Rifaximin on epigenetics and autophagy in animal model of hepatocellular carcinoma secondary to metabolic-dysfunction associated steatotic liver disease

BACKGROUND Prevalence of hepatocellular carcinoma(HCC)is increasing,especially in patients with metabolic dysfunctionassociated steatotic liver disease(MASLD).AIM To investigate rifaximin(RIF)effects on epigenetic/autophagy markers in animals.METHODS Adult Sprague-Dawley rats were randomly assigned(n=8,each)and treated from 5-16 wk:Control[standard diet,water plus gavage with vehicle(Veh)],HCC[high-fat choline deficient diet(HFCD),diethylnitrosamine(DEN)in drinking water and Veh gavage],and RIF[HFCD,DEN and RIF(50 mg/kg/d)gavage].Gene expression of epigenetic/autophagy markers and circulating miRNAs were obtained.RESULTS All HCC and RIF animals developed metabolic-dysfunction associated steatohepatitis fibrosis,and cirrhosis,but three RIF-group did not develop HCC.Comparing animals who developed HCC with those who did not,miR-122,miR-34a,tubulin alpha-1c(Tuba-1c),metalloproteinases-2(Mmp2),and metalloproteinases-9(Mmp9)were significantly higher in the HCC-group.The opposite occurred with Becn1,coactivator associated arginine methyltransferase-1(Carm1),enhancer of zeste homolog-2(Ezh2),autophagy-related factor LC3A/B(Map1 Lc3b),and p62/sequestosome-1(p62/SQSTM1)-protein.Comparing with controls,Map1 Lc3b,Becn1 and Ezh2 were lower in HCC and RIF-groups(P<0.05).Carm1 was lower in HCC compared to RIF(P<0.05).Hepatic expression of Mmp9 was higher in HCC in relation to the control;the opposite was observed for p62/Sqstm1(P<0.05).Expression of p62/SQSTM1 protein was lower in the RIF-group compared to the control(P=0.024).There was no difference among groups for Tuba-1c,Aldolase-B,alpha-fetoprotein,and Mmp2(P>0.05).miR-122 was higher in HCC,and miR-34a in RIF compared to controls(P<0.05).miR-26b was lower in HCC compared to RIF,and the inverse was observed for miR-224(P<0.05).There was no difference among groups regarding miR-33a,miR-143,miR-155,miR-375 and miR-21(P>0.05).CONCLUSION RIF might have a possible beneficial effect on preventing/delaying liver carcinogenesis through epigenetic modulation in a rat model of MASLD-HCC.

EVALUATION OF CIRRHOTIC LIVER WITH PERFUSION-WEIGHTED MAGNETIC RESONANCE IMAGING:A PRELIMINARY EXPERIMENTAL STUDY IN ANIMAL MODELS WITH HALF-LIVER CIRRHOSIS

Objective To investigate the role of peffusion-weighted magnetic resonance imaging （MRI） in evaluation of cirrhotic fiver. Methods With a 4F catheter, 1% diluted carbon tetrachloride （ 1 ml/kg） was selectively injected into fight or left hepatic artery of 12 dogs fortnightly. The half fiver into which carbon tetrachloride was injected was called as study side （SS）, while the other half fiver without carbon tetrachloride injection was called as study control side （SCS）. Conventional and peffusion-weighted MRI were performed in every 4 weeks. Via a 4F catheter, 5ml gadolinium diethylentriamine pentaaceti acid （Gd-DTPA） dilution was injected into superior mesenteric artery at the 5th scan. The signal intensity-thne curves of SS, SCS, and portal vein were completed in MR workstation. The maximal relative signal increase （ MRSI）, peak time （ tp）, and slope of the curves were measured. Results On conventional MR images, no abnormalities of externality and signal intensity were observed in both SS and SCS of fiver at each stage. The mean tp, MP, SI, and slope of intensity-time curves in normal fiver were 10. 56 seconds, 1.01, and 10. 23 arbitrary unit （au）/s, respectively. Three parameters of curves didn＇t show obvious change in SCS of fiver at every stage. Abnormal perfusion curves occurred in SS of fiver at the 12th week after the 1st injection. The abnormality of perfusion curve in SS was more and more serious as the times of injection increased. The mean tp, IVlRSI, and slope intensity-time curves in SS of fiver were 19.45 seconds, 0. 43, and 3. 60 au/s respectively at the 24th week. Conclusion Perfusion-weighted imaging can potentially provide information about portal peffusion of hepatic parenchyma, and to some degree, reflect the severity of cirrhosis.

