兔慢性肝损害急性肝衰竭动物模型的建立

Objective To establish an ideal animal model of acute-on-chronic liver failure (ACLF) in New Zealand white rabbits in order to provide a large animal model for further researches.Methods Totally 75 New Zealand rabbits were randomly divided into experimental group (n =70) and control group (n = 5 ). Rabbits in the experimental group were injected with CCl4 into the abdominal cavity twice every week and the doses of CCl4 were modified according to the index of liver function and the body weight, whereas those in the control group were treated with the same volume of saline. At the 10th week,48 New Zealand rabbits with hepatic fibrosis were randomly assigned to 4 groups and injected with CCl4 as before, D-Gal at a dose of 0. 65 g/kg body weight (BW), 0. 70 g/kg BW and 0. 75 g/kg BW, respectively. By observing and comparing the general state, survival time, biochemical indexes, and the histopathology, a method of establishing a stable animal model of acute hepatic failure was found. Results As compared with those in control group, the levels of ALT, AST, GGT, HA, LN and PC-Ⅲ in the experiment group were increased significantly, while the level of ALB was decreased at the end of 10 weeks. Typical features of hepatic fibrosis and the formation of pseudo-lobules were observed at the end of 10 weeks. After treatment with D-Gal, all rabbits in group Ⅰ survived with minimal changes in liver function tests. In group Ⅱ , there was a temporary hepatic injury, but no hepatic coma. Four of the 12 rabbits died (33. 3% ). In group Ⅲ , biochemical indexes changed obviously 12 h after the administration and hepatic injury reached its peak after 48 h. Ten of 12 rabbits were died of severe hepatic failure with a survival time of ( 53. 00 ± 25. 69) h. Histology of liver section revealed massive necrosis in nodules. In group Ⅳ , hepatic injury occurred early and severely. All the rabbits died of severe hepatic failure with a survival time of (32. 70 ± 17. 46) h. Conclusion The experimental model of ACLF could be established by injected with D-Gal in New Zealand rabbits with hepatic fibrosis, induced by CCl4 intraperitoneal injection for 10 weeks.The one induced by 0. 70 g/kg of D-galactosamine was more stable and showed similar clinical pathophysiological changes in human beings. So it can be a good experimental platform for studies of ACLF.     

兔慢性肝损害急性肝衰竭动物模型的建立

Objective To establish an ideal animal model of acute-on-chronic liver failure (ACLF) in New Zealand white rabbits in order to provide a large animal model for further researches.Methods Totally 75 New Zealand rabbits were randomly divided into experimental group (n =70) and control group (n = 5 ). Rabbits in the experimental group were injected with CCl4 into the abdominal cavity twice every week and the doses of CCl4 were modified according to the index of liver function and the body weight, whereas those in the control group were treated with the same volume of saline. At the 10th week,48 New Zealand rabbits with hepatic fibrosis were randomly assigned to 4 groups and injected with CCl4 as before, D-Gal at a dose of 0. 65 g/kg body weight (BW), 0. 70 g/kg BW and 0. 75 g/kg BW, respectively. By observing and comparing the general state, survival time, biochemical indexes, and the histopathology, a method of establishing a stable animal model of acute hepatic failure was found. Results As compared with those in control group, the levels of ALT, AST, GGT, HA, LN and PC-Ⅲ in the experiment group were increased significantly, while the level of ALB was decreased at the end of 10 weeks. Typical features of hepatic fibrosis and the formation of pseudo-lobules were observed at the end of 10 weeks. After treatment with D-Gal, all rabbits in group Ⅰ survived with minimal changes in liver function tests. In group Ⅱ , there was a temporary hepatic injury, but no hepatic coma. Four of the 12 rabbits died (33. 3% ). In group Ⅲ , biochemical indexes changed obviously 12 h after the administration and hepatic injury reached its peak after 48 h. Ten of 12 rabbits were died of severe hepatic failure with a survival time of ( 53. 00 ± 25. 69) h. Histology of liver section revealed massive necrosis in nodules. In group Ⅳ , hepatic injury occurred early and severely. All the rabbits died of severe hepatic failure with a survival time of (32. 70 ± 17. 46) h. Conclusion The experimental model of ACLF could be established by injected with D-Gal in New Zealand rabbits with hepatic fibrosis, induced by CCl4 intraperitoneal injection for 10 weeks.The one induced by 0. 70 g/kg of D-galactosamine was more stable and showed similar clinical pathophysiological changes in human beings. So it can be a good experimental platform for studies of ACLF.     

