龟板促骨髓间充质干细胞肝脏归巢的作用研究

目的 探讨龟板饲养对移植骨髓间充质干细胞(MSCs)肝脏归巢的作用.方法 密度梯度离心法分离、培养大鼠MSCs,带绿色荧光蛋白腺病毒转染标记移植MSCs,激光共聚焦检测肝脏组织MSCs分布,Western blot检测肝脏肝细胞生长因子(HGF)蛋白及MSCs c-met蛋白表达,携带siRNA腺病毒转染下调c-met表达.结果 龟板饲养较普通饮食明显增加移植MSCs肝脏归巢(P＜0.05);同时,龟板饲养增加大鼠肝脏HGF蛋白表达,阻断HGF/c-met可部分逆转龟板促MSCs归巢的作用.结论 龟板饲养可能通过HGF/c-met轴促MSCs向肝脏组织归巢.     

甘草酸二铵对肝纤维化大鼠肝脏脂质过氧化与间质性胶原酶活性的影响

目的：探讨甘草酸二铵影响纤维化肝脏脂质过氧化与间质性胶原酶活性的抗肝纤维化作用机制。 方法： 以四氯化碳(CCl4)皮下注射与高脂肪低蛋白饮食复合因素诱导大鼠肝纤维化模型，而后予以甘草酸二铵口服治疗，正常大鼠与模型对照组给予等量生理盐水。HE染色与胶原染色观察大鼠肝组织炎性坏死与胶原沉积病理改变，按试剂盒方法检测血清肝功能（ALT、AST、Alb与总胆红素等）变化，检测肝组织主要过氧化损伤指标：超氧化物歧化酶（SOD）活性, 丙二醛（MDA）含量，谷胱甘肽（GSH）含量与谷胱甘肽过氧化物酶（GSH-Px）活性，水解法测定肝组织羟脯氨酸含量，酶底物反应法分析肝组织间质性胶原酶活性变化，RT-PCR法分析肝组织Ⅰ型前胶原基因表达。 结果： 与正常大鼠相比，模型大鼠肝脏有明显胶原沉积与肝纤维化，伴有不同程度的肝细胞炎性损伤坏死；甘草酸二铵药物组明显减轻模型大鼠肝组织损伤坏死与胶原沉积等病理变化。模型大鼠肝功能指标,包括血清总胆红素含量、ALT与AST活性、白蛋白含量均明显减少（P<0.01）。药物组大鼠血清总胆红素含量、AST与ALT活性显著低于模型组（P<0.05），而白蛋白含量高于模型组（P<0.05）。此外，药物组大鼠肝组织羟脯氨酸含量(151.3±37.3 μg/g)明显少于模型组(170.9±15.3 μg/g，P<0.05)，Ⅰ型前胶原基因表达弱于模型组（P<0.05）；肝组织MDA含量(1.96±0.23 μmol/g)低于模型组(2.44±0.32 μmol/L, P<0.05)，SOD水平(19.60±0.97 NU/g)高于模型组(20.60±0.33 NU/g，P<0.05)，GSH含量(47.0±9.1 g/g)与GSH-Px活性(53.1±4.1 U/g)高于模型组(41.2±3.5 g/g；46.7±6.1 U/g，P<0.05)；肝组织间质性胶原酶活性(43.89±7.74 U)高于模型组(32.01±2.75 U，P<0.05)。 结论： 甘草酸二铵有明显抗肝纤维化大鼠肝脏脂质过氧化损伤与提高肝组织间质性胶原酶活性的作用，该作用是药物抗肝纤维化的重要机制。     

金丝桃苷对CCl4诱导大鼠急性肝损伤抗氧化应激研究

目的研究金丝桃苷（Hyp）对四氯化碳（CCI。）诱导大鼠急性肝损伤的治疗作用。方法采用大鼠CCl4急性肝损伤模型,观察Hyp对急性肝损伤大鼠肝脏组织病理学改变的影响;检测肝组织匀浆中超氧化物歧化酶（T-SOD）、谷胱甘肽（GSH）的活性及丙二醛（MDA）含量变化。结果CCl4模型组大鼠肝组织HE染色病理检测结果见明显炎症变性死及纤维组织增生现象;Hyp高剂量60mg／kg、中剂量30mg／kg治疗组的肝组织病理改变明显改善;Hyp治疗组肝组织中T—SOD、GSH活性明显升高,MDA含量明显降低,并存在量效关系。结论Hyp对CCl4引起的大鼠急性肝损伤有较好的治疗作用,其机制可能与其抗氧化活性有关。     

