首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到19条相似文献,搜索用时 125 毫秒
1.
通过对油酸诱导时间及有效作用浓度的研究,建立油酸致建鲤肝细胞脂肪变性模型。本研究以胰蛋白酶消化法获得的原代肝细胞作为实验材料,加入不同浓度油酸(0、0.05、0.1、0.2、0.4、0.8 mmol/L)与肝细胞共培养24–48 h,诱导肝细胞脂肪变性,分别收集不同时段的肝细胞及上清液,采用试剂盒测定肝细胞中甘油三酯(TG)、总胆固醇(TC)的含量,同时测定上清培养液中谷丙转氨酶(GPT)、谷草转氨酶(GOT)、γ-谷氨酰转肽酶(γ-GT)、乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)的活性,MTT法测定肝细胞的存活情况,确定油酸最佳诱导浓度,油红O染色法观察细胞内脂肪滴的形成情况。研究结果显示,0.4 mmol/L油酸与肝细胞共培养48 h,可以显著提高肝细胞内TG、TC含量(P﹤0.05或P﹤0.01),极显著提高上清培养液中GOT、GPT、LDH、γ-GT的活性,显著降低上清培养液中SOD的活性,但与24 h相比有升高趋势,0.4 mmol/L油酸与肝细胞共培养24 h,光镜下可见细胞内有脂肪滴形成,以48 h最明显。综上所述,油酸浓度0.4 mmol/L、诱导时间48 h成功构建了肝细胞脂肪变性模型,为研究鱼类脂肪肝类疾病奠定了基础。  相似文献   

2.
为了筛选草鱼肝细胞脂肪变性的最佳诱导剂及浓度,并初步分析脂肪乳剂(lipid emulsions,LE)引起草鱼肝细胞脂肪变性的作用机理,以草鱼(Ctenopharyngodon idellus)正常肝细胞为研究对象,建立草鱼脂肪变性肝细胞模型,以含10%胎牛血清的基础培养液为对照组,处理组为含20%脂肪乳剂0.5~2 m L/L和含20%、50%胎牛血清的诱导培养液,孵育草鱼肝细胞48 h后,定量分析肝细胞内的甘油三酯(TG)含量,观察脂滴积聚情况及肝细胞超微结构的变化,检测细胞培养上清中谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)的活性,q RT-PCR技术检测脂代谢关键基因(PPAR?、PPAR?、SREBP-1c、LPL、Lep和UCP2)的转录水平变化,蛋白质印迹技术检测PPAR?、SREBP-1c的蛋白水平变化。结果发现,含1~2 m L/L LE的诱导液组和含20%、50%FBS的诱导液组与对照组相比TG含量均显著上升(P0.05),且20%FBS和各浓度LE诱导组的转氨酶活性与对照组相比差异不显著(P0.05),表明含1~2 m L/L LE的诱导液和含20%FBS的诱导液均可建立草鱼营养性脂肪肝细胞模型。在肝细胞脂变模型中,PPARγ和LPL等脂代谢基因的表达量显著升高(P0.05),而Lep基因表达量显著降低(P0.05),PPARγ和SREBP-1c的蛋白水平升高。结论认为:采用1~2 m L/L LE和20%的FBS均可以在短时间内建立草鱼肝细胞脂肪变性模型,含1 m L/L LE的诱导液诱导效果最佳;肝细胞内脂质的蓄积可能与脂肪代谢关键基因PPARγ、SREBP-1c、LPL及Lep等密切相关。  相似文献   