脾切除对肝纤维化大鼠肝脏PDGF-B表达及血清PDGF-BB水平的影响

目的 研究脾切除对肝纤维化大鼠肝脏PDGF-B表达和血清PDGF-BB水平的影响,探讨脾切除在肝纤维化治疗中的作用.方法 大鼠皮下注射CCl4,建立65只肝纤维化大鼠模型.于建模开始、肝纤维化期以及肝硬化期三组切除大鼠睥脏,并设立同期假手术组作为对照组.第一、二组及其对照组于建模后第10周,第三组及其对照组于建模后12周分别取大鼠肝脏和血液标本.用免疫组化SABC法测定肝脏PIGF-B的表达.HE染色检测肝纤维化,用双抗体夹心ELISA法测定血清PIGF-BB水平.结果 各切脾组PDGF-B阳性细胞A值(光密度值)明显低于其对照组;各切脾组血清PDGF-BB含量明显低于其对照组.根据HE染色,建模开始时切脾再行肝纤维化诱导,肝纤维化进程缓慢;建模第5周切脾后,肝脏病理状态比对照组明显缓解;肝硬化期切脾后,肝脏炎性改变和纤维化程度有一定减轻.结论 早期脾脏切除对实验性肝纤维化有一定延缓作用.脾切除后PDGF水平的下降可能是造成该延缓的机制之一.

Abstract：

Objective To explore the effects of splenectomy on hepatic fibrosis and on the expression of PDGF-B in the liver and PDGF-BB in the serum of rats with hepatic fibrosis. Methods By hypodermic injection CCl4, we established 65 rat models with hepatic fibrosis, splenectomies were performed in the three groups at different phases: before hypodermic injection CCl4 (A group), five weeks after hypodermic injection CCl4 (B group), and ten weeks hypodermic injection CCl4 (C group). The control groups were established at the same time, with samples of the livers and serum of the rats taken in different phases. The expressions of PDGF in the liver were detected by immunohistochemistry technique and the degree of hepatic fibrosis was detected by HE staining. The serum levels of PDGF-BB were analyzed by ELISA technique. Results Absorbance values of PDGF-B in the experimental group were significantly lower than the control groups (P＜0. 05). Serum levels of PDGF-BB of the rats after splenectomy were significantly lower than those in the control groups (P＜0.05). HE and Masson's staining showed that the progression of Hepatic fibrosis was slow in the A group. Hepatic pathologic state was significantly relieved in the B group and the inflammation and fibrosis was relieved in the C group. Conclusion Earlier period splenectomy could delay the proceeding of experimental hepatic fibrosis. After splenectomy the decline in the level of PDGF may be one of the mechanisms causing the delay.     

舒芬太尼延迟预处理对兔心肌缺血再灌注损伤的影响

Objective To investigate whether delayed preconditioning with sufentanil can protect against myocardial ischemia and reperfusion (IR) injury and the possible mechanism. Methods Thirty-six male New Zealand white rabbits 8-18 months old weighing 2.0-2.4 kg were used in this study. The animals were anesthetized with iv etomidate 2 mg/kg, tracheostomized and mechanically ventilated (VT 10-12 ml/kg, RR 20-25 bpm, FiO2 100%). Carotid artery and jugular vein were cannulated for BP and HR monitoring and fluid administration.Myocardial ischemia was induced by 30 min of occlusion of anterior descending branch of left coronary artery followed by 3 h of reperfusion. The animals were randomly assigned to one of 5 groups based on the sufentanil and/or naloxone the animals received 24 h before myocardial ischemia: group Ⅰ received normal saline (group IR,n=9); greup Ⅱ sufentanil 5 μg/kg (group S1, n=6); group Ⅲ sufentanil 10 μg/kg (group S2, n=9); group Ⅳ naloxone 10 mg/kg (group N, n =6) and group Ⅴ naloxone 10 mg/kg + sufentanil 5 μg/kg (group NS, n =6). Myocardial infarct size was measured using triphenyl tetrazolium (TIC) staining. Myocardial apoptosis was detected by TUNEL, and Bcl-2 and Bax protein expression was determined by immuno-histochemistry in group IR and S2. Results Sufentanil produced a dose-dependent reduction in infarct size as compared with group IR.Naloxone had no effect on infarct size but partially abolished sufentanil-induced cardio-protection. Sufentanil significantly decreased myocardial apoptosis and Bax protein expression but increased Bcl-2 protein expression as compared with group IR. Conclusion Sufentanil produces a delayed preconditioning against myocardial IR injury through activating opioid receptors and inhibiting cadiomyocyte apoptosis.     

