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采用胶原酶二步灌流法获取怀孕大鼠的原代肝细胞,以Aroclor1254诱导肝细胞损伤,用不同剂量的槲皮素分别处理损伤的肝细胞24～72h,RT—PCR及Western—blot法检测肝细胞中细胞色素酶P450（CYP450）的表达。结果显示,槲皮素处理损伤的肝细胞后,肝细胞CYPIAl、CYP281及CYP2E1的表达随槲皮素浓度的增加和处理时间的延长而呈先升高后降低的趋势。10mg／LAroclor1254是诱导体外培养的原代肝细胞损伤的最适质量浓度,10μmol／L槲皮素是对损伤的怀孕大鼠肝细胞的最佳保护浓度。结果表明,槲皮素对损伤的怀孕大鼠肝细胞具有保护作用。 相似文献
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采用胶原酶二步灌流法获取怀孕大鼠的原代肝细胞,以Aroclor1254诱导肝细胞损伤,用不同剂量的槲皮素分别处理损伤的肝细胞2472h,RT-PCR及Western-blot法检测肝细胞中细胞色素酶P450(CYP450)的表达。结果显示,槲皮素处理损伤的肝细胞后,肝细胞CYP1A1、CYP2B1及CYP2E1的表达随槲皮素浓度的增加和处理时间的延长而呈先升高后降低的趋势。10mg/L Aroclor1254是诱导体外培养的原代肝细胞损伤的最适质量浓度,10μmol/L槲皮素是对损伤的怀孕大鼠肝细胞的最佳保护浓度。结果表明,槲皮素对损伤的怀孕大鼠肝细胞具有保护作用。 相似文献
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探讨MAPK通路在镉诱导大鼠肝细胞凋亡中的作用.采用两步灌流法获得大鼠肝细胞,经过24 h培养,用醋酸镉、醋酸镉与MAPK抑制剂(p38抑制剂SB202190、JNK抑制剂SP600125、ERK抑制剂U0126)共同处理肝细胞.用MTT法检测细胞存活率,倒置显微镜和荧光显微镜观察细胞形态和凋亡,免疫组织化学法检测p38蛋白表达.结果表明,镉可极显著提高肝细胞磷酸化p38的表达量(P＜0).01),而SB202190能极显著降低其表达(P＜0.01).SB202190可以显著或极显著提高镉处理组细胞的存活率(P＜0.05或P＜0.01),减少变形细胞和凋亡细胞数量,但SP600125和U0126作用相反.说明镉暴露导致肝细胞p38 MAPK途径激活而引起细胞凋亡. 相似文献
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用两步灌流法获得大鼠肝细胞,应用四甲基偶氮噻唑蓝(MTT)比色法、Hoechst 33258荧光染色法和流式细胞仪检测醋酸镉对肝细胞存活率和凋亡率的影响,以及N-乙酰半胱氨酸(NAC)的保护效应.结果表明,肝细胞暴露于浓度为2.5、5、10μmol/L的醋酸镉后,细胞相对存活率显著下降(P<0.01),凋亡率显著升高(P<0.05),且具有剂量-效应关系;2 mmol/L的NAC可使细胞相对存活率明显提高(P<0.01),凋亡率下降.结果提示,一定浓度醋酸镉可引起肝细胞凋亡,NAC具有一定的保护作用. 相似文献
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蛹虫草多糖对四氯化碳诱导大鼠原代肝细胞损伤的保护作用研究 总被引：2，自引：2，他引：0
蛹虫草是一种药用真菌。从人工培养的蛹虫草中提取蛹虫草多糖(CMPS),采用Ⅳ型胶原酶灌流法分离大鼠肝细胞进行原代培养,研究蛹虫草多糖对CCl4诱导大鼠原代肝细胞损伤的保护作用。结果表明:蛹虫草的子实体和基质都含有丰富的多糖,其中基质中的多糖含量最高,约为子实体的2～4倍;CMPS可明显降低由CCl4诱导的损伤肝细胞培养上清液中谷草转氨酶(AST)、谷丙转氨酶(ALT)的活性水平和丙二醛(MDA)的含量水平,显著提高由CCl4诱导的损伤肝细胞培养上清液中谷胱甘肽过氧化物酶(GSH-Px)活性和诱导损伤肝细胞的存活率。试验结果提示CMPS对大鼠原代培养肝细胞损伤有直接保护作用,该作用可能与其抗氧化作用有关。 相似文献
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本研究旨在建立一种简单有效的分离大鼠肝细胞和kupffer细胞的方法。大鼠体外肝脏灌流,采用percoll密度梯度分离法分离肝细胞与kupffer细胞,糖原染色鉴定肝细胞,墨汁吞噬实验鉴定kupffer细胞。结果表明,此方法分离的肝细胞和kupffer细胞活性都在95%以上,新分离的肝细胞鹅卵石样,贴壁能力不是特别强,24 h可分化出不同形态。Kupffer细胞具有较强的吞噬能力和贴壁能力,可看到墨汁被细胞吞噬。结论:本实验建立的分离培养方法较稳定,为开展肝细胞和kuffer细胞的相关研究提供方法依据,也为建立各种体外细胞分离培养方法提供借鉴。 相似文献
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为研究亚硒酸钠对游离脂肪酸 (free fatty acid,FFA) 致体外培养大鼠肝细胞凋亡和线粒体膜电位降低的保护作用,本试验通过胶原酶两步灌流法分离培养肝细胞,在细胞培养液中添加FFA和亚硒酸钠 (0.1 μmol/L)测定细胞的凋亡情况、细胞功能和线粒体膜电位。结果表明,随着培养液中FFA浓度的升高,细胞凋亡加重,肝细胞功能受到影响,线粒体膜电位降低且FFA的这种作用呈剂量依赖性;亚硒酸钠能抑制由FFA引起的肝细胞的上述变化。 相似文献
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本试验旨在为蛋鸡肝脏细胞分离和制备方法进行优化。以结扎肠系膜后静脉远心端,不结扎输卵管静脉和胰十二指肠静脉的成年鸡肝细胞培养灌流法为对照组;以本试验建立的同时结扎该3条静脉的灌流方法为处理组,进行蛋鸡肝细胞培养,分别记录肝脏灌流的成功率、肝细胞数量和细胞活率。同时,分别于试验第4、24、48、144、168小时测定肝细胞培养基上清液中乳酸脱氢酶(LDH)活力。结果显示,优化后的方法可使蛋鸡肝脏灌流的成功率提高至100%,肝细胞数量显著增加至(1.09±0.21)×109个(P<0.05),细胞活率也显著提升至(95.3±1.3)%(P<0.05)。两种方法培养的肝细胞培养基上清液LDH活力无显著差异(P>0.05)。本试验建立的肝细胞分离和制备方法比原有方法,更适用于产蛋家禽肝细胞的分离和制备。 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