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。     

槲皮素及其衍生物金丝桃苷的抗肝纤维化机制研究进展

肝纤维化是一种常见的慢性进行性肝脏疾病,也是众多慢性肝脏疾病发展成为肝硬化或肝癌的必经阶段。在肝纤维化早期阶段采取有效的治疗措施,有可能阻止其向肝硬化或肝癌等方向进一步发展。深入探究肝纤维化的发病机制并寻找有效的肝纤维化防治靶点,具有重要的理论和临床意义。檞皮素及其衍生物金丝桃苷属于黄酮类化合物,具有抗纤维化、抗氧化应激、抗炎等多种生物活性功能,抗肝纤维化作用靶点丰富,作用通路复杂。本文从氧化应激损伤、炎症反应、肝星状细胞活化、细胞外基质沉积等方面对肝纤维化的发病机制展开回顾,旨在深入认识其可能的发病机制并寻找潜在的防治靶点。同时,对檞皮素及其衍生物金丝桃苷的研究进行综述,以期阐明其抗肝纤维化的作用机制,为防治肝纤维化的药物研发与临床治疗提供一定借鉴。     

柴芪益肝颗粒防治四氯化碳诱导大鼠肝纤维化的作用

目的观察柴芪益肝颗粒治疗四氯化碳诱导的大鼠肝纤维化的疗效及其对Leptin、TGF—β1和IL-13因子的影响。方法将50只SD大鼠随机分为模型组、柴芪益肝颗粒组、复方鳖甲软肝片组、秋水仙碱组和空白对照组,每组10只。大鼠腹腔注射四氯化碳诱导肝纤维化,光镜观察肝组织病理学改变,放射免疫分析法检测肝纤四项,液相芯片检测血清细胞因子。结果与模型组比较,柴芪益肝颗粒组肝脏病理变化显著减轻,肝小叶结构基本清晰,细胞索、肝窦无明显异常;与模型组比较,透明质酸（P=0．001）、层粘连蛋白（P=0．005）和Ⅳ型胶原（P=0．000）水平显著降低,瘦素（P=0．012）、转化生长因子131（TGF-β1）（P=0．000）和白介素13（IL-13）（P=0．016）水平亦显著降低,差异均有统计学意义。结论柴芪益肝颗粒显著改善肝纤维化,可能与其通过降低瘦素、TGF—β1和IL-13水平,抑制细胞外基质沉积有关。     

大黄素对肝纤维化大鼠肺损伤的保护作用

 目的:探讨不同剂量大黄素对肝纤维化大鼠肺损伤的保护作用。方法:采用复合致病因素法（CCl 4、乙醇、高脂、高胆固醇和低胆碱）建立肝纤维化大鼠模型并以不同剂量（20 mg/kg和40 mg/kg）大黄素进行治疗。4周后,测定肝指数,检测血清内毒素、同型半胱氨酸、反映肝功能的指标白蛋白、天门冬氨酸氨基转移酶、丙氨酸氨基转移酶、总胆红素、总胆固醇、甘油三酯和肝纤维化指标透明质酸、层黏连蛋白、Ⅳ型胶原蛋白、Ⅲ型前胶原蛋白的含量,观察肝组织病理学改变; 测定肺指数,光镜下行肺组织病理学观察,检测肺组织匀浆肿瘤坏死因子α(TNF-α)、丙二醛(MDA)、一氧化氮(NO)和过氧亚硝基阴离子(ONOO-)含量。结果:大鼠肝纤维化模型复制成功,模型组大鼠肺指数明显增加,肺脏发生水肿、炎症反应,肺匀浆TNF-α、MDA、NO和ONOO-含量明显增加;大黄素治疗组肺指数较模型组下降,肺组织病理性损伤明显减轻,肺组织TNF-α、MDA、NO和ONOO-含量明显降低。结论:大黄素对肝纤维化大鼠的肺损伤具有一定的保护作用。     

