家兔肝纤维化时肝组织RNA含量变化

家兔肝纤维化时肝组织ＲＮＡ含量变化彭雁忠，黄其通，邓斌，胡积军（湖北医大附属第一医院，武汉４３００６０）正常猪肝的ＲＮＡ具有阻抑肝纤维化作用，本文观察了血吸虫肝纤维化时ＲＮＡ含量变化。实验用家兔７２只，体重２．０～２．５ｋｇ，同龄。随机分为正常组、病...     

RNA治疗兔肝纤维化后肝组织RNA含量变化

ＲＮＡ治疗兔肝纤维化后肝组织ＲＮＡ含量变化△彭雁忠邓斌杨清杨丽华黄其通（湖北医科大学附一院，武汉４３００６０）肝纤维化时肝组织ＲＮＡ含量下降，ＲＮＡ具有抗肝纤维化的作用。本实验旨在探讨从体外补充ＲＮＡ能否提高实验动物肝脏ＲＮＡ含量且具有抗肝纤维化作用...     

RNA和马洛替酯治疗家兔肝纤维化后肝组织形态学变化

ＲＮＡ和马洛替酯治疗家兔肝纤维化后肝组织形态学变化１胡积军２彭雁忠２邓斌２杨丽华２黄其通２有人报告［１］核糖核酸（ＲＮＡ）有抗肝纤维化作用，血吸虫肝纤维化发展到一定阶段肝组织ＲＮＡ含量逐渐减少［２］。本实验选用ＲＮＡ和马洛替酯来阻抑实验性血吸虫肝纤维...     

微小RNA与心房颤动心房纤维化

心房纤维化是心房颤动(房颤)发生与维持的重要因素,微小RNA(microRNA,miRNA)通过调控纤维化相关靶基因,在房颤心房纤维化中起着重要作用。因此,深入研究miRNA在心房纤维化中的作用机制将为房颤的诊治提供新的思路。     

微小RNA与器官纤维化研究进展

微小RNA（miRNA）是一类长21～25个核苷酸的内源性非编码小RNA,在转录后水平调控靶基因的表达。近年来研究发现,miRNA也参与了机体多个器官如心脏、肺脏、肾脏、肝脏、皮肤的纤维化过程。本文对miRNA生物学特征及其在脏器纤维化过程中的作用的相关研究进展进行综述。     

gp73在小鼠肝纤维化肝组织中的表达及机制

目的在小鼠肝组织中探讨高尔基体跨膜糖蛋白73(gp73)在肝纤维化进程中的变化及机制。方法腹腔注射CCl4构建C57BL/6J小鼠肝纤维化模型;用ELISA检测小鼠血清中gp73的水平;用免疫组织化学染色检测肝组织内gp73的表达;用密度梯度离心法分离肝星状细胞(HSC)并检测其中gp73的表达;用免疫组织化学染色法检测肝组织中胰岛素样生长因子2 mRNA结合蛋白3(IGF2BP3)的表达;接下来分别转染IGF2BP3过表达与敲低的质粒,用RT-PCR检测GP73蛋白的编码基因GOLM1的表达变化;流式细胞计量术检测基因敲除鼠肝纤维化模型中HSC内gp73的表达。结果在肝纤维化模型的小鼠血清(P<0.01)、肝组织(P<0.05)及原代HSC(P<0.001)中gp73蛋白表达水平均升高,同时组织中的IGF2BP3蛋白表达也升高(P<0.05)。细胞系中转入IGF2BP3的过表达及敲低质粒后,成功过表达IGF2BP3(P<0.001)和敲低(P<0.01),GOLM1的表达也随之上调(P<0.001)和下调(P<0.01)。随后在IGF2BP3敲除鼠的肝纤维化模型的HSC中检测发现gp73表达降低(P<0.01)。结论在CCl4诱导的小鼠肝纤维化模型中,由HSC表达的gp73蛋白水平升高,而RNA结合蛋白IGF2BP3是促进其表达的潜在调控因素。     

