共查询到19条相似文献，搜索用时 125 毫秒
1.
皱纹盘鲍多糖抗肿瘤药理作用研究 总被引：6，自引：0，他引：6
对从软体动物皱纹盘鲍ＨａｌｉｏｔｉｓｄｉｓｃｕｓｈａｎｎａｉＩｎｏ中分离提取的纯多糖进行了抗肿瘤活性研究。药理实验表明,多糖对体外培养的小鼠移植性肿瘤肉瘤Ｓ１８０,肝癌ＨｅｐＡ细胞无细胞毒作用,但能明显延长ＨｅｐＡ小鼠的生存时间,抑制小鼠移植性肉瘤Ｓ１８０的生长,在抑瘤的同时,能明显提高免疫器官胸腺,脾脏的重量,降低荷瘤小鼠肝脏中过氧化脂质的水平,提示鲍鱼多糖可能是通过增强荷瘤小鼠的免疫功能和抗氧 相似文献
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目的：观察参苓白术散对非酒精性脂肪性肝病（NAFLD）大鼠肝细胞、Kupffer细胞、哺乳动物雷帕霉素靶蛋白（mTOR）通路相关基因及蛋白表达的影响,以阐明参苓白术散抗NAFLD的作用机制。方法：将80只大鼠随机分为正常组,模型组及参苓白术散低、高剂量组。对模型组、各药物组连续8周喂养高脂饲料制备NAFLD大鼠模型。造模成功后采用油红O染色观察肝组织病理变化;Ⅳ型胶原酶离体循环灌注法分离肝细胞、Kupfer细胞;全自动生化仪检测血清总胆固醇（TC）、三酰甘油（TG）;酶联免疫吸附法测定肝细胞及Kupffer细胞血管内皮生长因子（VEGF）、干扰素（IFN）-γ含量;实时荧光定量PCR及蛋白质印迹法（Western blot）检测大鼠肝细胞及Kupffer细胞脑内富含的小G蛋白Ras同系物（Rheb）、mTOR复合物1（mTORC1）、核糖体S6蛋白激酶1（S6K1）和真核细胞翻译起始因子4E-结合蛋白1（4E-BP1）mRNA和蛋白表达。结果：模型组大鼠肝组织存在严重的脂质蓄积。血清TC、TG,肝细胞、Kupffer细胞,VEGF、IFN-γ含量,以及Rheb、mTORC1、S6K1、4E-BP1 mRNA和蛋白表达水平较正常组显著升高（P<0.01）,说明造模成功。与模型组比较,各药物干预组血清TC、TG,肝细胞、Kupffer细胞,VEGF、IFN-γ含量,以及Rheb、mTORC1、S6K1、4E-BP1 mRNA和蛋白表达均有不同程度的下调（P<0.05或P<0.01）。其中参苓白术散高剂量组下调趋势最为显著,与低剂量组比较,差异均有统计学意义（P<0.05或P<0.01）。结论：参苓白术散抗NAFLD的作用机制可能与抑制肝细胞及Kupffer细胞mTOR通路中Rheb、mTORC1、S6K1、4E-BP1基因及蛋白表达有关。 相似文献
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螺旋藻及其多糖、多糖蛋白提取物对体外癌细胞的抑制作用 总被引：13，自引：0，他引：13
螺旋藻多糖是螺旋藻的重要生物学活性成分,１９９１年刘力生等证明它对腹水型肝癌细胞及肉瘤１８０有杀伤抑制作用［１］;１９９６年日本科学家证明它有抗病毒及免疫力提高作用。本研究用热水浸提法提取螺旋藻多糖及多糖蛋白,并测定它对肺癌细胞,人白血病淋巴细胞及胃癌细胞的杀伤抑制作用。１材料及方法１．１样品及试剂来源钝顶螺旋藻（Ｓｐｉｒｕｌｉｎａｐｌａｔｅｎｓｉｓ）、极大螺旋藻（Ｓ．ｍａｘｉｕｍ）及螺旋藻干粉由云南程海宏源生物技术公司提供,部分藻粉由本实验室自制。昆明种小鼠,重３０～４０ｇ,由昆明医学院提供。胃癌… 相似文献
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大菱鲆鳍细胞系的建立 总被引：2，自引：1，他引：2
建立大菱鲆(Scophthalus maximus)鳍细胞系,文中采用胰蛋白酶、透明质酸酶和Ⅱ型胶原酶消化法启动大菱鲆鳍组织的原代培养,并通过培养液配方和培养条件的优化成功进行大菱鲆鳍细胞的继代培养。研究结果显示,在pH=7.0~7.4、温度20~24℃的培养条件下,培养于含有表皮细胞生长因子、碱性成纤维细胞生长因子、羧甲基壳多糖、N-乙酰葡萄糖盐酸盐的Leibovitz-15培养液(20%胎牛血清)中的大菱鲆鳍细胞,其生长分裂状况最好,细胞形态为成纤维细胞样。经继代培养后,细胞生长分裂依然十分旺盛,第60代大菱鲆鳍细胞系的群体倍增时间为62.4 h,虽然出现了染色体的非整倍性,但其特征性染色体仍为44条。该细胞系细胞经液氮冻存后仍保持有原有形态和较高存活率。现已成功建立了连续性大菱鲆鳍细胞系,目前已传至第65代。该细胞系的建立对于查清病毒对大菱鲆细胞的感染途径与感染机理等具有重要的理论意义,对于病毒疫苗研制具有重要应用价值。 相似文献
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由青岛沿岸潮间带表层沉积物分离出４株光合细菌，经鉴定属于荚膜红假单胞菌（Ｐｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｃａｐｓｕｌａｔａ）（菌株ＨＤ３、ＲＳ、ＭＤ２）和球形红假单胞菌（Ｒｈｏｄｏｐｓｅｕｄｏｍｏｎａｓｓｐｈｅｒｏｉｄｅｓ）（菌株ＰＳ２－２）。光照厌氧培养生长实验表明ＨＤ３的最适盐度为Ｓ＝１５～２５，ＰＳ２－２为一淡水菌株。菌株ＨＤ３的最适ＰＨ范围为６．３５～８．３。ＨＤ３、ＲＳ均不能利用硫化物且其生长受较高浓度的硫化物抑制。 相似文献
