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1.
试验选取540尾黄颡鱼(Pelteobagrus fulvidraco),对其连续投喂含不同水平香菇多糖(Lentinan,LNT)的配合饲料8周。采用Percoll密度梯度离心等技术,对黄颡鱼头肾巨噬细胞与外周血白细胞进行分离纯化,使用NBT还原法、Griesse试剂显色法与MTT法测定头肾巨噬细胞呼吸爆发活性与外周血白细胞的增殖能力,以探讨香菇多糖对黄颡鱼免疫细胞活性的影响。结果表明香菇多糖各水平组均能提高黄颡鱼头肾巨噬细胞的氧呼吸爆发活性,且1200~1500mg/kg水平组提高作用极显著(P<0.01);香菇多糖各水平组能提高黄颡鱼头肾巨噬细胞氮呼吸爆发活性,其中600 mg/kg水平组活性显著提高(P<0.05),900~1 500 mg/kg水平组活性提高效果极显著(P<0.01);投喂香菇多糖还能显著提高黄颡鱼外周血白细胞的增殖能力(P<0.05),且600~1500mg/kg水平组促进作用极显著(P<0.01)。可见,香菇多糖可以有效地提高黄颡鱼免疫细胞的活性。  相似文献   

2.
采用51%Percoll非连续密度梯度离心法分离团头鲂头肾吞噬细胞,在光镜和电镜下观察所分离细胞的显微、超微结构及吞噬作用;以二氢罗丹明(DHR123)为荧光探针,流式细胞仪检测细胞在佛波豆蔻酸乙酯(PMA)刺激下产生的呼吸爆发活动;光镜下观察壳聚糖对吞噬细胞吞噬功能的影响。结果表明:分离得到的细胞具有吞噬细胞(巨噬细胞与中性粒细胞)的形态学及功能特征;3种壳聚糖组(100μg/mL)均能提高吞噬细胞的呼吸爆发功能,在发荧光细胞比例及细胞发荧光强度上均极显著高于对照组。其中水溶性壳聚糖组的发荧光细胞比例显著高于其他试验组,而在细胞发荧光强度上,普通壳聚糖作用相对较高;同时壳寡糖能增强吞噬细胞的吞噬活性。  相似文献   

3.
采用皮质醇混饲投喂的方法.模拟自然条件下的慢性应激,研究皮质醇对黄颡鱼咸鱼吞噬细胞呼吸暴发和血清补体旁路溶血活性(ACH50)的影响。研究结果与结论如下。皮质醇混饲投喂可使血清皮质醇浓度持续显著升高,各周对照组血清皮质醇含量均无显著性差异,证明采用皮质醇混饲投喂的方法可模拟自然慢性应激情况下的血清皮质醇浓度持续显著升高。从第2周开始头肾吞噬细胞的呼吸暴发功能显著下降(P〈0.05),证明皮质醇对头肾吞噬细胞的杀菌能力产生显著抑制作用。10mg/kg剂量组,ACH50在投喂第2、第5、第6周显著低于对照组(P〈0.05),100mg/kg剂量组,ACH50从第2周开始一直都极显著低于同期对照组(P〈0.01),证明皮质醇显著抑制血清补体旁路溶血活性,且血清皮质醇浓度越高其抑制作用越强。  相似文献   

4.
黄颡鱼“裂头”病是近年来发生的黄颡鱼的新型病害,对广东顺德地区(龙眼、勒流、锦丰、杏坛等)黄颡鱼发病池塘进行采样分析,经流行病学调查、组织病理检测、病原分离等,对黄颡鱼“裂头”病的病因进行筛查。在所患病鱼脑部和浓汁中均分离出一种生长缓慢、透明的针尖状菌落。经鉴定为鲶爱德华氏菌。  相似文献   

5.
【目的】建立黄颡鱼仔鱼(Pelteboagrus fulvidraco)卵黄蛋白的ELISA检测方法,探讨黄颡鱼仔鱼体内卵黄蛋白含量的发生规律及仔鱼投喂生物饵料的最佳时间,为研究黄颡鱼仔鱼早期营养和开口饵料提供依据。【方法】以黄颡鱼仔鱼为试验材料,卵黄蛋白抗血清为抗体,纯化的卵黄蛋白为抗原,建立间接竞争酶联免疫吸附反应(ELISA)检测方法,测定黄颡鱼仔鱼卵黄蛋白在不同发育时期(破膜43,48,57,65,72,81,89,96,105,113,120和129h)的变化情况。【结果】黄颡鱼仔鱼卵黄蛋白ELISA检测中,抗血清的最佳稀释倍数为1∶20 000倍,包被抗原的最佳质量浓度为125ng/mL,最适检测质量浓度为20~2 000ng/mL,批内变异系数为6.529%,批间变异系数为6.873%。利用建立的ELISA方法检测发现,黄颡鱼卵黄蛋白含量随发育时间的延长而逐渐递减,至96h时缓慢下降直至消失。【结论】利用ELISA方法能相对准确地检测黄颡鱼仔鱼卵黄蛋白含量的变化,破膜96h是黄颡鱼仔鱼由内源营养阶段转向外源营养阶段的转折点,此时开始投喂适口的生物饵料更有利于黄颡鱼仔鱼开口摄食及生长发育。  相似文献   