硬膜外注射倍他米松-利多卡因混合液对兔神经根炎症及硬膜外粘连的影响

Objective To investigate the effects of epidural administration of a mixture of betamethasone and lidocaine on nerve root inflammation and epidural space adhesion in rabbits. Methods Twenty-four adult male New Zealand white rabbits weighing 2.0-2.1 kg, were randomly divided into 2 groups ( n = 12 each): control group and treatment group. A catheter was inserted into epidural space at L2,3 interspace. Twenty-four hour after epidural catheter placement, talcum powder 0.5 mg/kg was injected into epidural space to make the model of nerve root inflammation and epidural space adhesion. Three days later a mixture of lidocaine 2.5 mg/kg and betamethasone 0.25 mg/kg was injected via the epidural catheter in treatment group, while the equal volume of normal saline was given in control group. At 21 days after administration of lidocaine and betamethasone, the spinal cord was removed, and dura mater and nerve root were checked with naked eye, light microscope and electron microscope.The neutrophil count in the dura mater was determined. Results There was nerve root inflammation and epidural space adhesion in control group. The nerve root inflammation and epidural space adhesion was not observed in treatment group. The neutrophil count was reduced in treatment group (21 ± 12) compared with control group (250 ±43) ( P < 0.01) . Conclusion Epidural administration of a mixture of betamethasone and lidocaine can alleviate nerve root inflammation and epidural space adhesion.     

硬膜外注射倍他米松-利多卡因混合液对兔神经根炎症及硬膜外粘连的影响

Objective To investigate the effects of epidural administration of a mixture of betamethasone and lidocaine on nerve root inflammation and epidural space adhesion in rabbits. Methods Twenty-four adult male New Zealand white rabbits weighing 2.0-2.1 kg, were randomly divided into 2 groups ( n = 12 each): control group and treatment group. A catheter was inserted into epidural space at L2,3 interspace. Twenty-four hour after epidural catheter placement, talcum powder 0.5 mg/kg was injected into epidural space to make the model of nerve root inflammation and epidural space adhesion. Three days later a mixture of lidocaine 2.5 mg/kg and betamethasone 0.25 mg/kg was injected via the epidural catheter in treatment group, while the equal volume of normal saline was given in control group. At 21 days after administration of lidocaine and betamethasone, the spinal cord was removed, and dura mater and nerve root were checked with naked eye, light microscope and electron microscope.The neutrophil count in the dura mater was determined. Results There was nerve root inflammation and epidural space adhesion in control group. The nerve root inflammation and epidural space adhesion was not observed in treatment group. The neutrophil count was reduced in treatment group (21 ± 12) compared with control group (250 ±43) ( P < 0.01) . Conclusion Epidural administration of a mixture of betamethasone and lidocaine can alleviate nerve root inflammation and epidural space adhesion.     

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.     

家兔肝纤维化肝癌模型的研究

目的建立新西兰白兔的肝纤维化合并肝癌模型。方法32只新西兰白兔随机分为2组,每组16只。模型组给予CCl_4油溶液灌胃,每周2次,共8周,第6周末肝脏接种VX-2肿瘤;对照组陪同饲养。观察实验兔的一般情况、体重变化、肝功能变化情况,并行CT及B超影像学检查和病理学检查。结果模型组共存活11只。与对照组比较,模型组体重下降,血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、γ-谷氨酰转肽酶(γ-Glutamyl Transpeptidase,GGT)、总胆红素(Total Bilimbin,TBil)升高,血清白蛋白(albumin,ALB)和白球比(A/G)下降,CT和B超发现肝脏占位,病理检查证实模型组新西兰白兔出现肝纤维化肝癌。结论使用CCl_4灌胃法诱发新西兰白兔肝纤维化,并种植肝脏VX-2肿瘤,可以建立肝纤维化肝癌的模型。     