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目的：探讨肝宁方对非酒精性脂肪性肝炎（NASH）小鼠的保护作用及其对活化后大鼠肝星状细胞（HSCs）的影响。方法：将24只小鼠随机分为正常组、模型组、肝宁方组,每组各8只。对模型组、肝宁方组采用高脂饮食联合四氯化碳豆油溶液皮下注射建立NASH小鼠模型,造模成功后正常组和模型组予0.9%氯化钠注射液灌胃,肝宁方组予肝宁方灌胃。干预3周。将大鼠HSCs分成正常组、模型组及肝宁方组,正常组予正常培养基培养,模型组及肝宁方组采用转化生长因子-β1（TGF-β1）活化,随后正常组和模型组用含正常大鼠血清培养基培养,肝宁方组加入含30%肝宁方含药血清培养基培养。培养24 h。观察肝组织病理学情况;血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、血清总胆固醇（CHOL）、三酰甘油（TG）水平;肝组织中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）表达,葡萄糖调节蛋白78（GRP78）及内质网核信号转导蛋白1α（IRE-1α）蛋白表达;肝细胞凋亡情况;HSCs的增殖情况。结果：肝宁方组和模型组小鼠相比,脂质空泡和出血减少,炎性细胞浸润减轻。小鼠血清AST、ALT、CHOL、TG水平,肝组织TNF-α、IL-6水平,GRP78、IRE-1α蛋白表达,肝细胞凋亡情况模型组与正常组比较,肝宁方组与模型组比较,差异均有统计学意义（P<0.01或P<0.05）。与模型组相比,肝宁方含药血清干预后HSCs增殖趋势明显减弱（P<0.01）。结论：肝宁方能够改善NASH小鼠肝脏病理学变化,保护肝脏、调节脂质代谢、抑制炎症反应、减轻肝细胞凋亡、抑制HSCs增殖,其机制可能与调控内质网应激有关。 相似文献
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褐点石斑鱼三种组织细胞系的建立 总被引：2，自引：1，他引：1
为建立褐点石斑鱼鳍、心脏和鳔组织细胞系,本文利用不同培养液和培养温度对褐点石斑鱼鳍、心脏和鳔组织细胞进行了原代培养和传代培养。实验结果显示,在24℃培养于含有羧甲基壳寡糖、碱性成纤维样生长因子、I型胰岛素样生长因子及20%胎牛血清的DMEM/F12培养液(pH=7.2)中的3种细胞,均为成纤维样细胞,其生长分裂状态最佳,可以持续稳定传代。第60代褐点石斑鱼鳍、心脏和鳔细胞的群体倍增时间分别为50.6 h、40.3 h和43.3 h,其特征性染色体数目均为48条。目前鳍、心脏和鳔细胞已分别传至第90代、第70代和第75代,已成功建立了3种细胞的连续性细胞系,为鱼类病毒与宿主细胞相互作用机制等鱼类病毒学基础研究,以及鱼类病毒的分离、鉴定、繁殖及病毒疫苗研制等奠定了基础。 相似文献
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螺旋藻多糖对CD3AK细胞增殖能力的影响 总被引：7，自引：0，他引：7
研究了螺旋藻多糖（ＰＳ）对ＣＤ３ＭｃＡｂ激活的杀伤细胞（ＣＤ３ＭｃＡｂ Ａｃｔｉｖａｔｅｄ Ｋｉｌｌｅｒ Ｃｅｌｌｓ,ＣＤ３ＡＫ Ｃｅｌｌｓ）增殖能力的影响。结果表明,当ＰＳ浓度为２．５μｇ·ｍｌ＾－１培养体系条件下,对ＣＤ３ＡＫ细胞具有明显的刺激细胞增殖作用（Ｐ〈０．０２）;对培养长达２３ｄ的ＣＤ３ＡＫ细胞杀伤肿瘤细胞（Ｋ５６２细胞）的活性仍维持在较高的水平（４６．５％￣５０％）。提示ＰＳ对辐射 相似文献
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An isolated fish hepatocyte culture system was developed as a models ystem to investigate the mechanisms of action of environmental contaminants. Hepatocytes were isolated from striped bass (Morone saxatilis) by an adaptation of the two stage perfusion technique of Seglen.1 The system was used to evaluate metal binding protein (MBP) induction in response to cadmium, a primary inducer of metallothionein (MT) in rat hepatocytes. Striped bass hepatocytes appeared to be refractory to the