3.
为了评价水飞蓟素(Silymarin, SM)对鲫(Cyprinus carpio)肝(细胞)的保护和抗氧化作用,分别从体外和体内两个方面探讨其作用效果.在体外,利用8 mmol/L的四氯化碳(CCl4)作用肝细胞4 h构建鲫肝细胞体外损伤模型,用不同质量浓度的水飞蓟素(100、300和600μg/mL)对鲫肝细胞进行前处理(CCl4损伤前加药)、后处理(CCl4损伤后加药)和前后处理(CCl4损伤前后均加药),然后检测细胞活力和细胞上清中谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量.在体内,用含水飞蓟素的饲料(0.1、0.5和1.0 g/kg)饲喂鲫60 d,再腹腔注射30%四氯化碳-植物油混合液,然后检测血清和肝组织匀浆中谷丙转氨酶(GPT)、谷草转氨酶(GOT)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)等生化指标.研究结果表明,水飞蓟素不同程度地抑制 CCl4损伤引起的肝细胞培养上清中 GPT、GOT、LDH活性和 MDA含量的升高,提高 SOD活性及肝细胞的存活率.体内研究发现,0.5和1.0 g/kg水飞蓟素显著降低CCl4损伤导致的血清中GPT、GOT、LDH活性升高,显著提高总蛋白(TP)、白蛋白(Alb)含量,有效提高肝中谷胱甘肽(GSH)含量、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)、总抗氧化能力(T-AOC)和超氧化物歧化酶(SOD)活性,降低丙二醛(MDA)含量(P<0.05或P<0.01);而0.1g/kg的水飞蓟素除了抑制LDH显著升高外,对其他指标均无效果.综合以上结果认为,水飞蓟素对鲫肝有保护作用,可防治鱼类肝(细胞)损伤,该保护作用可能与其抗氧化作用有关.  相似文献   

4.
以苏丹红Ⅲ为诱变剂,研究了其对泥鳅(Misgurnus anguillicandatus)的急性毒性效应,在此基础上研究了苏丹红Ⅲ在不同浓度和染毒时间对泥鳅红细胞微核率及其肝脏中谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性的影响.试验表明,苏丹红Ⅲ对泥鳅的24h半致死浓度为1.020 g/L,48 h半致死浓度为0.740 g/L,48 h安全浓度为0.117 g/L.染毒一段时间后,泥鳅红细胞的微核率与对照组比较均显著升高,达最高值后趋于平缓;随着苏丹红Ⅲ浓度的升高,泥鳅红细胞微核率与对照组相比出现极显著差异的时间相应提前.染毒后各组的谷草转氨酶和谷丙转氨酶活性呈下降趋势,达到峰值后趋于平缓;染毒时间越长,酶活下降的幅度越大.泥鳅红细胞的微核率及其肝脏中的GOT和GPT活性均与苏丹红Ⅲ有明显的剂量-时间效应,研究结果为进一步明确苏丹红Ⅲ的危害提供了试验数据.  相似文献   

5.
本研究运用细胞生物学研究方法,探讨中草药提取物黄连素对草鱼(Ctenopharyngodon idellus)肝细胞(L8824)活力、氧化应激和细胞凋亡的影响。用不同浓度黄连素(0μmol/L、5μmol/L、25μmol/L、50μmol/L和100μmol/L)分别处理体外培养的草鱼肝细胞Oh、6 h、12 h和24 h,采用CCK-8法检测细胞活力。结果显示,黄连素浓度和时间对细胞活力有显著交互作用(P0.05),且黄连素浓度对其活力影响差异显著(P0.05),草鱼肝细胞活力随着黄连素的浓度增加先升高后降低(P0.05)。在此基础上,设置了黄连素梯度浓度(0μmol/L、5μmol/L、25μmol/L、50μmol/L和100μmol/L)培养草鱼肝细胞12 h后,收集细胞,测定细胞谷丙转氨酶(GPT)及谷草转氨酶(GOT)活性,丙二醛(MDA)及活性氧(ROS)含量、细胞凋亡水平和相关基因mRNA表达量。结果表明,与不添加黄连素组相比,当黄连素浓度为100μmol/L时,细胞GOT和GPT活性、ROS和MDA含量显著(P0.05)增加。当黄连素的浓度为25μmol/L和50μmol/L时,细胞凋亡水平显著(P0.05)升高;当黄连素的浓度增加到100μmol/L时,细胞凋亡水平显著(P0.05)降低。当黄连素的浓度为50μmol/L和100μmol/L时,细胞半胱氨酸蛋白酶-3(caspase-3)mRNA表达量显著(P0.05)升高。综上所述,当黄连素浓度超过50μmol/L会抑制细胞的生长,引发氧化应激,影响细胞功能。因此,黄连素在草鱼肝细胞培养液中添加的适宜浓度为25~50μmol/L。  相似文献   