Early changes of hepatic hemodynamics measured by functional CT perfusion in a rabbit model of liver tumor

BACKGROUND:Early detection and treatment of hepatocellular carcinoma is crucial to improving the patients’ survival.The hemodynamic changes caused by tumors can be serially measured using CT perfusion.In this study,we used a CT perfusion technique to demonstrate the changes of hepatic hemodynamics in early tumor growth,as a proof-of-concept study for human early hepatocellular carcinoma.METHODS:VX2 tumors were implanted in the liver of ten New Zealand rabbits.CT perfusion scans were made 1 week(early) and 2 weeks(late) after tumor implantation.Ten normal rabbits served as controls.CT perfusion parameters were obtained at the tumor rim,normal tissue surrounding the tumor,and control liver;the parameters were hepatic blood flow,hepatic blood volume,mean transit time,permeability of capillary vessel surface,hepatic arterial index,hepatic arterial perfusion and hepatic portal perfusion.Microvessel density and vascular endothelial growth factor were correlated.RESULTS:At the tumor rim,compared to the controls,hepatic blood flow,hepatic blood volume,permeability of capillary vessel surface,hepatic arterial index,and hepatic arterial perfusion increased,while mean transit time and hepatic portal perfusion decreased on both early and late scans(P<0.05).Hepatic arterial index increased(135%,P<0.05),combined with a sharp increase in hepatic arterial perfusion(182%,P<0.05) and a marked decrease in hepatic portal perfusion(-76%,P<0.05) at 2 weeks rather than at 1 week(P<0.05).Microvessel density and vascular endothelial growth factor showed significant linear correlations with hepatic blood flow,permeability of capillary vessel surface and hepatic arterial index,but not with hepatic blood volume or mean transit time.CONCLUSION:The CT perfusion technique demonstrated early changes of hepatic hemodynamics in this tumor model as proof-of-concept for early hepatocellular carcinoma detection in humans.

Isolated hepatic perfusion: a regional therapy for liver cancer

BACKGROUND: Many treatments have been proposed for non-resectable primary or secondary hepatic cancer but the results have been disappointing. Isolated hepatic perfusion (IHP) was attempted five decades ago but it has been ac- cepted recently after spectacular tumour responses were ob- tained by several phase trials. DATA SOURCES: An English-language literature search using MEDLINE (2003), Index Medicus (2003) and biblio- graphic reviews of books and review articles. IHP and its history and recent clinical application. RESULTS: IHP offers unique pharmacokinetic advantages for locoregional chemotherapy and biotherapy. Surgical isolation of the liver and percutaneous techniques using bal- loon occlusion catheters are reliable and safe. They appear to have significant efficacy even in patients with advanced tumor burden or those with tumors refractory to other types of therapy. CONCLUSION: IHP which has been developed in recent years is becoming a promising strategy for the treatment of unresectable liver cancer.

Quantification of angiogenesis by CT perfusion imaging in liver tumor of rabbit

BACKGROUND:Tumor angiogenesis is essential for primary and metastatic tumor growth.Computed tomography perfusion(CTP)is a new imaging method,made possible by the recent development of fast CT scanners and improved data analysis techniques,which allows measurement of the physiologic and hemodynamic properties of tissue vasculature.This study aimed to evaluate CTP in the quantification of angiogenesis and to assess the relationship between tissue perfusion parameters and microvascular density(MVD)and vascular endothelial growth factor(VEGF),attempting to detect the physiologic properties of angiogenesis.METHODS:Sixteen rabbits with VX2 liver tumors underwent multi-slice CT perfusion(MSCTP)on day 14 after tumor inoculation.CTP parameters included hepatic blood flow(HBF),hepatic blood volume(HBV),mean transit time(MTT),permeability of capillary vessel surface(PS),hepatic artery index(HAI),hepatic artery perfusion(HAP),and hepatic portal perfusion(HPP).The border of the tumor was stained with CD34 and VEGF immunohistochemical stains,and MVD was measured by anti-CD34.Then,CTP parameters were determined whether they were correlated with MVD and VEGF using Pearson’s correlation coefficient.RESULTS:The positive expression of MVD was different in the center and border of the tumor(P【0.01).There was a positive correlation between MVD and VEGF in the border(P【0.05).As more VEGF was expressed,the number of microvessels increased.Correlation analyses were also made between the perfusion parameters and MVD and VEGF in the border of the tumor.HBF,PS,HAI,and HAP values were positively correlated with MVD and VEGF(P【0.05),HPP was negatively correlated with MVD and VEGF(P【0.01),and HBV and MTT values were not correlated with MVD and VEGF(P】0.05).CONCLUSIONS:Significant correlations were found between perfusion parameters and MVD and VEGF.Therefore,MSCTP can be used to evaluate tumor angiogenesis in vivo.