肝炎康对D-氨基半乳糖所致大鼠急性肝损伤的保护作用

目的:观察肝炎康对D-氨基半乳糖(D-GalN)所致大鼠急性肝损伤的保护作用。方法:建立D-GalN致大鼠急性肝损伤模型,设正常对照组,模型对照组,云芝多糖阳性对照组和肝炎康大、中、小剂量治疗组。检测大鼠血清中谷丙转氨酶(ALT)、天门冬氨酸转氨酶(AST)活性和肝组织均浆中丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性,并观察肝脏组织病理学变化,以评价肝炎康对肝脏的保护作用。结果:肝炎康能明显降低由D-GalN所致急性肝损伤大鼠血清中ALT、AST活性,降低肝组织中MDA含量,提高SOD活性,并能明显改善肝脏组织病理损伤。结论:肝炎康对D-GalN所致大鼠急性肝损伤具有一定的保护作用。     

重组人肝再生增强因子可逆转免疫损伤性肝纤维化

目的:了解重组人肝再生增强因子(hALR)抗免疫损伤性肝纤维化的活性。方法:建立人血白蛋白免疫损伤性大鼠肝纤维化模型。模型完成后予hALR 腹腔注射。观察肝纤维化大鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)水平,肝组织胶原蛋白含量及肝脏病理学改变。结果:重组人肝再生增强因子(hALR)可降低肝纤维化大鼠的ALT、AST、LDH水平;肝组织胶原蛋白含量的测定表明hALR治疗组大鼠肝组织胶原含量明显低于模型组及阴性对照组;病理组织切片也发现hALR治疗组大鼠肝纤维化程度较模型组及阴性对照组明显减轻。结论:重组人肝再生增强因子(hALR)可逆转免疫损伤性大鼠肝纤维化。     

熊果酸对肝纤维化大鼠NOX2/ROS/NLRP3炎性小体活化的影响

目的观察熊果酸(UA)对肝纤维化模型大鼠肝内胶原沉积的改善作用及其对NADPH氧化酶2(NOX2)/活性氧簇(ROS)/NLRP3炎性小体活化的影响。方法将大鼠随机分为对照组、CCl4模型组及UA治疗组。用CCl4诱导构建SD大鼠肝纤维化模型,一半用作UA治疗组。用试剂盒检测血清ALT含量;HE染色观察肝脏病理;天狼星红染色观察肝内胶原沉积;RT-q PCR法检测Nox2、Nlrp3、caspase1及IL-1βmRNA表达量;Western blot及免疫组化检测肝脏中NOX2、NLRP3、casepase-1 p45、caspase-1 p10及IL-1β蛋白表达;DCFH-DA荧光探针法检测肝脏组织内ROS水平。结果与对照组相比,CCl4模型组血清ALT水平升高(P0.05),Ishak's肝纤维化评分明显上升,胶原沉积明显增多(P0.05),Nox2、Nlrp3、Caspase1及IL-1βmRNA的表达水平明显上升(P0.05);NOX2、NLRP3、caspase-1 p10及IL-1β蛋白的表达均出现明显增加,肝组织内ROS含量增加(P0.05);与CCl4模型组相比,UA治疗组血清ALT含量下降(P0.05),肝脏Ishak's肝纤维化评分及胶原沉积减少(P0.05),Nox2、Nlrp3、caspase1及IL-1βmRNA的表达水平明显下降(P0.05),NOX2、NLRP3、caspase-1 p10及IL-1β蛋白表达明显下降,肝脏组织内ROS水平显著改善(P0.05)。结论 UA减轻肝脏炎性反应并减少肝内胶原沉积,其机制可能与其抑制肝纤维化大鼠肝内NOX2/ROS/NLRP3炎性小体活化及IL-1β的释放减少有关。     