induction of MBP by cadmium, since there was no significant increase in the synthesis of MBP for any of the doses at any of the time points investigated. However, when a similar experiment was performed using rat hepatocytes there was induction of MBP that was related to both dose and time. These comparative experiments indicate that although there are similarities between the hepatocytes of the two species in regard to 35S incorporation into low molecular weight metal-binding proteins, there appear to be significant quantitative differences as well in regard to MBP kinetics. This in vitro model system could potentially be utilized to investigate the toxicological properties of other environmental contaminants. 相似文献
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Winter flounder (Pleuronectes americanus) from Boston Harbor, Massachusett, undergo a progressive series of hepatic changes involving hydropic vacuolation of epithelial cells, biliary hyperplasia, and hepatocellular and cholangiocellular neoplasia. Severely affected fish often exhibit grossly visible lesions. To examine cell proliferation associated with these conditions, evidence for DNA synthesis was sought. Boston winter flounder livers were screened endoscopically to select fish with and without grossly visible hepatic lesions, and then injected intraperitoneally with bromodeoxyuridine (BrdU) 3 h before euthanasia. Incorporation of this nucleotide analog was visualized immunohistochemically in formalin-fixed, paraffin-embedded tissues. Basal epithelia in the gill and intestine stained strongly, as did renal hemopoietic epithelia. In contrast, renal tubular epithelia and cardiomyocytes did not stain. Normal appearing hepatocytes stained occasionally, whilst more frequent staining was observed in vacuolated and neoplastic hepatic epithelia. We conclude (1) that the BrdU assay as described here has potentially widespread application to the study of cell proliferation in fish tissues; (2) that vacuolated cells may actively proliferate in winter flounder liver from Boston Harbor; and (3) vacuolated cells may be involved in the neoplastic transformation of hepatic epithelia. 相似文献
13.