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited extremely high metastatic ability including spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis,antisense approach, metalloproteinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vitro and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.

Animal models for hepatocellular carcinoma arising from alcoholic and metabolic liver diseases

Hepatocellular carcinoma (HCC) is a major and increasing cause of clinical and economic burden worldwide. Now that there are effective therapies to control or eradicate viral aetiologies, the landscape of HCC is changing with alcoholic and metabolic liver diseases becoming major catalysts. The pathogenesis of HCC is complex and incompletely understood, hampering improvements in therapy. Animal models are essential tools for advancing study on the cellular and molecular processes in HCC and for screening potential novel therapies. Many models of hepatocarcinogenesis have been established using various methods including genetic engineering, chemotoxic agents and dietary manipulation to direct implantation of tumour cells. However, none of these can accurately replicate all features found in human diseases. In this review, we provide an overview of different mouse models of HCC with a particular focus on cancer arising from alcoholic liver disease, non-alcoholic fatty liver disease and hereditary haemochromatosis. We also highlight their strengths and limitations and provide perspectives for future study.

隔离肝灌注动物实验

隔离肝灌治疗方法可以提高治疗肝癌的化疗药物局部浓度，减轻全身的毒副作用，由于手术难度大、手术操作复杂，手术技术的基本训练成为临床应用前的重要手段。为了达到熟练操作的目的，须完成大量的动物手术训练。该课题组选用大鼠作为训练材料，作为大动物手术训练前的基础训练。该文分析了大鼠手术训练死亡的原因，记录了手术中和灌注过程中大鼠的生理、病理反应，介绍了取消循环灌注泵进行手术新的方法。对５８只大鼠隔离肝灌注的手术生存率和手术并发症进行了分析。结果发现：手术操作技术、手术路径和手术器械的改进，使手术生存率大大提高，手术并发症明显减少。

兔VX2肝癌改良模型的建立及螺旋CT评价

目的探讨超声引导下经皮穿刺组织块注射法制作兔VX2肝癌模型,并与开腹组织块种植法进行比较。方法新西兰白兔24只,随机分成2组,12只/组,分别采用超声引导下经皮穿刺注射瘤组织块及开腹瘤组织块接种的方式种植于肝脏。肿瘤种植后14天,CT平扫及增强扫描与病理组织学结合评价肿瘤接种成功率及生长情况。结果肿瘤接种成功率两组均为91.7%。肿瘤最大直径穿刺组和开腹组分别为(1.04±0.12)cm和(1.09±0.14)cm,两种方法无统计学差异。肿瘤种植后发生感染率及肿瘤坏死率,开腹种植组(感染率:45.5%;坏死率:72.7%)明显高于经皮穿刺组(感染率:0;坏死率:27.3%),并有统计学意义(P<0.05)。结论超声引导下经皮穿刺注射瘤组织块建立兔VX2肝癌模型具有方法简单、成功率高、动物损伤小、肿瘤坏死率低等特点,为肝癌临床治疗和相关基础研究提供成熟的大型实验动物模型。

建立符合肝癌肝移植术后肝癌复发规律的大鼠模型

【目的】建立能反映肝癌肝移植特点的肝癌复发动物模型。【方法】选用Wistar大鼠,实验组大鼠给予肝移植术后常规联合免疫抑制方案(MP+CSA),对照组仅同法使用生理盐水,两组均接受经门静脉系统接种walker-256肿瘤细胞株手术,观察60d,死后剖腹探查取病理检查。【结果】实验组手术时间为(22.6±0.6)min,对照组为(22.1±0.7)min,两组间差异无统计学意义(P>0.05);实验组生存时间为(14.8±2.3)d,对照组为(40.9±5.3)d,两组间差异有统计学意义(P<0.05)。实验组大鼠体质量减轻量(53.5±2.6)g,较对照组体质量减轻量(34.6±5.8)g差异有统计学意义(P<0.05);实验组大鼠肿瘤复发率为95%(19/20),对照组为40%(8/20),实验组大鼠肿瘤的复发率明显高于对照组大鼠(P<0.05)。【结论】该模型能够有效模拟肝癌肝移植术后肿瘤复发的主要特征,可以作为研究肝癌肝移植术后肿瘤复发的理想大鼠模型。