益气化瘀柔肝方通过调控NLRP3炎性小体活性缓解CCl_4诱导的大鼠肝纤维化

目的研究益气化瘀柔肝方(Yiqi Huayu Rougan Recipe, YHR)对CCl_4诱导的大鼠肝纤维化的影响及分子机制。方法将SD大鼠随机分为正常组,模型组,秋水仙碱组,YHR低、中、高剂量(5.4、10.8、21.6 g/kg)组。末次给药后,收集大鼠血清和新鲜肝脏组织,运用生化法检测大鼠血清ALT、AST、ALP含量;ELISA法检测血清HA、LN、Hyp和促炎性因子(TNF-α、IL-6、IL-1β)水平;采用HE染色和Masson染色法观察大鼠肝组织病理变化;采用RT-qPCR和Western blot法检测肝组织TLR4、NF-κB和炎性小体相关分子的mRNA和蛋白表达水平。结果秋水仙碱和各剂量组的YHR均能不同程度的减少大鼠血清酶谱含量以及肝纤维化指标水平,降低促炎性因子水平,且YHR高剂量组最为明显;秋水仙碱组和YHR各剂量组均能显著改善肝纤维化大鼠的病变程度,且肝组织中的TLR4、NF-κB和炎性小体相关分子的表达均减少。结论 YHR具有缓解CCl_4诱导的大鼠肝纤维化的作用,其药效作用机制可能与调控NLRP3炎性小体信号通路活性有关。     

肝纤维化大鼠肝脏中组蛋白修饰水平的变化

目的:观察肝纤维化大鼠肝脏中组蛋白修饰的变化,并探讨其在肝纤维化发生发展过程中可能的作用。方法:雄性Wistar大鼠20只,随机分为正常对照组和肝纤维化组,其中肝纤维化组采用CCl_4皮下注射以制备大鼠肝纤维化模型,正常组注射等量植物油溶液。实验第8周末,股动脉放血处死大鼠,取2组血清,采用生化和放射免疫法测定血清肝功能指标丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST),以及肝纤维化标志物血清透明质酸(HA)、层粘连蛋白(LN)、Ⅳ型胶原(Col Ⅳ)和Ⅲ型前胶原(PCⅢ)的水平;取2组大鼠肝脏,测定肝脏指数;取肝组织常规固定,HE染色和Masson染色观察组织病理改变及胶原纤维沉积情况;Western blot检测2组大鼠肝脏组织中α-平滑肌肌动蛋白(α-SMA)和I型胶原(ColⅠ)表达情况,以及acH4K12、acH3K9、H3K4me2和H3K9me2修饰水平的变化。结果:与对照组相比,模型组大鼠肝脏指数及ALT、AST、HA、LN、ColⅣ和PCⅢ水平明显增高(P0.05);Western blot检测发现,与对照组比较,肝纤维化组大鼠肝组织的acH4K12修饰水平减少(P0.05),acH3K9和H3K9me2修饰水平及α-SMA和ColⅠ表达明显增加(P0.05),H3K4me2修饰水平的差异无统计学显著性。结论:肝纤维化大鼠肝脏中acH4K12、acH3K9和H3K9me2修饰水平改变可能与某些细胞外基质代谢相关基因转录调控有关,从而参与了大鼠肝纤维化发生。     

龟板含药血清促进骨髓间充质干细胞骨形态发生蛋白4的表达

目的 龟板有促大鼠骨髓间充质干细胞(MSCs)增殖作用,本研究观察龟板含药血清对MSCs中骨形态发生蛋白4(BMP4)表达的影响.方法 使用密度梯度法分离大鼠骨髓间充质干细胞进行培养,传代纯化,在基础培养液中添加不同浓度龟板含药血清和对照血清,用荧光定量PCR和原位杂交方法检测BMP4 mRNA表达,用ELISA、免疫组织化学和免疫荧光组织化学法结合激光扫描共聚焦显微镜技术检测BMP4蛋白表达的变化.结果 BMP4蛋白和mRNA表达相一致,龟板含药血清以浓度依赖方式促进MSC的BMP4 mRNA及其蛋白表达.结论 龟板含药血清以时效和量效依赖性促进BMP4在MSCs的表达,可能与龟板促MSCs增殖作用有关.     

茶多酚对酒精诱导的小鼠肝脂质过氧化和血清ALT活性变化的影响

目的: 研究茶多酚(TP)对酒精性肝损伤的防治作用。方法: 通过离体和整体实验,采用分光光度法和血清生化检测法,检测肝组织脂质过氧化产物丙二醛(MDA)水平和血清谷丙转氨酶(ALT)的活性。结果: 离体实验中,TP+酒精组肝MDA水平明显低于生理盐水+酒精组(P＜0.01)。整体实验中,TP+酒精组小鼠肝脏MDA水平和血清中ALT活性均显著低于生理盐水+酒精组(P＜0.05)。此外,灌酒精1h后再给TP组,小鼠肝脏MDA含量也明显低于酒精+生理盐水组(P＜0.01)。结论: 茶多酚能抑制酒精引起的小鼠肝组织MDA水平和血清ALT活性的升高,可能具有防治酒精性肝损伤的作用。     