当归对兔肝纤维化肝癌介入治疗后肝纤维化的作用

目的 建立兔肝纤维化肝癌模型,观察当归对其行化疗栓塞(TACE)术后加重肝纤维化的预防作用,探讨当归预防肝纤维化的作用机制.方法 将55只新西兰大白兔随机表法分为对照组(n=10)、模型组(n=15)、介入组(n=15)和预防组(n=15).造模时给予腹腔注射纯CCl4剂量0.1 ml/kg,每周1次,共注射10周,同时给予5%的乙醇饮水,在第10周末于肝左叶种植VX2瘤.其中模型组:在第12周末经肝动脉注入生理盐水2ml;介入组:在行TACE术时注入平阳霉素1mg+碘油0.2ml;预防组:在行TACE术时注入平阳霉素1mg+碘油0.2ml+当归注射液6ml/kg.对照组:以相同剂量生理盐水注射.各组于第16周末检测透明质酸酶(HA)、层粘蛋白(LN)、Ⅲ型前胶原(PCⅢ)和Ⅳ型胶原(CⅣ),并进行肝组织苏木素咿红(HE)染色、胶原纤维(VG)染色和转化生长因子(TGF)-β1免疫组织化学染色.结果 模型组(7/10)和预防组(6/10)的肝纤维化病理分期主要处于肝纤维化Ⅱ期(P>0.05),介入组(7/9)主要处于肝纤维化Ⅲ期,与模型组(3/10)和预防组(4/10)分期比较差异有统计学意义(P<0.05);模型组、介入组和预防组血清HA、LN、PCⅢ和CⅣ明显高于对照组(P<0.05),预防组低于介入组(P<0.05),与模型组比较差异无统计学意义(P>0.05);对照组肝组织TGF-β1呈阴性表达,介入组肝组织TGF-β1表达明显增强,模型组和预防组TGF-β1表达较介入组减少.结论 用CCl4诱发兔肝纤维化后并种植VX2瘤,可以建立兔肝纤维化肝癌模型,行TACE术后可加重其肝纤维化的程度,当归对其有延缓作用,TGF-β1参与其调控机制.     

不同剂量的羟基磷灰石纳米微粒治疗兔肝VX2肿瘤的实验研究

目的研究不同剂量的羟基磷灰石纳米微粒(hydroxyapatitenanoparticle,nHAP)对兔肝VX2肿瘤的治疗作用。方法将60只新西兰白兔经肝内肿瘤种植2周后随机分成4组,每组15只。于兔上腹正中开腹,胃十二指肠动脉插管固定,经肝动脉灌注给药。A组为生理盐水(5ml)对照组;B组为低剂量nHAP组,肝动脉注入nHAP(0.5mg/kg);C组为中剂量nHAP组,肝动脉注入nHAP(5mg/kg);D组为高剂量nHAP组,肝动脉注入nHAP(50mg/kg)。4组实验动物分别于治疗前,治疗后7d、14d行SCT肝脏扫描,测量肿瘤的大小,并计算肿瘤生长率。治疗前,治疗后1d、7d行血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)检测。各组实验动物均记录治疗后的生存天数。结果治疗后7d、14d,中剂量nHAP组和高剂量nHAP组的肿瘤体积和生长率都小于生理盐水对照组,有显著性差异(P<0.05);低剂量nHAP组与对照组相比无统计学差异(P>0.05);中剂量nHAP组与同期高剂量nHAP组相比,无显著性差异(P>0.05)。术后1d,D组血清AST、ALT均升高,与对照组相比差异有统计学意义(P<0.05)。术后7d,D组AST、ALT降至正常。治疗后,A、B、C组AST、ALT均未升高。A、B、C、D组瘤兔治疗后生存期分别为(38.0±5.4)d、(39.5±6.6)d、(45.2±7.7)d、(45.6±8.2)d,C、D两组与对照组相比有显著性差异(P<0.05)。结论在一定剂量范围内,nHAP对兔肝VX2肿瘤的生长具有抑制作用,并能提高瘤兔的生存期,其肝功能损害是可逆的。     

