腹腔移植微载体粘附人肝细胞对急性肝衰竭小鼠的保护作用

为了探讨腹腔移植微载体粘附培养人肝细胞对D氨基半乳糖(GalN)诱导的急性肝衰竭小鼠的保护作用．本实验采用胶原酶灌流法分离人肝细胞，胶原被覆的微载体Cytodex3加以培养，经腹腔注射肝细胞及载体，观察其对急性肝衰竭小鼠的影响。结果显示．与只接受微载体而无粘附肝细胞的对照组比较，腹腔移植培养肝细胞对肝衰竭小鼠的存活率有明显改善，其血清ALT及总胆红素较低(P<0．05)。肝组织病理改变较轻。该初步观察结果提示．腹腔移植微载体牯附培养肝细胞可提供代谢支持作用，从而使GalN损伤的小鼠肝功能得以恢复。     

牛肝细胞刺激物质的成分及对小鼠急性肝衰竭的保护作用

应用液相色谱及等离子体发射光谱法分析新生牛肝匀浆提取液中的游离氨基酸及微量元素成分;观察该提取液腹腔注射后对CCl_4中毒小鼠急性肝衰竭的保护作用。结果表明提取液中含有13种以上的游离氨基酸,其中6种为必需氨基酸,牛磺酸含量为正常成人血浆的8倍,支/芳氨基酸比值为3.2。微量元素以Mn、Zn、Mg、Se及Fe的含量较高。该提取液能明显提高实验动物的成活率(60％,对照组为10％,P相似文献   

肝细胞移植治疗急性肝衰竭

“20世纪是药物治疗的年代，21世纪却是细胞治疗的年代”。细胞治疗是指将干细胞或由其分化产生的功能细胞植入病变部位代偿病变细胞丧失的功能，或将细胞经体外遗传操作后用于疾病治疗的方法。急性肝衰竭（acute liver failure，ALF）常规药物治疗疗效差，是一种病死率极高的临床综合征，在我国主要由病毒性肝炎引起。近几年开展的较为有效的治疗方法主要有原位肝移植、人工肝支持系统和肝细胞移植（hepatocyte transplantation，HTx）。肝移植被认为是目前治疗ALF的最有效方法，但供肝短缺、免疫排斥和高额费用限制了受益者范围。     

抗TNFR1中和抗体对急性肝衰竭小鼠肝细胞的保护作用

目的探讨抗TNFR1中和抗体对急性肝衰竭小鼠肝细胞的作用。方法联合应用D-氨基半乳糖和肿瘤坏死因子-α造模，酶联免疫吸附试验测定血清ALT、AST浓度。HE染色检查肝脏组织病理变化、脱氧核糖核苷酸转移酶介导的缺口末端标记（TUNEL）技术检测肝细胞的凋亡程度。结果模型组各时间点（4、8、12h）均可见明显的肝细胞凋亡，治疗组各时间点亦可见少量凋亡细胞，但与模型组比较各时间点均明显减少（P〈0．01），其血清ALT、AST浓度较模型组也有下降。结论抗TNFR1中和抗体对TNF—α能诱导肝细胞损伤具有保护作用肝。     

急性肝衰竭小鼠肝细胞核脱氧核糖核酸损伤的研究

目的:研究急性肝衰竭小鼠肝细胞核DNA(脱氧核糖核酸)的损伤变化及影响因素.方法:健康雌性昆明小鼠腹腔注射D-氨基半乳糖(D-Galn)和脂多糖(LPS)制作急性肝衰竭模型,采用慧星实验检测注药后0.5小时、1小时、2小时、4小时、6小时和8小时小鼠肝细胞核DNA的损伤,ELISA检测血清肿瘤坏死因子-α(TNF-α)浓度.结果:①模型组小鼠于注药0.5小时Olive尾矩值即明显增大,且随时间延伸逐渐增加,与正常组比较,差异均具有显著性意义(P<0.05).②模型组小鼠于注药0.5小时血清TNF-α浓度无升高;1小时略有升高,但与正常对照组比较,差异无显著性意义;2小时以后,升高趋势明显,各组与正常对照组比较,差异均具有显著性意义.结论:急性肝衰竭小鼠肝细胞核DNA较早出现损伤,并随时间延伸逐渐加重,最终可能诱发了肝细胞凋亡;血清TNF-α浓度的升高与肝细胞核DNA的损伤不完全同步,但其可能具有促进肝细胞DNA损伤的作用.     

维生素D对急性肝衰竭小鼠肝脏的保护作用

目的:通过小鼠动物模型探讨维生素D对急性肝衰竭肝脏的保护作用。方法:应用D-氨基半乳糖（D-GalN）联合脂多糖（LPS）诱导的小鼠急性肝衰竭模型,观察长期维生素D缺乏对肝脏损伤、肝脏炎症信号的影响。采用硫代乙酰胺（TAA）诱导的小鼠急性肝衰竭模型,观察维生素D缺乏对模型小鼠存活率的影响,并观测补充高剂量维生素D对模型...     