CT灌注成像对兔肝肿瘤模型血管生成的定量研究

目的研究兔VX2肝移植瘤的微血管密度(MVD)和血管内皮生长因子(VEGF)与CT灌注成像各灌注参数的相关关系。方法对16只VX2肝移植瘤模型兔(种植后2周)行CT灌注扫描,获得各灌注参数值,对肿瘤周边CT强化较明显区域行VEGF、CD34的免疫组化染色,根据CD34阳性结果计数测定MVD,分析灌注参数与MVD、VEGF的相关性。结果瘤灶中心及边缘区MVD、VEGF阳性表达有明显差异(P<0.01),瘤灶边缘区MVD值随着VEGF表达增强而增高,瘤灶边缘区各灌注参数值:肝总血流灌注量(HBF)、毛细血管表面渗透性(PS)、肝动脉灌注指数(HAI)、肝动脉灌注量(HAP)与MVD、VEGF呈正相关(P<0.01),门静脉灌注量(HPP)与MVD、VEGF呈负相关(P<0.01),平均通过时间(MTT),肝血容积(HBV)与MVD、VEGF无相关性(P>0.05)。结论多层螺旋CT灌注参数与MVD、VEGF表达有明显相关性,可用于活体评价肿瘤血管生成。

稳定大鼠肝移植模型的规范及移植肝灌注方式比较

目的总结建立大鼠原位肝移植稳定模型的规范和提高成功率的措施,比较经门静脉灌注和经腹主动脉灌注对移植肝功能的影响。方法手术显微镜下,用改良的二袖套法进行100例大鼠原位肝移植,并依据灌注方式分组:经门静脉灌注和经腹主动脉灌注组(n=50)。检测术后肝功能,观察组织病理学改变、术后生存率以及手术并发症。结果两组组织病理学表现相同。手术成功率分别为98%(49/50)和96%(48/50),3月存活率分别为93.5%(29/31)和93.3%(28/30),术后肝功能、手术成功率和3月存活率差异无显著性(P>0.05)。结论供肝经门静脉灌注和经腹主动脉灌注均可采用,应根据研究目的选择适当肝移植模型。熟练的显微外科技术、规范细致的手术操作、缩短无肝期时间是提高成功率、减少术后并发症的关键。

CT灌注成像及MR扩散加权成像评价兔肝纤维化的价值

目的探讨双源CT灌注成像及MR扩散加权成像评价兔肝纤维化分期的价值。方法新西兰大白兔60只进行四氯化碳腹腔注射建立肝纤维化模型(实验组),新西兰大白兔10只进行腹腔注射生理盐水建立对照模型(对照组)。行双源CT灌注成像及MR扩散加权成像,比较不同肝纤维化分期时CT灌注参数,包括门静脉灌注量(portal venous perfusion,PVP)、肝动脉灌注指数(hepatic perfusion index,HPI)和MRI扩散加权成像测量ADCperf值的变化,分析各参数与纤维化分期的关系。结果 HPI随着肝纤维化程度的增加逐渐升高,PVP随着肝纤维化程度的增加而减低。PVP与肝纤维化的严重程度呈负相关(r=-0.589),HPI与肝纤维化的严重程度呈正相关(r=0.652)。ROC曲线显示,PVP预测S2期及以上肝纤维化时诊断效能最佳,HPI预测S2期及以上肝纤维化时诊断效能最佳。随着肝纤维化程度加重,ADCperf值依次降低,差异有统计学意义(P<0.01),ADCperf预测S2期及以上肝纤维化时曲线下面积最大。肝脏ADCperf值与肝纤维化的严重程度呈负相关(r=-0.720)。结论HPI、PVP、ADCperf能够反映肝纤维化各期灌注变化。