三七皂苷R1保护四氯化碳诱导肝纤维化模型大鼠的作用

背景：前期研究发现,三七总皂苷对小鼠免疫性肝损伤具有一定的保护作用。 目的：探究三七皂苷R1对四氯化碳诱导的肝纤维化模型大鼠的治疗作用。 方法：用四氯化碳诱导SD雄性大鼠制备肝纤维化模型,给药组按照60 mg/kg的剂量给予30 g/L三七皂苷R1溶液, 1次/d,连续4周和6周。对照组及模型组给予同体积的生理盐水,采用苏木精-伊红染色及Masson染色观察肝脏组织结构和纤维化程度分期;反转录-定量聚合酶链反应(qRT-PCR)检测法检测Ⅰ型胶原、α-平滑肌激动蛋白和转化生长因子β1表达水平。实验方案经昆明医科大学动物实验伦理委员会批准(批准号为approval No. KMMU2018018)。 结果与结论：①肝组织病理学显示,与模型组相比,三七皂苷R1能显著减轻纤维增生程度;②与模型组相比,三七皂苷R1组Ⅰ型胶原、α-平滑肌激动蛋白和转化生长因子β1表达水平显著降低(P < 0.05),三七皂苷R1给药4周与6周组比较差异无显著性意义;③结果提示,三七皂苷R1对四氯化碳诱导的肝纤维化模型大鼠具有一定的治疗作用。 ORCID: 0000-0002-0755-1476(吴朕) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

ADSCs对糖尿病肾病大鼠肾纤维化的抑制作用

目的探讨脂肪来源间充质干细胞(ADSCs)对糖尿病肾病(DN)大鼠肾纤维化的抑制作用。方法原代分离培养SD大鼠的ADSCs,鉴定干细胞的相关特性。荧光标记ADSCs并经尾静脉注射入DN大鼠体内,检测肾功。观察ADSCs在肾组织的分布及肾组织形态。Western blotting检测肾组织纤维化相关蛋白及Wnt通路相关蛋白的表达。结果 ADSCs具有间充质干细胞的形态,高表达干细胞相关抗原;经尾静脉注射的ADSCs可归巢入DN大鼠的肾组织内,改善了肾功指标,减轻了肾组织病理变化,纤维化指标及Wnt通路相关蛋白表达量明显下降。结论 ADSCs抑制糖尿病肾病大鼠的肾纤维化,对糖尿病肾病有治疗作用。     

RhoA和肌球蛋白轻链在肝纤维化大鼠肝组织中的表达

目的观察Rho/Rho激酶信号转导通路关键信号分子RhoA和磷酸化肌球蛋白轻链[p-MLC(Thr18/Ser19)]在大鼠肝纤维化组织中的表达规律。方法HE及Masson三色染色观察肝病理组织学变化;用Western blot检测RhoA、p-MLC(Thr18/Ser19)蛋白的表达;RT-PCR方法检测RhoAmRNA的表达。结果随着肝纤维化的进展,RhoA、p-MLC(Thr18/Ser19)蛋白表达明显增加,RhoAmRNA基因表达也逐渐加强。RhoA和p-MLC(Thr18/Ser19)分别与α-平滑肌肌动蛋白(α-SMA)呈显著正相关。结论Rho/Rho激酶信号转导通路在肝纤维化形成过程中发生变化。     