移植培养人肝细胞对急性肝衰竭小鼠的保护作用

目的探讨腹腔移植微载体粘附培养人肝细胞对Ｄ氨基半乳糖（ＤＧａｌ）诱导的急性肝衰竭小鼠的保护作用．方法采用胶原酶灌流法分离人肝细胞，胶原被覆的微载体Ｃｙｔｏｄｅｘ３加以培养，经腹腔注射２×１０６个肝细胞及载体，观察急性肝衰竭小鼠（ｎ＝３２，２５ｇ／ｋｇＤＧａｌ诱导４ｈ）７２ｈ存活率、血清ＡＬＴ、总胆红素以及肝脏病理变化．结果与只接受微载体而无粘附肝细胞的对照组比较，腹腔移植培养肝细胞对肝衰竭小鼠的存活率有明显改善（６５％ｖｓ０％），其血清ＡＬＴ（ＩＵ／Ｌ，２１７４４±２６３０ｖｓ４２６３１±４９２８，Ｐ＜００５）及总胆红素（ｍｍｏｌ／Ｌ，２６９１±３７６ｖｓ４１６８±３８３，Ｐ＜００５）较低．肝组织病理改变较轻．结论腹腔移植微载体粘附培养肝细胞可提供代谢支持，使ＤＧａｌ损伤的小鼠肝功能恢复．     

肝细胞移植治疗肝衰竭研究进展

约有17.5%的肝衰竭患者需行肝移植治疗。虽然肝移植已经广泛开展,并且术后一年生存率可达80%,但限于供体肝脏来源困难和昂贵的治疗费用,难以普及使用。肝细胞移植则是治疗暴发性肝衰竭与遗传性肝病的另一治疗手段,已经有数名患者为恢复肝功能而接受了这种细胞移植。肝细胞移植与原位肝移植相比具有以下几方面优势:1.同一来源的肝细胞可以用于多个受体;2.不需要手术,也没有整肝移植的高死亡率;3.可以临时应用起过渡作用,以等待肝脏复原或移植。     

肝衰竭患者血浆(血清)对体外培养肝细胞毒性作用的研究现状

生物人工肝是以培养肝细胞为基础的人工肝支持系统,高活性和功能良好的肝细胞是其基本要素之一,然而,由于肝衰竭患者肝脏大量肝细胞坏死,导致机体代谢紊乱和大量毒性物质堆集,这些毒性物质会对反应器中肝细胞产生毒性作用,直接影响生物人工肝的治疗效果.因此,了解肝衰竭患者血浆（liver failure plasma,LFP）或血清（liver failure serum,LFS）对体外培养肝细胞的各种不良作用,对生物人工肝的临床应用有重要意义.本文综述近年来在该领域的研究进展.     

培养胎肝细胞上清对暴发性肝衰竭小鼠存活率的影响

目的探讨胎肝细胞悬液治疗严重肝病的机理以及使用培养胎肝细胞分泌上清（ＦＨＣＳ）替代的可能性．方法采用Ｄ＿氨基半乳糖（Ｄ＿ｇａｌｎ）／内毒素（ＬＰＳ）诱发小鼠暴发性肝衰竭（ＦＨＦ），观察ＦＨＣＳ的保护作用．结果ＦＨＣＳ能显著提高ＦＨＦ小鼠的存活率（Ｐ＜００１），降低血清转氨酶（Ｐ＜００５），减轻肝细胞坏死程度，并促进肝细胞再生．结论ＦＨＣＳ对ＦＨＦ有较好的保护与治疗作用，可望为已被迫放弃的胎肝细胞悬液输注疗法开辟新的途径     

Liver transplantation for acute hepatic failure

There are numerous causes of acute hepatic failure (AHF). Cerebral edema, coagulopathy, renal failure, metabolic disturbances and infection are the main clinical sequelae. Patients with AHF should be stabilized when first encountered and transferred to the nearest liver transplant center, as AHF progresses quickly and is often fatal. There are few adequate medical interventions and care of patients with AHF is supportive until spontaneous recovery ensues. If recovery does not appear to occur, most causes of AHF are well accepted indications for liver transplantation.     

An experimentally-induced acute hepatic failure model in various strains of mice

When BALB/cAJcl mice are intravenously injected with heat-killedPropionibacterium acnes (P. acnes) followed by an intravenous injection of lipopolysaccharide (LPS) 7 days later, massive necrosis is induced in the liver tissue and most of the mice die within 24 hours of LPS injection. Using this experimental model, acute hepatic failure was induced in various strains of mice and the difference in the response was studied. As a result, as in BALB/cAJcl mice, acut hepatic failure was also induced in BALB/ cAJcl-nu, AKR/J, C3H/HeNJcl, C57BL/6NJcl and DDy mice. However, as an exception, hepatic cell necrosis was hardly seen and the survival rate was remarkable high in C3H/HeJ mice, which genetically do not respond to LPS stimulation. These results indicate that for this experimental induction of acute hepatic failure, macrophages must be activated by the two-step stimulation ofP. acnes and LPS.     