6.
谭娟  尚蕾  肖雅元 《淡水渔业》2011,41(3):39-42,21
用急性染毒法研究6种浓度(0、0.4、0.6、0.8、1.0、1.5 μg/L)的甲氰菊酯对(34.89±9.99)g的尼罗罗非鱼(Tilapia nilotica)分别处理6、12、24、48、96 h后,其组织内乙酰胆碱酯酶(AChE)、谷丙转氨酶(GPT)和谷胱甘肽( GSH)的动态变化.结果显示:AChE活性在...  相似文献   

7.
试验结果表明, 高效氯氰菊酯对鲫鱼的96 h LC50为11.4 μg/L.经0.114 μg/L、0.57 μg/L、1.14 μg/L质量浓度处理后,随着农药质量浓度的增高和暴露时间的延长,谷丙转氨酶(GPT)和谷草转氨酶(GOT)的活力升高,表明此农药对鲫鱼机体的损伤也增强.GPT和GOT活性可以作为杀虫剂对环境安全性的一种评价标准.  相似文献   

8.
用0、0.1、0.5、1.0 mg/L浓度的HgC l2对克氏原螯虾染毒后,进行血清生化指标检测。结果表明:总蛋白、球蛋白、白蛋白在较高浓度Hg2 作用下上升,而在低浓度Hg2 作用下总蛋白、球蛋白含量下降;甘油三酯和胆固醇含量随Hg2 的升高呈不同程度的下降趋势。谷草转氨酶(GOT)、谷丙转氨酶(GPT)、乳酸脱氢酶(LDH)、肌酸激酶(CTK)活性在高浓度Hg2 作用下呈应急性上升。  相似文献   

9.
采用胸腔注射法,研究了恩诺沙星对湘云鲫(Triploid crucian carp)组织谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性的影响。结果显示,给药后5 d,各试验组GPT、GOT酶活性均升高,与对照组之间呈显著差异水平(P<0.05);给药后10 d,各浓度组GPT酶活性有所降低,而GOT酶活性则继续升高,鳃的GOT酶活性达到了最大值;给药后15、20 d,各浓度组GPT酶活性继续下降,而肝脏、肾脏GOT酶活性在给药后15 d时达到最大值,之后开始下降;给药后25 d,鳃的GPT酶活性除200 mg/kg组外都恢复至正常水平,而肝脏、肾脏各浓度组的GPT酶活性显著低于对照组(P<0.05),肝脏的GOT酶活性恢复到了正常水平,而鳃和肾脏的GOT酶活性略高于对照组水平。  相似文献   

10.
以泥鳅(Misgurnus anguillicaudatus)为受试动物,采用急性毒性、遗传毒性及生理毒性试验,研究了[C8Mim]Cl对生物体的毒性效应.结果表明,[C8Mim] Cl对泥鳅有一定的毒性,其对泥鳅24h、48 h的LC50分别为324.13 mg/L、262.77mg/L,安全浓度为51.80 mg/L;各处理组的微核率均高于对照组(P<0.05),且随着处理浓度的升高和时间的推移,红细胞微核率都有明显的升高;处理组血清中的谷丙转氨酶(GPT)和谷草转氨酶(GOT)活力均明显高于对照组(P<0.05),且随离子液浓度升高以及时间的推移而增高;[C8Mim] Cl对泥鳅有显著的毒性作用.  相似文献   