Role of liver biopsy in autoimmune liver disease

This article will review histological aspects of three chronic liver diseases – autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) – in which autoimmune mechanisms are thought to be involved. The changing role of liver biopsy in the diagnosis and management of patients with autoimmune liver disease will also be discussed. In the case of autoimmune hepatitis, histological assessments remain important in establishing a diagnosis, identifying prognostic features and monitoring therapeutic responses. By contrast, for many patients with PBC and PSC a diagnosis can now be made on the basis of biochemical, serological and/or radiological findings alone and histological confirmation may not be required. Liver biopsy can still be used to assess disease severity in such cases and remains important in establishing a diagnosis in patients with atypical features (e.g. AMA-negative PBC or the small-duct variant of PSC). Liver biopsy is also increasingly used in the assessment of patients suspected to have “overlap syndromes” involving AIH and PBC or PSC.     

肝移植后肝功能的异常

背景：肝移植后导致肝功能异常原因复杂,早期弄清引起肝功能异常的原因对治疗至关重要。 目的：较全面的了解肝移植后可以导致肝功能异常的原因,以便应用于临床诊治。 方法：应用计算机检索CNKI和FMJS数据库,采用医学主题词检索,检索词为“肝移植;肝功能异常;转氨酶异常;胆红素升高;原因”或“liver transplantation; abnormal liver function; transaminase abnormalities; bilirubin increased, and causes”,时间范围为1991年1月至2012年7月,共检索到98篇文章,选择文章主要内容与肝移植后肝功能异常直接相关的、发表在权威杂志上的文章共35篇进行综述。 结果与结论：肝移植后导致肝功能异常的原因众多,临床表现复杂。最常见的原因依次是急性排斥反应、胆道并发症及病毒感染。肝移植后早期,尤其是1个月内出现肝功能异常,需警惕小体积综合征和原发性移植物无功的发生。各种原因引起的肝功能异常,转氨酶及胆红素升高的程度不尽相同。急性排斥反应、自身免疫性肝炎、病毒感染、门静脉及肝静脉狭窄、缺血-再灌注损伤等转氨酶升高较胆红素升高显著;慢性排斥反应、胆道并发症、肝动脉狭窄、原发性胆汁性肝硬化、原发性硬化性胆管炎等早期以梗阻酶碱性磷酸酶、谷氨酰转移酶、总胆红素、直接胆红素升高为主;肿瘤导致的肝功能异常视肿瘤大小、压迫部位不同,可表现出以转氨酶升高为主或以胆红素升高为主。此外,各种原因多有其特殊病史,仔细询问病史有助于早期诊断。临床工作中,应重视尽量详尽的采集病史,根据转氨酶和胆红素升高的具体情况,首先考虑引起肝功能异常的常见原因,经临床证实排除后再考虑其他相对不常见原因,并结合实验室检查、影像学检查及肝脏穿刺病理活检,尽早明确病因及治疗。     

Transplanted liver: consequences of denervation for liver functions

Following liver transplantation, all hepatic nerves are transected; thus, liver allografts are completely isolated from neural control of their hosts. Despite this absolute denervation, liver allograft function does not appear to be significantly impaired after successful transplantation. In experimental animal models, hepatic denervation has no major effects on bile acid production and biotransformation, while it increases blood pressure and salt retention; decreases the number of hepatic progenitor cells, cholangiocyte proliferation, and liver regeneration; and influences the hepatic microcirculation, diet behavior, and glycemic control. In humans, hepatic denervation after liver transplantation has no major deleterious effects on bile secretion, liver regeneration, and hepatic blood flow. Insulin resistance and postprandial hyperglycemia, changes in ingestion behavior, and reduced stimulation of hepatic progenitor cells in the canals of Hering are the major side effects of absent liver innervation. Despite these abnormalities, patients can lead a new life with improved quality of life.     

自制铐式可调型肝钳在肝段切除中的应用

背景：研制自制铐式可调型肝钳以半肝血流阻断及肝局部血流阻断法行半肝、肝段及肝部分切除可减少肝功能损伤及胆瘘的发生。 目的：探讨自制铐式可调型肝钳以半肝血流阻断及肝局部血流阻断法行半肝、肝段及肝部分切除对血流变、肝肾功能的影响,评价其手术安全性。 方法：对85例半肝、肝段及肝部分切除的患者,应用自制铐式可调型肝钳以半肝血流阻断及肝局部血流阻断法行半肝、肝段及肝部分切除患者31例设为实验组,同期54例行不阻断肝门血流半肝、肝段及肝部分切除,比较术中出血量、术后胆瘘例数以及血流变学、肝肾功能的改变。 结果与结论：实验组31例患者术中出血量少、肝切除后无胆瘘、对血流变学、肝肾功能影响小。说明应用自制铐式可调型肝钳以半肝血流阻断及肝局部血流阻断法行半肝、肝段及肝部分切除,术式安全有效。     