卵巢切除对四氯化碳诱导大鼠肝纤维化形成的影响

为探讨卵巢切除对CCl4 诱导大鼠肝纤维化形成的影响 ,采用CCl4 诱导雌性大鼠肝纤维化动物模型 ,观察卵巢切除及雌激素替代治疗 (苯甲酸雌二醇 1mg kg)对肝脏胶原沉积和I、Ⅲ型胶原蛋白表达的影响 ,并分别检测血清学标志及肝脏组织学等变化。结果显示CCl4 模型组大鼠肝脏发生典型的肝纤维化改变 ,卵巢切除组的肝脏胶原沉积更为明显 ,肝脏表达I、Ⅲ型胶原及血清肝纤维化指标也明显高于CCl4 摸型组 (P <0 0 5 ) ,而雌激素干预及替代治疗则可抑制肝纤维化的形成。表明卵巢切除加速CCl4 诱导大鼠肝纤维化的形成 ,其发生可能与卵巢分泌的雌激素对肝纤维化的抑制作用有关。     

卵巢癌组织与正常卵巢组织差异表达谱的初步分析

目的:通过采用限制片段差异显示聚合酶链反应(RFDD-PCR)建立卵巢癌组织与正常卵巢组织基因差异表达谱,分析比较卵巢癌发病机制相关基因的表达差异。方法:采集上皮性卵巢癌手术患者的卵巢癌组织和正常卵巢组织,用RFDD-PCR技术平行检测,对各组织间有表达差异的基因进行生物信息学分析,筛选与卵巢癌发病机制相关的基因。结果:运用RFDD-PCR技术,构建了两种不同组织的基因表达谱,通过对卵巢癌组织及正常卵巢组织差异表达谱的分析中发现与肿瘤的发生、发展密切相关的原癌基因和抑癌基因、细胞周期相关基因、增殖和凋亡相关基因、血管生长因子、代谢酶等6类基因36种因子的表达差异。结论:利用RFDD-PCR差异显示技术能够建立卵巢癌组织和正常卵巢组织基因差异表达谱,是对肿瘤的发病机制和寻找治疗靶点研究的一种有效的手段。     

苦瓜对实验性大鼠肝纤维化的干预作用及可能机制

目的: 探讨苦瓜(BM)对四氯化碳(CCl₄)诱导大鼠肝纤维化的干预作用及相关机制。方法: 随机将32只雄性健康Wistar大鼠分为4组:对照组(C组);模型组(CCl₄,M组);BM低剂量组(BM 100g/kg饲料+CCl₄,BM-L组)、BM高剂量组(BM 200g/kg饲料+CCl₄,BM-H组)。饲养中除C组外的各组大鼠均皮下注射50%CCl₄-橄榄油溶液2 mL/kg,2次/周,共8周,诱导肝纤维化动物模型。8周后处死大鼠,留取大鼠肝脏和血清。计算肝体指数;测定血清丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;测定肝匀浆总蛋白(TP)和白蛋白(Alb)含量、谷胱甘肽过氧化物酶(GSH-Px)活性、羟脯氨酸(HYP)含量和单胺氧化酶(MAO)活性;胶原纤维染色观察大鼠肝组织变性与胶原沉积病理改变。结果: 与M组比较,摄入BM后的各剂量组大鼠肝体指数显著降低(P<0.01);血清MDA含量及肝匀浆HYP含量和MAO活性均明显降低(P<0.01),而血清SOD活性、肝组织TP和Alb含量、GSH-Px活性明显增强(P<0.01)。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;BM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,以高剂量组更明显。结论: BM具有抗CCl₄诱导大鼠肝纤维化作用,其机制可能与其抗脂质过氧化、降低肝HYP含量及MAO活性的作用有关。     