11.
Fatty liver is an increasingly serious disease of fish in aquaculture. However, the mechanisms responsible for the occurrence of fatty liver remain unclear, and no effective methods for the prevention and treatment of this disease have yet been found. In the present study, we aimed to develop an in vitro model of hepatocyte injury using oleic acid as hepatotoxicant and evaluate the protective effects of Rhizoma Alismatis extract (RAE) in Jian carp using this model. Primary hepatocytes from Jian carp were isolated and purified and cultured in vitro. The result indicated that 0.4 mmol L?1 oleic acid and 48 h could be the optimal conditions to induce hepatocyte injury model in cultured hepatocytes. Hepatocytes were exposed to oleic acid, followed by the addition of RAE at 0, 1, 5, 10, 20, or 50 μg mL?1. The hepatocytes and supernatant were then analyzed. RAE suppressed oleic acid-induced elevations in aspartate aminotransferase, alanine aminotransferase, triglycerides, total cholesterol, lactate dehydrogenase, alkaline phosphatase, cholinesterase, malondialdehyde, γ-glutamyl transferase, cytochrome P450 1A, cytochrome P450 2E1, liver-type fatty acid binding protein, free fatty acid, fatty acid synthetase, and tumor necrosis factor-α (P < 0.01 or P < 0.05); reduced protein levels of cytochrome P450 1A, nuclear factor (NF)-κB p65, and NF-κB c-Rel; and inhibited cytochrome P4503A, NF-κB c-Rel, nuclear factor erythroid-related factor 2, peroxisome proliferator-activated receptor-α, and cytochrome P4501A mRNA levels. In conclusion, RAE exhibited a protective effect against hepatocyte injury in Jian carp. Further in vivo studies are needed to provide more evidence for the use of RAE as a hepatoprotective agent for the treatment of hepatocyte injury.  相似文献   

12.
This study investigated the effects of valine on growth, intestinal enzyme activities and microflora in juvenile Jian carp (Cyprinus carpio var. Jian). A total of 1200 fish with an average initial weight of 9.67 ± 0.03 g were fed diets containing 5.3 (unsupplemented control), 8.7, 11.8, 14.9, 18.7 and 20.1 g valine kg?1 diet for 60 days. Results indicated that the specific growth rate, feed efficiency, body protein and lipid content of fish were significantly improved by the dietary valine (< 0.05). The hepatopancreas weight and activities of trypsin, amylase, lipase, chymotrypsin, glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) took the similar trends. Similarly, the optimum levels of dietary valine induced increases in the intestinal length, weight, folds height and activities of alkaline phosphatase, gamma‐glutamyl transpeptidase and creatine kinase. In contrast, the trends of muscle GOT activity and plasma ammonia content were opposite. Intestinal Aeromonas, Escherichia coli, Lactobacillus and Bacillus were changed by dietary valine supplementations. The dietary valine requirement for Jian carp (9.67–76.4 g) based on SGR was 13.7 g valine kg?1 diet (4.0 g valine 100 g?1 CP). Together, these results indicated that valine improved fish growth, digestive and absorptive ability.  相似文献   

13.
A 9-week feeding trial was carried out with juvenile Jian carp to study the effect of dietary pantothenic acid (PA) on growth, body composition and intestinal enzyme activities. Semi-purified diets with seven levels (4.0, 15.5, 25.6, 36.1, 45.9, 56.1 and 65.9 mg PA kg−1) of supplemental calcium d -pantothenate were fed to Jian carp (13.0 ± 0.0 g). PA improved specific growth rate (SGR), protein productive value (PPV), protein efficiency ratio (PER) and lipid production value (LPV) ( P <  0.05). Fish fed the control diet had significantly lower feed efficiency (FE) than that in any other group ( P <  0.05). Body protein content increased with increasing PA levels ( P <  0.05), but moisture, lipid and ash of fish carcasses were negatively related to the graded PA levels ( P <  0.05). Intestine protein content (IPC), hepatopancreas protein content (HPC) and activity of α-amylase, lipase, trypsin, Na+,K+-ATPase, alkaline phosphatase (AKP) and gamma-glutamyl transpeptidase (γ-GT) were all positively affected by the dietary PA levels ( P <  0.05), while intestine index (ISI) and hepatopancreas index (HSI) decreased with the increment of supplemental levels of PA ( P <  0.05). These results suggested that PA could enhance fish growth and intestinal enzyme activities. The dietary PA requirement of juvenile Jian carp, Cyprinus carpio var . Jian (13.0–73.0 g), for optimal growth estimated by the broken-line analysis was 23.0 mg PA kg−1 diet.  相似文献   