四氯化碳诱导肝纤维化早期大鼠肝窦内皮细胞及基底膜形态改变的动态研究

 目的：探讨四氯化碳诱导肝纤维化早期大鼠肝窦毛细血管化的形成过程。方法：清洁级雄性SD大鼠采用随机数字表法随机分为2组：正常对照组（N组,6只）和肝纤维化模型组（M组,32只）。M组大鼠腹腔注射50%四氯化碳蓖麻油混合液, N组大鼠腹腔注射生理盐水,剂量为2 mL/kg,每周2次,共4周。分别于造模第3天、1周、2周和4周处死大鼠,HE染色和Masson染色观察肝脏组织炎症及纤维化的改变,透射电镜观察肝窦内皮细胞（LSECs）窗孔与基底膜（BM）的改变,免疫组织化学检测LSECs表面标志物CD31及基底膜成分IV型胶原（Col IV）和层黏连蛋白（LN）的改变。结果：HE及Masson染色显示四氯化碳造模4周早期肝纤维化已形成。肝组织透射电镜显示四氯化碳造模第3天后开始出现LSECs窗孔直径变小及数目减少,随着造模时间的延长,LSECs失窗孔现象逐步严重,至第4周时局部内皮下可见连续的基底膜。免疫组化染色显示LSECs表面标志物CD31表达随着LSECs窗孔数目的减少而逐渐增强;基底膜成分Col IV于造模第2周时表达开始显著增强并随着造模时间延长表达逐渐增强,LN于造模第4周时表达开始显著增强。结论：肝纤维化早期大鼠局部肝组织可见典型的肝窦毛细血管化形成;肝窦壁内LN沉积是肝窦毛细血管化时形成连续基底膜的关键因素。     

Danshensu-mediated protective effect against hepatic fibrosis induced by carbon tetrachloride in rats

The culprit of hepatic fibrosis (HF) is linked to suprathreshold deposition of collagen. Thus, collagen reduction by improved metabolism contributes to HF management. In this study, we aimed to investigate the hepatoprotective effects of Danshensu (DSS) against carbon tetrachloride (CCl₄)-induced HF rats. The results showed that DSS-administrated rats resulted in decreasing in hepatosomatic indexes, and lowering serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Meanwhile, the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) were increased, while the content of malonaldehyde (MDA) was lessened in liver tissue of DSS administration group. In addition, the pro-fibrotic markers of hydroxyproline (Hyp), type III procollagen (PCIII) and hyaluronic acid (HA) contents were decreased. Histopathological examination confirmed that the hepatotoxicity in CCl₄-injured rats was alleviated following the DSS administration. Furthermore, intrahepatic protein expressions of alpha-smooth muscle actin (α-SMA), phosphorylated JAK2 (p-JAK2) and phosphorylated STAT3 (p-STAT3) were effectively down-regulated, respectively. Overall, this work demonstrates that DSS played the protective effect against CCl₄-induced cytotoxicity in liver tissue, which the probable mechanism is associated with attenuation of lipid peroxidation, collagen accumulation and enhancement of anti-oxidative defense capability, as well as regulation of intrahepatic JAK/STAT pathway for maintaining collagenic homoeostasis.     

Effect of cholinergic denervation on hepatic fibrosis induced by carbon tetrachloride in rats

Various factors involved in the development of liver fibrosis, including hepatic stellate cells (HSCs), cholinergic nervous activity and fibrogenetic cytokines. The present study aims to investigate the role of cholinergic regulation in the promoting of liver fibrogenesis relating to bone morphogenetic protein-6 (BMP-6) and/or transforming growth factor-beta1 (TGFbeta1). We treated carbon tetrachloride (CCl(4)) into rats for eight weeks to induce liver fibrosis and arranged these rats for cholinergic denervation, hepatic branch vagotomy or atropine administration. Acetylcholinesterase (AChE) staining showed the distribution of cholinergic nerve around fibrosis scaring septa. The immunohistochemical staining for alpha smooth muscle actin (alphaSMA) indicated the less HSCs in CCl(4) treated rat liver with cholinergic denervation as compared to the sham-operated CCl(4) treated rats. It seems that cholinergic nerve not only innervates around the fibrosis area but also promotes HSCs. We also detected TGFbeta1 and BMP-6 expressions using RT-PCR and immunohistochemistry. The obtained results show that cholinergic denerveration decreases BMP-6 and TGF-beta1 expressions in CCl(4) induced liver fibrosis of rats. In conclusion, cholinergic nerve may influence HSCs in addition to the lowering of BMP-6 and TGF-beta1 gene expressions to modify liver fibrosis.     