大鼠肝纤维化和肝硬化MR-PWI参数与CD34和a-SMA的相关性

目的：探讨大鼠肝纤维化、肝硬化模型磁共振灌注成像（MR-PWI）参数的意义及其与CD34、a-SMA的相关性。方法：清洁级SD大鼠随机分为实验组（n=84）和对照组（n=16）。实验组采用硫代乙酰胺（TAA）腹腔定点注射,每次200mg/kg,3次/周,对照组同期腹腔注射同剂量的生理盐水。建模第6~30周后实验租和对照组分批次行肝脏磁共振常规扫描及灌注成像。病理上将鼠肝损伤分为肝纤维化0期、Ⅰ~Ⅱ期和Ⅲ~Ⅳ期、肝硬化结节期,并与灌注参数TTP、WIR、WOR和免疫组化指标CD34、a-SMA表达作对照。结果：共有69只大鼠（实验组57只,对照组14只）成功获取MR-PWI参数,成功率69%（69/100）。实验组门静脉和肝实质的TTP随着肝纤维化及肝硬化进展而延长,而门静脉WIR、WOR在肝纤维化Ⅰ~Ⅱ期和Ⅲ~Ⅳ期下降不明显,肝硬化结节期则明显下降;肝实质WIR、WOR则随着肝纤维化、肝硬化进展而逐渐下降,各期之间差异有统计学差异（P〈0.05）;各灌注参数与肝纤维化病理分期之间有较好相关性（P〈0.05）。CD34和a-SMA表达在肝纤维化Ⅰ~Ⅱ期不明显,肝纤维化Ⅲ~Ⅳ期和肝硬化结节期则明显表达,与对照组比较,实验组肝纤维化Ⅲ~Ⅳ期和肝硬化期CD34、SMA表达均显著增加（P〈0.01）。门静脉及肝实质灌注参数TTP、WIR、WOR和CD34、a-SMA表达有相关性（P〈0.05）。结论：门静脉和肝实质TTP、WIR、WOR与大鼠肝纤维化、肝硬化不同分期之间有相关性,与CD34、a-SMA表达有相关性。

实验性小鼠前胃癌肝转移模型的建立

目的 :建立实验性小鼠前胃癌肝转移模型。方法 :将浓度为 1× 10 7个·ml-1的小鼠前胃鳞癌细胞 (MFC)悬液 0 2ml接种于 615小鼠脾被膜下 ,分别于术后第 1、2、3、4周处死 2～ 3只小鼠 ,观察腹腔内肿瘤生长情况 ,其余小鼠待其自然死亡 ,观察生存天数。结果 :肝转移发生率为 83 3 3 % ,小鼠平均荷瘤生存时间为 (3 7 83± 7 3 2 )d。病理结果提示肝转移瘤细胞与脾脏肿瘤细胞形态学相似 ,符合低分化鳞癌的特征。结论 :本研究成功建立了实验性小鼠前胃癌肝转移模型 。

大鼠肝癌细胞C2不同途径接种造模及比较

目的探讨大鼠肝癌细胞系C2肝、脾原位接种成瘤及肠系膜静脉注射在不同内环境情况下对肿瘤发生和侵袭转移能力的影响,并筛选最佳C2肝细胞癌肺转移动物模型。方法 C2进行体外培养。将雌性BALB/cA裸鼠随机分为肝、脾原位接种及肠系膜静脉注射3组。观察裸鼠肝、脾成瘤率、裸鼠存活率、肿瘤病理学特点及远处脏器转移率及强度。结果肝原位接种组成瘤率为100%,肿瘤呈结节状膨胀性生长,周围肝组织和腹壁有粘连和侵袭,肺转移率为100%。脾原位接种组脾成瘤率为50%,肉眼未见明显的肿瘤形成,但部分肝表面可见转移灶;镜下肝、肺肿瘤转移率分别为67%和33%。肠系膜静脉注射组肉眼未见转移灶,镜下示肝、肺肿瘤转移率均为25%。结论三种不同方法构建的C2肝癌肺转移动物模型均有肺转移灶的出现,其中,肝原位接种模型最适合于肝细胞癌体内及肺转移机制及药物干预研究。