人参总甙抗四氯化碳损伤大鼠离体肝细胞的作用

本实验采用大鼠离体肝细胞原代培养２４ｈ并利用ＣＣｌ４造成急性细胞损伤模型，检定人参总甙（ＧＳＳＤ）对肝细胞损伤的影响。结果表明：ＧＳＳＤ可显著降低中毒肝细胞的脂质过氧化物水平。抑制肝细胞脂质过氧化，并降低谷丙转氨酶（ＧＰＴ）和谷草转氨酶（ＧＯＴ）水平，保护脂质膜。ＧＳＳＤ显著促进中毒肝细胞ＲＮＡ和ＤＮＡ的合成，超微结构观察证实ＧＳＳＤ能减轻ＣＣｌ４对肝细胞染色质，线粒体，内质网和核蛋白体的损害，结     

芍甘多苷对四氯化碳亚急性肝损伤大鼠肝脏基因表达谱的影响

目的 研究芍药甘草汤提取物芍甘多苷对四氯化碳(CC14)亚急性肝损伤大鼠血清转氨酶、肝脏病理及肝脏基因表达谱的影响.方法 60只SD大鼠完全随机分6组(n=10),即正常组、模型组、阳性对照组(联苯双酯100 mg/kg)和芍甘多苷低、中、高3个剂量组(88、264、528 mg/kg).采用CCl4诱发大鼠亚急性肝损伤,然后给予不同剂量的芍甘多苷灌胃,观察药物对血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、肝脏组织病理的影响;并运用基因表达谱芯片技术,研究其对肝脏基因表达谱的影响.结果 与模型组比较,芍甘多苷高、中剂量灌胃给药均能显著降低CCl4亚急性肝损伤大鼠血清ALT[ (218.37± 114.59,234.59± 109.33) U/L比(389.12±142.57) U/L,P＜0.05];且高剂量芍甘多苷可以显著降低肝脏组织学评分(2.42±0.52比3.92±0.84,P＜0.05).芍甘多苷能上调和(或)下调与肝损伤相关的多种基因的表达.结论 芍甘多苷有明显的肝脏保护作用,其机制与调节与肝损伤相关的多种基因的表达有关,是多靶点作用药物.     

Dasatinib prevent hepatic fibrosis induced by carbon tetrachloride (CCl4) via anti-inflammatory and antioxidant mechanism

Objectives: Dasatinib, a potent and broad-spectrum tyrosine kinase inhibitor, is approved for the treatment of imatinib-resistant chronic myelogenous leukemia. The aim of this study was to evaluate the anti-fibrotic, anti-inflammatory and antioxidant effects of this agent against CCl₄-induced hepatic fibrosis and oxidative status.

Materials and methods: Experimental fibrosis was induced in Wistar male rats by 12 weeks of CCl₄ administration (i.p.). During the last 8 weeks of injection, rats were gavaged daily with Dasatinib (10?mg/kg). To evaluate anti-inflammatory and anti-fibrotic effects of Dasatinib, histopathological examination of liver tissue was performed and serum ALT and AST activities, oxidant, antioxidant parameters and hepatic tumor necrosis factor alpha (TNF-α) were examined. Moreover, transforming growth factor (TGF-β₁), platelet derived growth factor (PDGF) and TNF-α mRNA expressions were also evaluated by real time polymerase chain reaction.

Results: Dasatinib administration induced a significant reduction of ALT and AST activities (p?4 injected rats (p?1 and PDGF were increased due to CCl₄ intoxication (p?p?p?4 administration which was significantly attenuated by Dasatinib (p?Discussion and conclusion: Our findings indicate that Dasatinib can be cautiously an anti-fibrotic, anti-inflammatory and anti-oxidative agent in clinical setting.     