14.
This study was designed to determine the effects of diet supplementation with uncoated lysine (UCL) or coated lysine (CL) to juvenile Jian carp Cyprinus carpio Var. Jian. 330 juvenile Jian carps (initial weight 7.89 ± 0.04 g) were randomly allocated to two groups, feeding a diet containing UCL or a CL for 80 days. Weigh gain, feed intake and feed efficiency ratio were higher in group CL than group UCL, indicating the effect of CL to be better than UCL. Hepatopancreas weight and protein content, intestinal weight, length and protein content, intestinal folds height, activity of intestinal protease, lipase, alkaline phosphatase (AKP), Na+, K+‐ATPase were positively affected by CL. Moreover, glutamate‐oxaloacetate transaminase (GOT) in muscle, and plasma ammonia, tended to be lower for CL, which means that the improvement of digestive organ development and function may be the possible reason why CL was better than UCL.  相似文献   

15.
This study was to investigate the effect of dietary pantothenic acid (PA) on the disease resistance, immune response and intestinal microflora on juvenile Jian carp (Cyprinus carpio var. Jian). Seven diets (4.0, 15.5, 25.6, 36.1, 45.9, 56.1 and 65.9 mg PA kg?1) were fed to Jian carp (12.95 ± 0.03 g) for 9 weeks. After 9‐week feeding trial, the challenge experiment with Aeromonas hydrophila was conducted to determine the impact of PA on fish disease resistance. Survival rate after challenge was promoted with the increasing PA levels (P < 0.05). Blood counts also significantly increased up to the dietary PA level of 25.6 mg PA kg?1 (P < 0.05). Leucocyte phagocytic activity, lectin potency, lysozyme and acid phosphatase activity, and total iron‐binding capacity were improved with increasing PA levels (P < 0.05). Serum immunoglobulin M level and agglutination antibody titre to A. hydrophila were increased (P < 0.05) in fish fed the diets with the dietary PA levels between 56.1 and 65.9 mg kg?1. PA also promoted the growth and reproduction of Lactobacillus and depressed Escherichia coli and A. hydrophila (P < 0.05). These results suggested that pantothenic acid could improve disease resistance, immune response, and the balance of intestinal microflora in juvenile Jian carp.  相似文献   

16.
Oxidative damage and antioxidant status of intestine and hepatopancreas for juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of methionine hydroxy analogue (MHA: 0, 5.1, 7.6, 10.2, 12.7, 15.3 g kg?1 diet) for 60 days were studied. Radical scavenging ability, antioxidant enzymes activities such as superoxide dismutase (SOD), catalase (CAT), glutathione‐S‐transferase (GST), glutathione peroxidase (GPX) and glutathione reducase (GR), as well as glutathione (GSH), protein carbonyl (PC) and malondialdehyde (MDA) contents were assayed in these tissues. Results indicated that anti‐superoxide anion capacity in intestine and anti‐hydroxyl radical capacity in hepatopancreas significantly improved with dietary MHA levels up to 7.6 and 10.2 g kg?1 diet respectively, whereupon they decreased (P < 0.05). SOD, CAT, GST, GPX, GR activities in intestine and hepatopancreas, as well as GSH content in hepatopancreas significantly increased with optimal MHA levels which were in the range of 5.1–10.2 g kg?1 diet, and thereafter decreased (P < 0.05). Meanwhile, MDA and PC contents in these tissues together with GOT and GPT activities in plasma significantly decreased with optimal MHA levels which were in the range of 5.1–7.6 g kg?1 diet, and thereafter increased (P < 0.05). These results suggested that MHA improved antioxidant status and depressed lipid and protein oxidation in intestine and hepatopancreas.  相似文献   