四氯化碳、硫代乙酰胺和猪血清诱导大鼠肝纤维化模型的比较

目的：比较采用四氯化碳（CCl4）,硫代乙酰胺（TAA）及猪血清诱导3种方式制备大鼠肝纤维化模型的效果和特点。方法：将72只SD大鼠随机均分为：CCl4组,TAA组,猪血清组及对照组,分别每周2次皮下注射40% CCl4（0.5 mL/100 g）,0.03% TAA（200 mg/kg）,猪血清（0.5 mL/只）和生理盐水（0.1 mL/kg）。观察大鼠一般情况及体重变化;在造模第3,6,9周末,测定大鼠血清中的天冬氨酸氨基转移酶（AST）,谷氨酸氨基转移酶（ALT）,丙二醛（MDA）,透明质酸（HA）水平;取肝组织进行HE染色,观察肝组织结构变化,并对肝纤维化程度进行分级和评分。结果：CCl4,TAA组大鼠体重增加量均明显低于对照组（均P<0.05）,而猪血清组与对照组间体重增加量差异无统计学意义（P>0.05）;TAA组3个时间点ALT浓度均明显升高（均P<0.05）,而CCl4组和猪血清组ALT无明显升高。CCl4组AST浓度在第3周明显升高（P<0.05）,但在第6,9周有所下降,TAA组3个时间点AST浓度均明显升高（均P<0.05）,猪血清组AST浓度无明显升高。3个实验组MDA和HA浓度在3个时点间均有所升高（均P<0.05）,两者均在TAA组升高最明显;3个实验组第9周均可见不同程度的肝细胞颗粒样变,汇管区纤维组织异常增生;与对照组比较,3个实验组的肝纤维化分级和SSS计分差异均具有统计学意义（均P<0.05）,第9周时,TAA组的SSS计分高于CCl4组（P<0.05）,而CCl4组的SSS评分高于猪血清组（P<0.05）。结论：3种方法均可诱导大鼠肝纤维化模型,TAA效果略优于CCl4。猪血清法造模对动物整体损伤较轻微。     

血管钠肽抑制肝纤维化的实验研究

目的探讨人工合成的钠尿肽-血管钠肽(VNP)对肝纤维化的抑制作用。方法雄性Balb/c小鼠,随机分为玉米油对照组(对照组:腹腔注射玉米油1mL/kg,每周2次,持续12周),肝纤维化模型组(模型组:腹腔注射CCl41mL/kg,每周2次,持续12周,最后6周静脉注射生理盐水1mL/kg.d)和肝纤维化模型+VNP治疗组(VNP治疗组:腹腔注射CCl41mL/kg,每周2次,持续12周,最后6周静脉注射VNP50μg/kg.d)。于末次注射后3d取各组小鼠肝脏标本,观察肝脏病变程度和纤维化水平。体外培养鼠肝星状细胞株(HSC-T6),并加以不同浓度的VNP处理,分别采用[3H]-胸苷和[3H]-脯氨酸掺入的方法检测细胞DNA和胶原的合成水平。结果和对照组比较,模型组出现显著肝脏损伤和胶原沉积,而VNP治疗能明显减轻CCl4诱导的肝脏损伤和胶原沉积;对照组,模型组,VNP治疗组肝脏胶原浓度分别为(43.6±6.3)μg/mg,(93.5±7.2)μg/mg,(62.2±5.1)μg/mg,差异有统计学意义(P<0.05);VNP(10-7mol/L)处理后,HSC-T6的[3H]-胸苷和[3H]-脯氨酸掺入量分别...     