耐药VX2肝癌模型的建立

目的 探讨应用经化疗药物诱导后传代的VX2肿瘤建立移植性耐药肝癌模型的可行性。方法 将 2 0只大耳白兔随机分为 4组 ,建立肝癌模型。A、B组和C、D组植入肿瘤分别取自常规传代和药物诱导后的传代兔。模型建立 3周后 ,增强CT检查确定肿瘤大小。直视下穿刺肝动脉 ,A、C两组注入阿霉素 (4mg/2ml) ,B、D两组注入等量生理盐水。 1周后CT复查 ,比较各组肿瘤倍增率。流式细胞仪检测各组肿瘤凋亡率。结果 模型建立 3周后各组肿瘤大小无明显差异。治疗 1周后增强CT复查 ,各组肿瘤体积均增大。其中 ,A组肿瘤倍增率显著低于其他组 (P <0 .0 5 ) ;C组低于B、D组 ,但各组间差异无显著性 (P >0 .0 5 )。A组肿瘤凋亡率显著高于其他组 (P <0 .0 5 ) ;C组高于B、D组 ,但组间差异无显著性 (P >0 .0 5 )。结论 通过药物诱导建立的移植性兔VX2肝癌模型具有明显耐药特征 ,可作为耐药肝癌模型用于肝癌全身与介入性药物治疗的实验研究。

兔VX2肝癌HIFU治疗前后肿瘤新生血管MSCT灌注成像研究

目的通过对兔肝VX2种植瘤高强度聚焦超声(HIFU)治疗前后MSCT灌注参数与肿瘤微血管密度(MVD)、血管内皮生长因子(VEGF)表达的相关性研究,探讨MSCT灌注成像活体评价肝肿瘤血管生成的价值。方法 30只VX2肝种植瘤模型兔,分为对照组和治疗组,治疗组进行HIFU治疗,分别对两组试验动物行MSCT灌注成像检查,并用免疫组织化学方法测定MVD、VEGF表达情况,分析MSCT灌注参数与MVD和VEGF的相关性。结果治疗组灌注参数、MVD及VEGF表达程度均明显低于对照组,VEGF和MVD的表达呈显著正相关,MSCT灌注参数中,肝总血流灌注量(HBF),毛细血管表面通透性(PS)和肝灌注指数(HAI)与VEGF、MVD的表达均呈高度正相关,而肝血容积(HBV)、平均通过时间(MTT)与VEGF、MVD的表达无相关性。结论 MSCT灌注参数与MVD、VEGF表达有明显相关性,可用于活体评价肿瘤血管生成。

猪肝脏体外不同灌流系统建立和评价

目的:探讨体外肝脏灌流系统中的影响因素,建立稳定有效的体外肝脏灌流系统。方法:根据灌流和氧合的方式建立不同的体外猪肝脏灌流系统,随机分为3组(每组n=4)。A组单独灌流门静脉,氧合灌流液;B组同时灌流门静脉和肝动脉,共同氧合门静脉和肝动脉的灌流液;C组同时灌流门静脉和肝动脉,分别氧合门静脉和肝动脉的灌流液。观察体外肝脏灌流时间、胆汁分泌量、病理变化和血液动力学等指标。结果:A组的灌流时间明显低于B组和C组。在1,3,6h的时间点A组胆汁分泌量、血液动力学等指标和B、C组比较差异有统计学意义。在12h时间点B组和C组胆汁分泌量和血液动力学的差异也有统计学意义。结论:同时灌流门静脉和肝动脉,分别氧合门静脉和肝动脉灌流液的灌流系统更稳定,对体外肝脏功能的维持效果较好。

二乙基亚硝胺诱发大鼠肝细胞癌发生过程中基质金属蛋白酶动态变化

目的研究基质金属蛋白酶(MMPs)在肝细胞癌形成过程中动态变化及作用方法采用二乙基亚硝胺(DENA)诱癌建立大鼠肝癌模型.应用明胶酶谱法测定诱癌过程中癌变发生的三个阶段6个时相肝组织中MMP-2和MMP-9的表达及活性.结果肝癌发生过程中基质金属蛋白酶呈持续升高的趋势,至早期癌变期及肿瘤演进期(16wk～24wk)MMP-2,MMP-9的表达呈一个较高的平台(分别为167350±20436和237754±32675),且发现MMP-2的活性形式表达显著增加.结论 MMPs的表达不同与肝星状细胞的激活和肝细胞癌形成有关,而与肿瘤的结节数及大小无关.