17.
建鲤苗种体长与体重间相互关系的分析   总被引:2,自引:0,他引:2  
盛东峰  郭翠红 《河北渔业》2009,(7):6-7,32,63
利用建鲤实测体长和体重数据,应用统计分析方法进行体长-体重线性回归分析.结果表明:建鲤体长和体重之间呈曲线相关关系,其相关公式为:W=0.011L3.077(R=0.9919)。  相似文献   

18.
In the present study, we tested the hypothesis that dietary histidine could improve antioxidant capacity of juvenile Jian carp (Cyprinus carpio var. Jian). A total of 1,200 juvenile Jian carp were fed graded levels of histidine at 2.3 (unsupplemented control), 4.4, 6.3, 8.6, 10.8 and 12.7 g/kg diet for 60 days. Results showed that the content of malondialdehyde (MDA) and protein carbonyl (PC) in serum and all tissues apparently decreased with increasing histidine levels up to an optimal level and increased thereafter. Anti-superoxide anion (ASA) capacity, glutathione peroxidase (GPX) activities and glutathione (GSH) content in serum and all tissues, anti-hydroxyl radical (a-HR) capacity, catalase (CAT) and glutathione-S-transferase (GST) activities in serum, muscle and intestine, superoxide dismutase (SOD) activities in serum and intestine, as well as glutathione reductase (GR) activity in serum, muscle and hepatopancreas were improved by dietary histidine. Fish fed diet with 8.6 g/kg histidine had lower serum glutamate-pyruvate transaminase (GPT) activity than that fed with control diet, whereas pattern of glutamate–oxaloacetate transaminase (GOT) activity was opposite. The present results suggested that histidine could improve antioxidant capacity and inhibit lipid peroxidation and protein oxidation of juvenile Jian carp.  相似文献   

19.
The present study aimed to evaluate the hepatoprotective effects of Angelica sinensis extract (ASE) against carbon tetrachloride (CCl4)‐induced hepatotoxicity in Jian carp (Cyprinus carpio var.Jian). Fish were fed diets containing four doses of ASE (0%, 0.1%, 0.5% and 1.0%) for 60 days, and then given an intraperitoneal injection of 30% CCl4 in olive oil at a volume of 0.05 mL/10 g body weight. At 72 h post injection, blood and liver samples were collected for biochemical analysis, comet assay, histopathological examination and CYP3A mRNA expression. Results showed that the increases of glutamate pyruvate transaminase (GPT) and glutamate oxalate transaminase (GOT) induced by CCl4 were significantly inhibited by pre‐treating the fish with 0.1%, 0.5% and 1.0% ASE in the diets. The elevation of lactate dehydrogenase (LDH) and the reductions of the total protein (TP) and albumin (Alb) in the serum induced by CCl4 were also inhibited by pre‐treatments with 0.5 and 1.0% ASE. In the liver tissue, pre‐treatment with 1.0% ASE significantly inhibited the malondialdehyde (MDA) formation and the reductions of the total antioxidant capacity (T‐AOC), superoxide dismutase (SOD), glutathione (GSH) and CYP3A mRNA expression induced by CCl4. Comet assay showed that tail moment, olive tail moment, tail length and tail DNA% were positively changed in fish pretreated with 0.5 and 1.0% ASE. CCl4‐induced histological changes were obviously reduced by 0.5% and 1.0% ASE. Overall results prove the hepatoprotective effect of ASE in a dose‐dependent manner and support the use of ASE (1.0%) as a hepatoprotective and antioxidant agent in fish.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号