Orionin对急性肝衰竭小鼠的保护作用及其对肝组织细胞因子水平的影响*

目的 探讨冬凌草甲素（Oridonin）对脂多糖/D-氨基半乳糖氨（LPS/D-Gal）联合诱导的急性肝衰竭（ALF）小鼠的保护作用及其对肝组织细胞因子水平的影响。方法 取150只小鼠,随机分成5组,每组 30 只。采用LPS/D-Gal腹腔注射建立小鼠ALF模型,设生理盐水对照组、Oridonin对照组、LPS/D-Gal诱导模型组和LPS/D-Gal处理及不同剂量Oridonin干预组。采用Real-time PCR法检测肝组织肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1α、IL-1β和IL-6 mRNA水平。结果 模型组小鼠48 h存活率为0.0% （0/30）,而两个Oridonin干预组小鼠48 h存活率分别提高至64.5% （19/30）和80.6% （24/30,P<0.01）; 组织病理学检查显示模型组小鼠肝细胞呈大块或/和亚大块坏死,肝小叶结构消失,残存肝细胞肿胀、空泡变性,肝窦肿胀充血,炎细胞浸润。Oridonin干预组小鼠肝细胞坏死、空泡变性和炎细胞浸润等较模型组有明显的改善;模型组小鼠血清ALT和AST水平分别为（345.3 ± 54.1） U/L和（500.2±53.5） U/L,明显高于对照组的[（42.3±0.6）U/L和（117.1±9.8）U/L,P<0.01],两个Oridonin干预组分别为 （303.9±39.5） U/L和（340.6±2.8） U/L及[（130.2±38.3） U/L和 （209.8±36.2） U/L,P<0.05];模型组小鼠肝组织TNF-α、IL-1α、IL-1β和IL-6 mRNA水平显著高于正常对照组（P<0.01）,而两个Oridonin干预组肝组织TNF-α、IL-1α、IL-1β和IL-6 mRNA水平显著低于模型组 （P<0.01）。结论 Oridonin对LPS/D-Gal诱导的ALF小鼠具有显著的保护作用,其作用机制可能与降低肝组织细胞因子水平有关。     

A double-blinded,randomized trial of hydrocortisone in acute hepatic failure

The Acute Hepatic Failure Study Group (AHFSG) has conducted a double-blinded, randomized evaluation of hydrocortisone in patients with acute hepatic failure. From July 1975 through August 1978, a 38-month period, 18 medical centers in the United States and one in Canada participated in this trial. A total of 64 patients were accessed and found eligible to participate in the study; two of them were subsequently eliminated from our analysis. Eighteen patients received placebo; 23 received 400 mg hydrocortisone per day, and 21 patients were administered 800 mg hydrocortisone per day. We did not observe any therapeutic effect of hydrocortisone, and the survival rates for placebo versus 400 mg and versus 800 mg hydrocortisone per day were 22%, 9%, and 24%, respectively. Fulminant hepatitis associated with drug hepatotoxicity or non-A, non-B hepatitis seemed to have a worse prognosis than fulminant B, although these differences were not significant. Serum -fetoprotein had a modest prognostic value of survival and seemed to be limited to fulminant B. The AHFSG recommends, therefore, that corticosteroid use in acute hepatic failure with hepatic encephalopathy be discontinued.Supported in part by grants from the National Institute of Health (HL-23420 and CA-30209)     

Effects of irsoglandine maleate in an experimentally-induced acute hepatic failure model using mice

When heat-killedPropionibacterium acnes was intravenously injected into mice followed by an intravenous injection of a small amount of Gram-negative lipopolysaccharide seven days later, most of the mice died of massive hepatic cell necrosis within 24 hours. However, when irsoglandine maleate, an antiulcer agent, was administered to mice during the period of experimental induction of acute hepatic failure, the survival rate, serum transaminase levels and histological changes of the liver remarkably improved. These results suggested that irsoglandine maleate may have protective effects on the liver in our experimentally-induced acute hepatic failure model using mice. Therefore, in the absence of a definitive therapy for fulminant hepatitis, irsoglandine maleate may be a promising therapeutic agent.     

Insulin and glucagon therapy of acute hepatic failure

When insulin and glucagon are administered to rats with severe liver injury, survival is enhanced with an attenuation of the liver injury compared to that of untreated controls. In rats with acute liver injury both hormones produce a rapid normalization of hepatic protein content following initiation of DNA synthesis. When rats receive both hormones after partial hepatectomy, the first burst of DNA synthesis reaches a maximum earlier than that seen in controls. Both hormones enhance the increment of hepatic putrescine essential for DNA synthesis through activation of ornithine decaroxylase and/or spermidine-N¹-acetyltransferase. The enhancement of putrescine content by each hormone is additive. Putrescine supplementation promotes hepatic DNA synthesis after hepatectomy. Based on these data, we conclude that a combination of insulin, and glucagon is effective in the therapy of acute hepatic failure in rats. The restoration of liver function as well as the stimulation of liver cell proliferation via putrescine production may contribute to this effect.Presented at the Proceedings of the International Meeting on Normal and Neoplastic Growth in Hepatology, Bari, Italy, June 1989.