Michael R. Miller David E. Hinton James J. Blair John J. Stegeman 《Marine environmental research》1988,24(1-4)
Monoclonal antibody directed against a major β-naphthoflavone (BNF)-induced form of teleost cytochrome P-450, P-450E (equivalent to P-450c in rat) was used to immunolocalize this enzyme in liver, gill and heart of scup and trout. Liver sections from both species showed P-450E in the cytoplasm of hepatocytes. No regional differences were observed which might indicate zonation of cytochrome P-450E within subpopulations of hepatocytes. Scup exocrine pancreatic cells were only weakly positive. In the gill of both fish, cytochrome P-450E was restricted to the endothelium (pillar cells) of secondary lamellae, where fluorescence appeared as a chain in longitudinal sections through lamellae and as star-shaped clusters in en face views. Sections of ventricular wall in both species revealed P-450E was restricted to endothelium at margins of muscle bands limiting heart ventricular lumen. Localization in the specific cells of these and other organs may be fundamentally important in understanding the role of cytochrome P-450E. 相似文献
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Harish C. Sikka A. Ruth Steward Sayed A. Elmarakby Subodh Kumar U. Devanaboyin Ramesh C. Gupta 《Marine environmental research》1995,39(1-4)
In order to examine factors that may contribute to the reported resistance of rainbow trout, Shasta strain, to the well-known hepatocarcinogenic effects of 2-acetylaminofluorene (AAF), the in-vitro and in-vivo metabolism of [14C]AAF in trout has been examined. Trout (compared to rat) liver microsomes metabolized AAF more efficiently, producing 3-fold larger amounts of ring-hydroxylated metabolites (7-hydroxy-AAF and 5-hydroxy-AAF), but 5-fold less N-hydroxy-AAF. Freshly isolated trout hepatocytes extensively metabolized AAF to form the same ring-hydroxylated metabolites and their respective glucuronide and sulfate conjugates. N-OH-AAF (plus its conjugates) and covalently-bound AAF derivatives amounted, respectively, to < 1% and 1.4–1.6% of total metabolites. Liver DNA of trout treated with AAF contained a single AAF-DNA adduct identified as N-(deoxyguanosin-8-yl)-2-aminofluorene (the major persistent AAF-DNA adduct found in rat liver). The level of this adduct (12 attomoles/μg DNA) was about 1000-fold lower than the level of AAF-DNA adduct previously reported in rat liver. The data show that trout liver, compared to rat liver, is considerably less efficient in metabolizing AAF to carcinogenic metabolites, and more efficient in forming nontoxic products, thus possibly explaining, in part, the resistance of trout to AAF-induced hepatocarcinogenesis. 相似文献
