静脉为流出道;灌注时间为15 min,速度1 ml/min.对照观察实验组术后1、2、37 d各时点肝酶学变化、肝组织学改变和术后生存情况;检测C组大鼠术中和术后第1天的肝循环和体循环中As2O3浓度.结果 实验组ALT、AST峰值均出现在术后第1天,至术后3～7 d恢复正常.A和B组ALT、AST峰值间差异无统计学意义,A和C,A和D,B和C,B和D、C和D各组间ALT、AST峰值差异均有统计学意义(FALT=40.811,FAST=48.212,均P<0.01).D组术后第7天时仍与术前正常对照组及A、B、C各组间差异均有统计学意义(FALT=13.928,FAST=17.942,均P<0.01),且术后肝组织出现片状坏死等较严重的损害.肝循环和体循环As2O3血药浓度峰值分别为(13.21±0.82)μg/ml和(0.09±0.008)μg/ml,二者比较差异有统计学意义(t=35.758,P<0.01).结论 在人鼠RIHP模型中,当As2O3灌注剂量不超过1.5 mg/kg时肝脏遭受的损害是可逆的.     

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.     

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.     

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.     

HTK液逆行灌注可减轻肝脏隔离化疗对大鼠的损伤

目的:建立大鼠肝脏隔离灌注化疗模型;探讨在大剂量化疗药物肝脏隔离灌注过程中,HTK液经肝静脉逆行灌注对肝脏的保护及减少全身泄漏作用。方法:将75只体重300～350g的雄性SD大鼠随机分为3组,每组25只;手术建立大鼠隔离灌注化疗模型。A组经肝动脉灌注含30mg/L三氧化二砷的林格液,门静脉灌注林格液,肝静脉为流出道。B组经肝静脉灌注林格液,门静脉为流出道,余同A组。C组经肝静脉灌注4℃组氨酸-色氨酸-酮戊二酸(Histidine-Tryptophan-Ketoglutarat,HTK)液,余同B组。各组于术后1、2、3、7d各随机处死大鼠5只,进行血清肝功能及肝组织病理学检查。A组和B组大鼠术中检测体循环和肝循环中的血药浓度。结果:各组动物血清ALT和AST峰值均出现在术后第1天,其后开始下降,至术后7d恢复正常。术后第1、2、3d,3组间的血清ALT、AST均数均有显著性差异(P0.05);光镜下观察肝组织,进一步证实了上述结果。A组与B组间体循环血药浓度差异有统计学意义(P0.05),而两组间的肝循环血药浓度相类似;表明逆行隔离灌注的体循环泄漏率较顺行隔离灌注组为低。结论:低温HTK液逆行灌注可显著减轻肝脏隔离化疗对大鼠的损伤,提高手术的安全性。     

逆行隔离灌注化疗可减轻对大鼠肝脏的毒性及药物的泄漏

目的 采用大鼠肝脏隔离灌注模型探讨逆行隔离灌注(RIHP)较顺行隔离灌注(IHP)能否减少正常肝组织损伤及化疗药物外周泄漏率.方法 将90只体重300～350 g雄性SD大鼠随机分为A、B、C三组,每组30只:A组为空白对照组,经肝动脉及门静脉灌注乳酸林格液,以下腔静脉为灌注液流出道;B组行IHP,经肝动脉灌注含有350 mg/kg的氟尿嘧啶(5-Fu),门静脉灌注乳酸林格液,以下腔静脉为灌注液流出道;C组行RIHP,经肝动脉灌注含有350 mg/kg的氟尿嘧啶(5-Fu),经下腔静脉灌注乳酸林格液,以门静脉为灌注液流出道.术后1、3、5、7 d分别行血清ALT测定及肝组织病理学检查;高效液相色谱分析仪检测B、C组术中外周血药浓度.结果 三组术后3 d存活率分别为90.0%、86.7%和90.0%,三者差异无统计学意义.三组血清ALT均在术后第一天达到峰值,A组为(481.6±207.6)μmol/L;B组为(1641.6±658.0)μmol/L;C组为(913.0±353.5)μmol/L.B、C组均显著高于A组(P＜0.05);B组显著高于C组(P＜0.05).B组与C组术中外周血药浓度峰值分别为(131.2±29.4)μg/ml和(65.3±28.4)μg/ml.两组外周浓度有显著性差异(P＜0.05).A组术后肝脏病理改变较轻,术后7 d基本恢复正常;B组术后肝脏病理学改变相对严重,术后7 d局部仍可见坏死灶;C组术后肝脏病理改变后较A组严重,但较B组轻,术后7 d基本恢复正常.结论 RIHP较之IHP能够显著减轻化疗药物对正常肝组织的毒副作用和药物的外周泄漏,有望成为一种对肝癌更加有效安全的区域化疗方法.     

逆行隔离灌注化疗可减轻对大鼠肝脏的毒性及药物的泄漏

Objective The retrograde isolated hepatic perfusion (RIHP) model was used to compare with the isolated hepatic perfusion (IHP) model in reducing the rate of normal hepatic tissue toxicity and peripheral drug leakage during chemotherapy in rat liver. Methods A total of 90 male Sprague-Dawley rats weighing 300-350 g were randomized into 3 groups with 30 rats in each. Group A: perfusion with Lactated Ringer'S Solution through arteria hepatica (RA) and portal vein (PV),the inferior vena cava was used as an outflow tract of perfusate. Group B: For isolated hepatic perfusion (IHP), Fluorouracil (5-FU) was added into the perfusate at a dose of 350mg/kg and introduced in to the liver through arteria hepatica, portal vein was perfused by Lactated Ringer'S Solution, and the inferior vena cava was used as an outflow tract of perfusate. Group C: by using retrograde isolated hepatic perfusion (RIHP), the solution which contains 350 mg/kg Fluorouracil (5-FU) was also introduced through arteria hepatica, the inferior vena cava was introduced with Lactated Ringer'S Solution;the portal vein was used as an outflow tract of the perfusate. On day 1, 3, 5 and 7 after the perfusion in all groups, blood serum ALT test and liver histopathology test were performed. The peripheral blood drug levels were measured with high performance liquid chromatographic(HPLC) system in group B and group C. Results The survival rate was 90%, 86.7% and 90% in group A, B and C,respectively. No statistically significant difference was observed in the survival rate among the 3groups. In all the three groups, serum ALT levels were the highest on the first day after IHP: (481.6±207.6)μmol/LingroupA;(1641. 6±658.0) μmol/LingroupBand( 913. 0±353. 5)μmol/Lin group C. Significant higher serum ALT levels were observed by comparing group B and C with A(P＜0. 05). Meanwhile, the serum ALT levels were significantly higher in group B than in group C (P＜0.05). The peaks of peripheral blood drug concentration during the perfusion were 131.2±29.4μg/ml in group B and 65.3±28. 4μg/ml in group C. Significant difference was observed (P＜0. 05). Liver biopsies of group A showed mild changes on the first day after IHP and returned to normal after 7 days. Group B showed severe changes on the first day after IHP and local necrosis still existed after 7 days. Group C showed moderate changes as compared with group B on the first day after IHP and also returned to normal after 7 days. Conclusion Retrograde isolated hepatic perfusion (RIHP) can reduce the liver toxicity compared to isolated hepatic perfusion (IHP). Hopefully, RIHP will be considered as a safer way in regional chemotherapy in liver cancer.     

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.     

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.