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为研究建立大鼠脂肪肝体外模型的方法,试验采用原代肝细胞体外培养的方法培养大鼠肝细胞.对正常贴壁培养的肝细胞用1.0,2.0,3.0,4.0,5.0 mmol/L DL-乙硫氨酸处理,分别测定处理前及处理后24,48小时细胞培养基上清液中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AsT)的活性以及细胞裂解液中甘油三酯(TG)的含量.结果表明:贴壁培养的正常肝细胞用DL-乙硫氨酸处理后,培养基中ALT、AST的活性均有不同程度升高,其中5.0 mmol/,L处理组升高明显,与对照组相比差异极显著(P<0.01);4.0,5.0 mmol/L组的细胞裂解液中TG含量与对照组相比差异极显著(P<0.01).结果说明试验建立了大鼠脂肪肝药物损伤的体外模型,可以为在细胞水平筛选治疗脂肪肝的药物提供参考模型. 相似文献
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为研究肝细胞脂肪变性模型,采用体外培养的大鼠原代肝细胞作为试验材料。对正常贴壁培养的肝细胞用5.0mmol/L的DL-乙硫氨酸处理48 h,采用油红O染色,光学显微镜成像系统观察肝细胞的变化,并测定生化指标。结果表明,对照组细胞内少见橘红色脂滴,模型组细胞内可见许多大小不一的橘红色的脂滴存在,且部分细胞内脂滴连接成片,与对照组比较有极显著差异(P0.01)。模型组ALT,AST活性和TG含量升高,与对照组比较差异极显著(P0.01)。从而建立了大鼠脂肪肝脂肪变性体外病理模型,并为细胞水平筛选治疗脂肪肝药物提供参考模型。 相似文献
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为探讨丹参对脂肪变性奶牛肝细胞的作用机制,采用胶原酶两步灌流法分离培养奶牛肝细胞,不同浓度DL-乙硫氨酸处理,油红染色观察其脂滴数量,MTT法测定肝细胞的活性,生化法检测TG含量。用不同浓度的丹参处理变性肝细胞后,生化法检测ALT、AST、TG、SOD、MDA的含量或活性,ELISA法测定TNF-α的含量,Western blot法检测NF-κB和CYP450的蛋白表达水平。结果显示,筛选出2 mmol/L的DL-乙硫氨酸作用24 h作为体外造模的最佳条件。用丹参处理后,能降低ALT、AST、TG和MDA的活性或含量,抑制TNF-α的释放,增加SOD活性。随丹参浓度增加和处理时间的延长,CYP450的表达呈逐渐降低的趋势。24 h时,丹参对NF-κB的表达无明显影响;48 h时,随丹参浓度的增加,NF-κB的表达呈逐渐降低的趋势。这表明丹参对奶牛脂肪肝有一定的治疗作用,其作用机制与促进脂质代谢、抑制肿瘤细胞坏死因子、抗脂质过氧化及降低NF-κB和CYP450的表达密切相关。 相似文献
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《中国畜牧杂志》2016,(1)
本试验旨在研究烟曲霉毒素B1(FB1)对鸡肝脏细胞的毒性作用。将体外培养的鸡肝脏细胞分为6组,每组6个重复,1组为对照组,2~6组为FB_1组,分别在培养基质中添加FB_1,使FB_1终浓度为2.5、5、10、20、40μg/mL。染毒培养48 h,检测肝脏细胞存活率以及培养基质、肝细胞相关生化指标。结果表明:2.5~40μg/mL FB_1组肝脏细胞存活率显著低于对照组(P0.05),10~40μg/mL FB_1组基质中总抗氧化能力(T-AOC)、过氧化物酶(POD)活性显著低于对照组(P0.05),2.5~40μg/mL FB_1组基质中乳酸脱氢酶(LDH)活性显著高于对照组(P0.05),20μg/mL FB_1组基质中总胆固醇含量显著低于对照组(P0.05),5~40μg/mL FB_1组丙二醛(MDA)含量和谷丙转氨酶(ALT)活性显著高于对照组(P0.05),5~40μg/mL FB_1组超氧化物酶(SOD)活性显著低于对照组(P0.05),40μg/mL FB_1组谷草转氨酶(AST)活性显著高于其余各组(P0.05);AST、ALT活性以及MDA含量与FB_1浓度呈正相关(P0.05),LDH、SOD活性以及T-AOC与FB_1浓度呈负相关(P0.05)。由此可以看出,FB_1具有明显的肝细胞毒性,氧化损伤是FB_1致肝细胞毒性的重要致病机制。 相似文献
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为探讨葛根对奶牛脂肪肝细胞的作用机制,采用胶原酶二步灌流法分离病牛的肝细胞,并观察细胞形态。用不同浓度的葛根水提物处理肝细胞,生化法检测细胞上清液中丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）活性,并测定细胞裂解液中甘油三酯（TG）、胆固醇（TC）、丙二醛（MDA）、超氧化物歧化酶（SOD）的活性及含量。结果显示,50、100、500 μg/mL葛根水提物能极显著降低肝细胞中ALT、AST、TG、TC、MDA活性及含量,增加SOD活性（P < 0.01）。葛根水提物能显著改善肝细胞的相关生化指标,其机制可能与调节脂质代谢、抗脂质过氧化作用相关。 相似文献