控制性低中心静脉压对兔肝缺血再灌注损伤的影响

目的 评价控制性低CVP对兔肝缺血再灌注损伤的影响.方法 新西兰大白兔32只,随机分为4组(n=8):假手术组(S组)、控制性低CVP组(L组)、肝缺血再灌注组(IR组)和控制性低CVP下肝缺血再灌注组(LIR组).L组静脉输注硝酸甘油10～30μg·kg~(-1)·min~(-1)和多巴胺30～40μg·kg~(-1)·min~(-1),在5 min内使CVP降至4～5 cm H_2O且维持MAP≥90 mm Hg,持续至再灌注6 h.IR组采用夹闭肝门30 min后再开放的方法建立肝缺血再灌注模型.LIR组在控制性低CVP模型制备成功后立即进行肝缺血再灌注.分别于实施控制性低CVP前(T_0,基础状态)、再灌注即刻(T_1)、30 min(T_2)、1 h(T_3)、2 h(T_4)、4 h(T_5)、6 h(T_6)时采用彩色超声多普勒诊断仪测定门静脉、肝动脉和肝静脉的血流速度,同时采集动脉血样,测定血浆AST和ALT的活性.于再灌注6 h时,取肝组织,电镜下观察细胞超微结构.结果 与S组比较,L组各时点门静脉、肝动脉、肝静脉的血流速度、血浆AST和ALT的活性差异无统计学意义(P＞0.05),IR组T_(1～5)时肝动脉血流速度减慢,T_(5,6)时肝静脉血流速度增快,血浆AST和ALT的活性升高,LIR组T_(1～6)时肝静脉血流速度度增快,T_(1～6)时血浆AST和ALT的活性升高(P＜0.05);与IR组比较,LIR组T_(1,2)时肝动脉血流速度增快,T_1～6时肝静脉血流速度增快,T_(1,4～6)时血浆ALT和AST的活性降低(P＜0.05).与IR组比较,LIR组肝细胞线粒体及窦周间隙面微绒毛的肿胀程度减轻,肝血寞窦壁覆盖完整.结论 控制性低CVP可减轻兔肝缺血再灌注损伤,其机制可能与增加再灌注期间肝血流量,减轻肝细胞及肝血窦损伤,从而改善肝灌注有关.     

肝纤维化兔最低肺泡有效浓度

背景七氟醚广泛应用于外科手术患者,其可能会影响肝功能和肝血流。然而,肝纤维化对七氟醚最低肺泡有效浓度（minimum alveolar concentration,MAC）的影响仍不清楚。因此我们设计了这个研究,以测定肝纤维化兔的MAC。方法30只雄性新西兰白兔,体重约2．5kg,随机分为两组：肝纤维化组（n=20）和正常对照组（n=10）。肝纤维化组兔用50％的四氯化碳处理12周,以诱导肝纤维化。麻醉前测定血清总蛋白、白蛋白、球蛋白、总胆汁酸、丙氨酸氨基转移酶、门冬氨酸氨基转移酶、碱性磷酸酶、γ-谷氨酰转肽酶、总胆红素、直接胆红素和间接胆红素浓度。两组麻醉诱导和维持均用七氟醚。实验中运用标准的钳尾技术来测定兔自主呼吸时的七氟醚MAC。麻醉后处死兔子,取出肝脏做病理检查。结果肝纤维化组经过12周四氯化碳处理后,20只兔子中有14只存活;对照组10只兔子有9只存活。肝纤维化组存活的14只兔子均发展为中至重度的肝纤维化。其中14只兔子中有3只兔子由于其他疾病或对疼痛刺激无反应而被排除。与对照组相比,肝纤维化组的球蛋白、门冬氨酸氨基转移酶和γ-谷氨酰转肽酶的水平显著增高。然而,白蛋白和碱性磷酸酶的水平明显低于对照组。七氟醚麻醉期间,两组动物平均动脉血压、心率、呼气末二氧化碳和温度均稳定。肝纤维化组的七氟醚MAC显著低于对照组（3．52％和4．10％,P=0．018）。结论肝纤维化兔的七氟醚MAC显著降低。     