Endotheliopathy of liver sinusoidal endothelial cells in liver disease

Liver is the largest solid organ in the abdominal cavity, with sinusoid occupying about half of its volume. Under liver disease, hemodynamics in the liver tissue dynamically change, resulting in injury to liver sinusoidal endothelial cells (LSECs). We discuss the injury of LSECs in liver diseases in this article. Generally, in noninflamed tissues, vascular endothelial cells maintain quiescence of circulating leukocytes, and unnecessary blood clotting is inhibited by multiple antithrombotic factors produced by the endothelial cells. In the setting of inflammation, injured endothelial cells lose these functions, defined as inflammatory endotheliopathy. In chronic hepatitis C, inflammatory endotheliopathy in LSECs contributes to platelet accumulation in the liver tissue, and the improvement of thrombocytopenia by splenectomy is attenuated in cases with severe hepatic inflammation. In COVID-19, LSEC endotheliopathy induced by interleukin (IL)-6 trans-signaling promotes neutrophil accumulation and platelet microthrombosis in the liver sinusoids, resulting in liver injury. IL-6 trans-signaling promotes the expression of intercellular adhesion molecule-1, chemokine (C-X-C motif) ligand (CXCL1), and CXCL2, which are the neutrophil chemotactic mediators, and P-selectin, E-selectin, and von Willebrand factor, which are involved in platelet adhesion to endothelial cells, in LSECs. Restoring LSECs function is important for ameliorating liver injury. Prevention of endotheliopathy is a potential therapeutic strategy in liver disease.     

去细胞化全肝生物支架修复肝损伤

BACKGROUND: Decellularized scaffolds are special for retaining the tubular structure used for nutrition transport, and providing a similar inner environment for cell growth. OBJECTIVE: To study the preparation of the decellularized whole liver bioscaffold and to explore its repair outcomes for liver injury. METHODS: Livers from 12 Sprague-Dawley rats were used for preparing the decellularized whole liver bioscaffold by chemical detergent-enzymes decellularized technology. Models of liver injury were established in another 24 Sprague-Dawley rats and randomized into two groups: the decellularized whole liver bioscaffold was implanted into the rat liver lesions in experimental group, and controls were given the injection of normal saline. Thirty days later, the serum levels of alanine aminotransferase and glutamic-oxaloacetic transaminase were detected, and liver tissues were removed for hematoxylin-eosin staining. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that extracellular matrix-like structures existed in the decellularized bioscaffold; cell components were completely removed from the liver, the collagen fibers in the scaffold arranged regularly and were not dissolved under electron microscope. The serum levels of alanine aminotransferase and glutamic-oxaloacetic transaminase in the experimental group were significantly lower than those in the control group (P < 0.05). Hematoxylin-eosin staining showed a large number of blue-stained and dense distributed nuclei, and pink distribution of collagen fibers that had no overt breakages in the control group, while pink and dense structures in the experimental group. These results suggest that the decellularized whole liver bioscaffold is easy to obtain, and can promote the injured liver repair.     

Human liver caudate lobe and liver segment

Recently, the caudate lobe has seemed to be the final target for aggresive cancer surgery of the liver. This lobe has five surfaces: the dorsal, left and hilar-free surfaces and the right and ventral-border planes. Surgeons have divided the caudate lobe into three parts: Spiegel’s lobe, which is called the ‘caudate lobe and papillary process’ by anatomists, the caudate process, viewed as almost the same entity by anatomists, and the paracaval portion corresponding to the dorsally located parenchyma in front of the inferior vena cava. All three parts are supplied by primary branches originating from the left and right portal veins, including the hilar bifurcation area. The hilar bifurcation branch often (50%) supplies the paracaval portion and it sometimes (29%) extends its territory to Spiegel’s lobe. It was postulated by Couinaud that the paracaval portion or the S9 is not defined by its supplying portal vein branch but by its ‘dorsal location’ in the liver. Couinaud’s caudate lobe or dorsal-liver concept caused, and still now causes, great logical confusion for surgeons. We attempt here to describe the margins of the lobe, border branches of the portal vein, the left/ right territorial border of the portal vein or Cantlie’s line and other topics closely relating to the surgery within these contexts. Finally, the caudate lobe as a liver segment will be discussed.