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。     

The effects of grape seed and colchicine on carbon tetrachloride induced hepatic damage in rats

This study aims to determine the effects of grape seed and colchicine on carbon tetrachloride (CCl₄) induced hepatic damage and on some serum biochemical parameters. Sixty male Wistar albino rats (200–250 g) were randomly divided into six groups (ten rats/group) and included the control group the group were given isotonic sodium chloride (1 mL/kg b.w) intraperitonealy (i.p.), group 2 the group treated i.p. injection of CCl₄ (1.0 mL/kg b.w) in corn oil twice in the first week, Groups 3 and 4 injected with CCl₄ as described for group 2 and the rats were orally given (100 mg/kg b.w) GSE and i.p. injected (10 μg/rat) with colchicine for four weeks, respectively and groups 5 and 6 were the grape seed and colchicine control groups in which rats were orally given grape seed (100 mg/kg b.w) and i.p. injected with colchicine (10 μg/rat), respectively. Anorexia, weight loss, motionlessness and hepatic colour variation at necropsy were observed in groups 2, 3, and 4. Hyperemia, focal bleeding, fat degeneration, changes ranging from degenerative to necrotic, increase in connective tissue elements, pronounced in portal sites in particular, and infiltration of lymphoid series cell observed in the livers of the rats in group 2, treated with CCl₄. Histological hepatic changes in the rats in group 3 and 4 were similar to those in group 2. The levels of serum total protein, albumin and globulin decreased in groups 2, 3, and 4, compared with groups 1, 5 and 6; aspartate transaminase (ALT) activities increased. The lowest alkaline phosphatase (ALP) activities were in groups 4 and 5. We concluded that GSE and colchicine have not sufficient ameliorative effects to CCl₄ induced acute hepatic damage.     

Liver precursor cells increase hepatic fibrosis induced by chronic carbon tetrachloride intoxication in rats

Hepatic fibrosis, the major complication of virtually all types of chronic liver damage, usually begins in portal areas, and its severity has been correlated to liver progenitor cells (LPC) expansion from periportal areas, even if the primary targets of injury are intralobular hepatocytes. The aim of this study was to determine the potential fibrogenic role of LPC, using a new experimental model in which rat liver fibrosis was induced by chronic carbon tetrachloride (CCl(4)) administration for 6 weeks, in combination with chronic acetylaminofluorene treatment (AAF), which promotes activation of LPC compartment. Treatment with CCl(4) alone caused a significant increase in serum transaminase activity as well as liver fibrosis initiating around central veins and leading to formation of incomplete centro-central septa with sparse fibrogenic cells expressing α-smooth muscle actin (αSMA). In AAF/CCl(4)-treated animals, the fibrogenic response was profoundly worsened, with formation of multiple porto-central bridging septa leading to cirrhosis, whereas hepatocellular necrosis and inflammation were similar to those observed in CCl(4)-treated animals. Enhanced fibrosis in AAF/CCl(4) group was accompanied by ductule forming LPC expanding from portal areas, αSMA-positive cells accumulation in the fibrotic areas and increased expression of hepatic collagen type 1, 3 and 4 mRNA. Moreover, CK19-positive LPC expressed the most potent fibrogenic cytokine transforming growth factor-β (TGFβ) without any expression of αSMA, desmin or fibroblast-specific protein-1, demonstrating that LPC did not undergo an epithelial-mesenchymal transition. In this new experimental model, LPC, by expressing TGFβ, contributed to the accumulation of αSMA-positive myofibroblasts in the ductular reaction leading to enhanced fibrosis but also to disease progression and to a fibrotic pattern similar to that observed in humans.     

肝星状细胞SCAP特异性敲除对小鼠肝纤维化的影响

目的:探讨固醇调节元件结合蛋白裂解活化蛋白(SCAP)在肝星状细胞中缺失能否延缓小鼠肝纤维化进程.方法:将SCAPloxP/loxP小鼠与Lrat-Cre工具鼠繁殖得到Lrat-Cre+/+SCAPfl/fl、Lrat-Cre+/?SCAPfl/fl和Lrat-Cre?/?SCAPfl/fl小鼠,并用PCR法对小鼠基因...