亚甲蓝对兔肝缺血再灌注损伤的影响

目的 探讨亚甲蓝对兔肝缺血再灌注损伤的影响.方法 健康成年新西兰大白兔24只,雌雄不拘,体重2.0～2.3 kg,随机分为3组(n=8):假手术组(S组)、肝缺血再灌注组(I/R组)和亚甲蓝组(MB组).I/R组及MB组采用夹闭肝左外叶、中叶、右中叶及方形叶肝动脉分支40min再灌注60 min的方法制备肝缺血再灌注模型,S组仅游离相应血管.MB组于再灌注前20 min经耳缘静脉注射亚甲蓝5 mg/kg(用生理盐水稀释至5 ml),S组及I/R组给予等容量生理盐水.于缺血前即刻(T1)、缺血20 min(T2)、加min(T3)、再灌注1 min(T4)、再灌注5 min(T5)、30 min(T6)、60 min(T7)时记录MAP和HR.于T1,5-7时取股动脉血样1 ml,测定血清TNF-α及IL-6的浓度.分别于T1,6,7时取股动脉血样1.5 ml,测定血浆ALT及AST的活性.于T7时测定肝左叶组织SOD活性及MDA含量,光镜下观察肝组织病理学结果.结果 与S组比较,I/R组T4-7,时MAP降低,T7时HR降低,肝组织SOD活性降低,MDA含量升高,I/R组及MB组T3-5时血清TNF-α和IL-6浓度升高,T6,7时血浆ALT和AST活性升高(P<0.05或0.01);与I/R组比较,MB组T4-7时MAP升高,肝组织SOD活性升高,MDA含量降低,T3-5时血清TNF-α和IL-6浓度降低,T6,7时血浆ALT和AST活性降低(P<0.05或0.01).MB组肝组织损伤较I/R组减轻.结论 亚甲蓝可维持血液动力学稳定,减轻兔肝缺血再灌注损伤.     

钆塞酸二钠增强MRI对奥沙利钳诱导C57BL/6小鼠肝功能损伤的评估价值

目的探讨钆塞酸二钠增强MRI对奥沙利钳诱导C57BV6小鼠肝功能损伤的评估价值。方法雄性无特定病原体C57BI>6小鼠40只,6周龄,体质量19-23 g0使用随机数字随机分成:对照组、实验A组、实验B组和实验C组,各10只。对照组腹腔注射生理盐水,实验组每周2次腹腔注射奥沙利钳,实验A组、B组、C组分别连续注射2、4、6周。测量增强MRI肝月旦特异期T1弛豫时间、首过快速上升期中的增强斜率百分比(ESP)O检测血清胆红素和白蛋白,计算白蛋白胆红素(ALBI)评分。病理组织染色观察肝组织损伤和纤维化。受试者工作特征(ROC)曲线评价ALBI评分、肝胆特异期T1弛豫时间、ESP诊断肝功能损伤的价值。结果实验组(A组、B组和C组)中16只小鼠纳入变性组(不伴有纤维化的肝细胞变性),14只纳入肝纤维化组。肝纤维化组肝胆特异期T1弛豫时间高于对照组和变性组,差异有统计学意义(均P<0.05)。对照组、变性组、肝纤维化组ESP呈升高趋势,差异有统计学意义(均P<0.05)。与对照组比较,变性组、肝纤维化组ALBI评分均降低,差异有统计学意义(均P<0.05)。肝纤维化组小鼠,ALBI评分、肝胆特异期T\弛豫时间和ESP诊断肝功能损伤的ROC曲线下面积分别为0.734.0.962和0.989。结论钆塞酸二钠增强MRI肝胆特异期T1弛豫时间和ESP可有效评估奥沙利钳诱导的C57BL/6小鼠肝功能损伤。     

经皮瘤内注射艾迪注射液治疗兔移植性VX2肝肿瘤

目的探讨超声引导下复方中药艾迪注射液瘤内注射治疗兔移植性VX2肝肿瘤的作用效果及机制。方法建立新西兰大白兔移植性肝VX2肿瘤模型28只,随机分为3组,即艾迪注射液组（12只）、无水乙醇组（PEI组,8只）、生理盐水组（8只）。全部动物在接种2周后每隔3天向肿瘤内注入治疗药物,注射4次后处死动物。于术前术后兔耳缘静脉抽血行血生化检查,同时行64排螺旋CT扫描测量并计算肿瘤体积、肿瘤增长率及肿瘤坏死率,以及进行组织细胞学与细胞超微结构的观察。结果各实验组病灶均进行性增大,但艾迪注射液组肿瘤增长率及坏死率最小,与生理盐水组比较差异均有统计学意义（P〈0.05）,与无水乙醇组比较差异无统计学意义;TUNEL法检测显示艾迪注射液组凋亡指数明显高于无水乙醇组及生理盐水组（P〈0.05）;艾迪注射液组肝肾功能及白细胞计数治疗前后均无明显差异,组内及组间比较差异无统计学意义（P〉0.05）。结论艾迪注射液可诱导肿瘤细胞凋亡,对肿瘤细胞具有细胞毒性作用。