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为研究黄芪乙醇粗提物对猫肾(Crandell-Rees Feline Kidney, CRFK)细胞的毒性,利用50%(体积分数)乙醇,超声(50℃,280 W)提取黄芪粉末,制得黄芪乙醇粗提物(简称“粗提物AME”);在初步分析粗提物AME的3类组成物质含量基础上,测定在不同质量浓度粗提物AME作用下的CRFK细胞活力、中性红吞噬能力及裂解液总蛋白质量浓度,并分析了粗提物AME对过氧化氢处理(氧化诱导)的细胞活力及总氧自由基水平的影响。研究结果显示：粗提物AME中多糖、总多酚和总蛋白质量分数分别为(53.07±1.73)%、(13.80±0.78)%和(0.13±0.01)%;在作用96 h内,高质量浓度粗提物AME(2 000μg/mL)使CRFK细胞活力降低,低质量浓度粗提物AME(50～1 000μg/mL)使CRFK细胞活力增强;粗提物AME添加质量浓度≥500μg/mL时,CRFK细胞中性红吞噬率降低,CRFK细胞裂解液总蛋白质量浓度也显著降低;50～2 000μg/mL的粗提物AME对由过氧化氢诱导的CRFK细胞活力下降具有显著抑制作用,且对胞内总氧自由基具有清除作用。 相似文献
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《中兽医医药杂志》2020,(5)
研究湿生扁蕾乙酸乙酯提取物(GEE)对乙醇诱导人正常肝细胞(L02)氧化损伤的保护作用及其机制。用400μmol/L乙醇诱导L02细胞建立体外细胞氧化损伤模型,加入GEE 25μg/mL、50μg/mL、100μg/mL作用24 h,采用MTT法检测细胞存活率;检测细胞培养上清液中谷丙转氨酶(ALT)、谷草转氨酶(AST)、细胞内超氧化物歧化酶(SOD)和丙二醛(MDA)含量的变化;DCFH-DA探针染色流式细胞仪检测细胞内ROS水平;DTNB-GR动力学循环法检测胞内还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)的含量变化。结果表明,400μg/mL乙醇诱导L02细胞24 h可建立肝细胞氧化损伤模型。GEE在25～100μg/mL对L02细胞无损伤。与模型组比较,GEE中剂量、高剂量组能显著提高L02细胞存活率(P0.001);降低细胞上清液中ALT、AST含量(P0.001);使细胞内SOD活力显著提高(P0.001);MDA含量明显降低(P0.001);同时使细胞内ROS水平降低(P0.01)、GSH/GSSG比值提高(P0.001)。提示GEE对乙醇诱导的L02细胞损伤具有保护作用,其机制可能与其减轻氧化应激反应有关。 相似文献
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为了探讨蔓越橘提取物对高脂大鼠氧化损伤的作用。试验选取50只体重120 g左右的SD雄性大鼠为试验动物,随机分成正常组、模型组和高、中、低3个试验组,每组10只。饲喂试验8周后,分别测定各组大鼠肝脏中丙二醛(MDA)含量与超氧化物歧化酶(SOD)活性、血清中甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)的含量与丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)和碱性磷酸酶(ALP)的活性。试验结果显示,与模型组比较,中、高剂量组中的TG降低显著(P<0.05),且试验组低剂量组的HDL-C量变化显著(P<0.05);饲喂蔓越橘提取物的试验组血清ALT、AST和ALP活性均有明显上升,并呈现一定剂量效应关系;试验各组大鼠肝脏中SOD活性明显增强(P<0.01),而MDA的生成量逐渐减少。结果表明,蔓越橘提取物能减轻高脂大鼠肝脏氧化损伤和脂质蓄积。 相似文献
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为了研究水栀子多糖提取物(Gardenia jasminoide var.radicans polysaccharide extract, GJPE)对猪圆环病毒2型(PCV-2)感染的RAW264.7细胞氧化应激的调节作用,试验采用ELISA法测定了GJPE对RAW264.7细胞的安全浓度;将RAW264.7细胞分为空白对照组、维生素C组、不同浓度(安全浓度范围)GJPE组,分别加入到96孔细胞培养板(1×106 个/mL)和24孔细胞培养板(2×105 个/mL)中,除空白对照组加基础培养基外,其他各组分别加入1×103 TCID50 PCV-2病毒液,并设置PCV-2组(只添加PCV-2病毒液)作为病毒对照组,100μL/孔(96孔细胞培养板)或500μL/孔(24孔细胞培养板),培养24 h;吸弃上清液,用PBS洗涤3遍,空白对照组和PCV-2组分别加入基础培养基,维生素C组加入200μmol/mL维生素C,不同浓度GJPE组分别加入相应浓度GJPE[100μL/孔(96孔细胞培养板)或50... 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
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本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
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以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
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REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献