Establishment of a liver fibrosis model in cynomolgus monkeys

Hepatic fibrosis, resulted from hepatoxicity and other diseases such as diabetes, is an important pathological characteristic of chronic liver diseases. Establishment of hepatic fibrosis animal models is of great importance and a prerequisite for human clinical studies. The common models for liver fibrosis are often established in lower small animals such as rats, but non-human primates are a much better model for human diseases because of the physiological similarity with humans. In this study, we investigated the method to induce liver fibrosis in cynomolgus monkeys using carbon tetrachloride (CCl₄) and to establish a model that more closely mimics human liver fibrosis. We successfully established the liver fibrosis model in 15 of the 20 cynomolgus monkeys (success rate 75%), by CCl₄ administration at a dose of 1.0 mL/kg (400 mL/L) twice a week. Liver biopsy showed that liver fibrosis progressed with time and gradually advanced into early-stage cirrhosis in 10 of the 15 established models at 16 weeks. Our study provides a better research platform for the prevention and treatment of chronic liver diseases.     

钙中性蛋白酶2及Bax在肝纤维化大鼠肝组织中的表达变化及意义

 目的：探讨钙中性蛋白酶2（calpain 2）及Bcl-2相关X蛋白(Bax)在肝纤维化过程中的表达变化及其在肝纤维化过程中的可能作用。方法:  雄性Wistar大鼠40只,随机分为正常对照4周、8周组和肝纤维化模型4周、8周组,每组各10只。肝纤维化组按3 mL/kg体重的剂量皮下注射40% CCl4植物油溶液,每隔3 d注射1次,造模时间分别为4周和8周;对照组大鼠皮下注射等量植物油溶液。采用TUNEL法检测肝组织中肝细胞凋亡情况;real-time PCR检测肝组织中calpain 2及bax mRNA的表达变化。免疫组织化学法及Western blotting检测肝组织中calpain 2及Bax蛋白的表达情况。 结果:  Real-time PCR检测发现肝纤维化4周和8周组大鼠肝组织中calpain 2及bax mRNA表达较相应的正常对照组显著增加。免疫组化及Western blotting检测显示肝纤维化4周组大鼠肝组织中calpain 2蛋白的表达与正常4周组比较无显著差异;随着肝纤维化程度的加重,肝纤维化8周时大鼠肝组织中calpain 2的表达显著增加;而肝组织中Bax的表达从肝纤维化4周时就显著增加,肝纤维化8周时达到高峰。此外,通过TUNEL法检测发现肝纤维化4周和8周组大鼠肝组织中肝细胞凋亡的数目较正常组大鼠显著增加。结论:  Calpain 2与Bax可能参与了肝纤维化的发生发展过程。     

山丹黄参多糖对四氯化碳诱导的慢性肝损伤的保护作用

目的 探讨山丹黄参多糖对四氯化碳（CCl4）小鼠肝损伤的保护作用。方法 将40只小鼠分为正常对照组、实验对照组、山丹黄参多糖1～3组5个组,山丹黄参多糖1～3组分别连续灌胃浓度为12.5、25.0和37.5 g/L的山丹黄参多糖（共28 d,每天2次,每次 0.2ml）,正常对照组和实验对照组灌胃等量的生理盐水,灌胃多糖第3天起,实验对照组：山丹黄参多糖1、2、3组各组小鼠于下午灌胃 2 h后分别腹腔注射0.2％CCl4橄榄油溶液 0.2ml（每3天1次,共9次）,正常对照组腹腔注射等量的生理盐水。用光学显微镜观察肝组织的结构变化,用比色法检测血浆谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)和肝组织超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量的变化,用免疫组织化学法检测Caspase-3及Bax蛋白在肝组织表达的变化。 结果 实验对照组血浆ALT、AST和ALP含量显著升高（P<0.01）,肝组织SOD活力明显降低（P<0.01）,MDA含量显著升高（P<0.01）,肝组织结构不清,肝细胞炎性坏死,空泡化严重,Caspase-3蛋白和Bax蛋白阳性表达明显增强（P<0.01）。山丹黄参多糖各组血浆ALT、AST、ALP活性明显降低,肝组织SOD活力显著升高（P<0.01）,MDA含量明显下降（P<0.01）,肝索清晰,炎性坏死减少,肝细胞结构清晰,Caspase-3及Bax蛋白阳性表达明显减少（P<0.01）。结论 山丹黄参多糖可提高细胞抗氧化活性,具有抗炎、抗氧化、抑制细胞凋亡和保护肝细胞等作用,能有效减轻CCl4对肝组织的损伤,且在一定范围内呈剂量效应。     