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I~ODWrIONExtraction of DNA from Porphyra po~nsis is very difficult due tO its high content ofpolysaccharides. TO avoid the interference Of POlysaccharides, HOng et al. (1992) used LiCI tosoften Porphyra thalli cells to release DNA directly without lysing the thalli, hoWever, the procedure is problematic in the isolation of high-quality DNA. Kitade et al. (1996) established a complicated procedure which includes the grindiflg of material in liquid nitrogen, lysing of material inextra… 相似文献
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条斑紫菜高纯度总DNA及其质粒状DNA的提取 总被引：11，自引：2，他引：9
提取条斑紫菜高纯度总DNA及其质粒状DNA的新方法。先用海螺酶处理紫菜叶状体制备细胞,然后用SDS-蛋白酶K裂解细胞提取总DNA,再用玻璃粉浆(glassmilk)对其纯化,经纯化后的总DNA能被EcoRI,Dral与HaeⅢ等限制酶完全酶切,并在酶切图谱上形成明显的DNA带型。当用异硫氰酸胍一十二烷基肌氨酸钠裂解紫菜细胞时,在总DNA提取物中直接发现有一条质粒状DNA带(2.3Kb),即建立了一种极简便的质粒状DNA提取方法。 相似文献
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G. Flouriot G. Monod Y. Valotaire A. Devaux J.-P. Cravedi 《Marine environmental research》1995,39(1-4)
Rainbow trout hepatocytes were isolated by a two step perfusion technique and maintained either in monolayer culture for 5 days or in aggregate culture for 30 days. Cytochrome P450 content decreased from day 0 to day 5 in both culture systems, and then was preserved at the same level after one month in aggregated cells. 7-Ethoxyresorufin O-deethylase (EROD) and UDP-glucuronosyl transferase were not significantly different in freshly isolated cells and in 30-day aggregated hepatocytes, whereas a substantial increase in glutathione S-transferase was observed. Two-day exposure of cells to β-naphthoflavone led to a significant increase in EROD activity in both culture systems, especially after one month of aggregation (10-fold increase). According to these results, aggregate culture of rainbow trout hepatocytes seems to be a promising in vitro model to investigate the biotransformation pathways in fish and their regulation by endogenous and exogenous compounds. 相似文献
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褐藻多糖硫酸酯对脂多糖诱导大鼠肾小球系膜细胞NO产生量的影响 总被引：2，自引：2，他引：0
慢性肾衰的进程中伴随着肾脏炎症的发生,研究表明褐藻多糖硫酸酯具有抗炎作用,且对于慢性肾衰的早期治疗具有一定的作用。本文通过脂多糖(lipopolysaccharide,LPS)诱导,建立大鼠肾小球系膜细胞的炎症模型,用来源于海带的高分子质量褐藻多糖硫酸酯F1和低分子质量褐藻多糖硫酸酯F2对细胞进行处理,24 h后MTT法检测各组细胞活力,Griess法检测24 h及48 h后细胞培养液中NO的含量。结果表明,各组间细胞活力无明显差异;脂多糖可以诱导大鼠肾小球系膜细胞NO产量增加,F1和F2均可在一定程度上抑制脂多糖诱导的NO产量的增加,从而减轻细胞炎症反应。剂量为25μg/m L的F1和F2处理细胞48 h后,与模型组相比,F1和F2组细胞培养液中NO含量分别下降22.52%和38.65%,低分子质量褐藻多糖硫酸酯抗炎效果明显高于高分子质量褐藻多糖硫酸酯。 相似文献
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采用四甲基偶氮唑盐(MTT)法体外肿瘤细胞株抑制试验，研究耳壳藻内酯合成过程中结构的衍化物和抗肿瘤活性的关系。结果表明．耳壳藻内酯对小鼠肝细胞癌(H22)、小鼠路易斯肺癌(LLC)、人宫颈癌(Hela)和人乳腺癌(MCF-7)有较强的细胞毒性，对正常小鼠胚成纤维(NIH3T3)和人肝细胞(L—02)的毒性较低。酮基内酯对各肿瘤细胞毒性差异较大，对上述正常细胞株的杀伤力也较大。体外结合DNA的研究表明，耳壳藻内酯芳香环上的羟基较酮基内酯上的酮基易于插入DNA分子中．造成DNA分子的断裂和解聚。标记代谢物前体掺入试验表明，耳壳藻内酯能够显著抑制DNA大分子的合成，阻滞肿瘤细胞的生长。推测耳壳藻内酯芳香环上的羟基是影响其抗肿瘤活性的主要官能团。 相似文献