Changes in microRNA expression induced by rabies virus infection in mouse brains

MicroRNAs (miRNAs) are small RNA (≈22 nt) molecules expressed endogenously in cells. They are involved in the regulation of gene expression. Recently, evidence has shown that cellular miRNAs have key regulatory roles in virus-host interactions. The rabies virus (RABV) causes a fatal infection of the central nervous systems (CNS) of warm-blooded animals, yet its pathogenesis remains poorly understood. To gain more insight into the pathogenesis of RABV, a miRNA microarray was performed as part of an investigation of changes in host miRNA expression in the brains of mice infected with RABV. The results showed that RABV infection induced modulation of the expression of sixteen miRNA molecules. These data were verified by real-time PCR. Functional analysis showed the differentially expressed miRNAs to be involved in many immune-related signaling pathways, such as the RIG-I-like receptor signaling pathway, JAK-STAT signaling pathway, chemokine signaling pathway, T-cell receptor signaling pathway, MAPK signaling pathway, leukocyte transendothelial migration, and natural killer cell mediated cytotoxicity. The predicted expression levels of the target genes of these modulated miRNAs correlated with measurements of gene expression measured by DNA microarray and qRT-PCR.     

四氯化碳诱导的肝损伤对大鼠免疫机能的影响

目的:研究化学性肝损伤对机体免疫机能的影响。方法:选择30只健康、1月龄、平均体重(83.20±2.11) g雌雄各半SD大鼠,经一段时间适应性饲养后,随机分成四氯化碳(CCl₄)注射组和对照组。CCl₄注射组大鼠按0.2 mL/100 g体重剂量每周定时注射CCl₄ 2次, 连续注射8周;对照组同时注射等量的橄 榄油。观察CCl₄诱导的肝损伤对机体免疫功能的影响。结果:注射CCl₄大鼠外周血T淋巴细胞比例和白细胞介素2水平显著低于对照组,与此同时B淋巴细胞比例却显著高于对照组;注射CCl₄组大鼠血浆总蛋白含量和白蛋白、α和β球蛋白比例下降,而γ球蛋白上升。结论:CCl₄诱导的肝损伤能明显影响大鼠的细胞免疫和体液免疫功能,造成大鼠的免疫功能失调。     

Blockade of CCN4 attenuates CCl4-induced liver fibrosis

Introduction

CCN4, also termed WNT-inducible signaling pathway protein-1 (WISP-1), has important roles in inflammation and tissue injury. This study aimed to investigate the effect of CCN4 inhibition using monoclonal anti-CCN4 antibody (CCN4mAb) on the liver injury and fibrosis in a mouse model of liver fibrosis.

Material and methods

The mouse liver fibrosis model was induced by carbon tetrachloride (CCl₄). Mice received vehicle (saline/olive oil) by subcutaneous injection, CCl₄ by subcutaneous injection or CCl₄ (subcutaneous) plus CCN4mAb by subcutaneous injection. The pro-inflammatory and pro-fibrotic factors were determined by Western blot. The biochemistry and histopathology, collagen deposition and nuclear factor (NF)-κB activity were also assessed.

Results

Chronic CCl₄ treatment caused liver injury and collagen accumulation. The expression levels of CCN4, pro-inflammatory and pro-fibrotic mediators as well as the activity of NF-κB were markedly increased. Treatment with CCN4mAb significantly inhibited CCl₄-induced CCN4 expression, leading to attenuated CCl₄-induced liver injury and the inflammatory response. CCN4 blockade also significantly reduced the formation of collagen in the liver and the expression of α-smooth muscle actin and transforming growth factor β1.

Conclusions

CCN4 inhibition by CCN4mAb in vivo significantly attenuated the CCl₄-induced liver injury and the progression of liver fibrosis. CCN4 may represent a novel therapeutic target for liver injury and fibrosis.