6.
环境中污染物生物标志物的选取和研究是环境监测的重要内容之一,作为污染物肝脏解毒作用的关键酶,多功能氧化酶(CYP1A1)和谷胱甘肽硫转移酶(GST)常被用来作为多种污染物监测的生物标志物。比较了3个湖泊(巢湖、洪泽湖和鄱阳湖)黄颡鱼(Pelteobagrus fulvidraco)体内的CYP1A1和GST的活性,并对其进行功效分析。结果显示,3个湖泊中黄颡鱼体内CYP1A1和GST之间呈现反相关关系;洪泽湖黄颡鱼体内CYP1A1值明显小于其他两个湖,而洪泽湖黄颡鱼体内GST值明显大于巢湖,与鄱阳湖相比差  相似文献   

7.
为研究不同病原性相关分子模式(PAMP)对大黄鱼头肾巨噬细胞免疫功能的影响,本研究分别将脂多糖(lipopolysaccharide,LPS)、肽聚糖(peptidoglycan,PGN)、脂磷壁酸(lipoteichoic acid,LTA)、胞壁酰二肽(muramyl dipeptide,MDP)与提取的大黄鱼头肾巨噬细胞进行孵育,测定大黄鱼头肾巨噬细胞的呼吸爆发活性、超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性、溶菌酶(LZM)活性和免疫相关细胞炎症因子(TNFα,IL1β,IL8,IL10,CXCL9,MYD88,TLR3和TLR8)的表达情况。结果显示:(1)LPS,PGN,LTA,MDP均能显著提高大黄鱼头肾巨噬细胞的呼吸爆发活性和LZM活性,同时显著降低SOD和CAT两种抗氧化酶的活性;(2)在LPS和LTA刺激下,8种免疫相关细胞炎症因子TNFα,IL1β,IL8,IL10,CXCL9,MYD88,TLR3和TLR8均呈显著性上调趋势;在PGN和MDP刺激下,除IL8,CXCL9外,其余6种免疫相关细胞炎症因子均显著性上调。结果表明,4种病原相关分子模式LPS,PGN,LTA,MDP均能显著影响大黄鱼头肾巨噬细胞的非特异性免疫应答能力和免疫相关细胞炎症因子的表达。  相似文献   

8.
皮质醇对黄颡鱼腹腔炎性吞噬细胞的影响   总被引:3,自引:2,他引:1  
炎症是免疫系统对感染的重要应答.以皮质醇混饲投喂的方法,模拟自然发生的应急反应,研究了应激激素皮质醇对黄颡鱼腹腔渗出炎症细胞的影响.结果表明:通过腹腔注射液体石蜡后诱导的黄颡鱼腹腔渗出炎症细胞主要包括嗜中粒细胞、巨噬细胞和嗜酸性颗粒细胞(EGC).注射后2d,腹腔渗出细胞以嗜中性粒细胞为主(占嗜中粒细胞和巨噬细胞总数的61.5%±1.1%);注射后6d、10d则以巨噬细胞为主(在嗜中粒细胞、巨噬细胞和EGC分类计数中分别占77.8%±2.8%和82.3%±1.1%);注射后6d出现EGC,随后数量逐渐增加.炎症过程中腹腔渗出嗜中粒细胞和巨噬细胞对福尔马林灭活金黄色葡萄球菌(F-SA)的吞噬率和硝基四氮唑蓝(NBT)反应阳性细胞率逐渐上升.皮质醇混饲投喂使腹腔渗出巨噬细胞比例和吞噬细胞的吞噬率在注射液体石蜡后6d时显著下降,而EGC比例显著上升.  相似文献   

9.
黄颡鱼"裂头"病是近年来发生的黄颡鱼的新型病害,对广东顺德地区(龙眼、勒流、锦丰、杏坛等)黄颡鱼发病池塘进行采样分析,经流行病学调查、组织病理检测、病原分离等,对黄颡鱼"裂头"病的病因进行筛查.在所患病鱼脑部和浓汁中均分离出一种生长缓慢、透明的针尖状菌落.经鉴定为鲶爱德华氏菌.  相似文献   

10.
[目的]评估猪血多肽对草鱼的免疫增强作用,为进一步推广猪血多肽在水产养殖中的应用提供参考依据.[方法]采用密度梯度离心技术分离纯化草鱼头肾细胞、巨噬细胞和外周血白细胞,分别体外暴露在不同质量浓度的猪血多肽(0、10、20、40、80 μg/mL)中作用24 h,然后测定草鱼外周血白细胞增殖、巨噬细胞呼吸爆发活性和头肾细胞超氧化物歧化酶(SOD)活性,以探讨猪血多肽对草鱼体外培养免疫细胞活性的影响.[结果]猪血多肽体外作用24 h后,有效促进了草鱼体外培养免疫细胞的免疫活性,且以80 μg/mL猪血多肽的增效作用最明显,能极显著促进外周血白细胞增殖和增强巨噬细胞氧呼吸爆发活性(P<0.01),显著提高巨噬细胞氦呼吸爆发活性和头肾细胞SOD活性(P<0.05).[结论]猪血多肽对草鱼非特异性免疫具有促进作用,且作用效果呈剂量依赖性.实际生产中,可将猪血多肽开发成一种免疫添加剂,以增强水产动物对疾病的抵抗能力,提高水产养殖效益.  相似文献   

11.
Apoptotic cells expose phosphatidylserine and are swiftly engulfed by macrophages. Milk fat globule epidermal growth factor (EGF) factor 8 (MFG-E8) is a protein that binds to apoptotic cells by recognizing phosphatidylserine and that enhances the engulfment of apoptotic cells by macrophages. We report that tingible body macrophages in the germinal centers of the spleen and lymph nodes strongly express MFG-E8. Many apoptotic lymphocytes were found on the MFG-E8-/- tingible body macrophages, but they were not efficiently engulfed. The MFG-E8-/- mice developed splenomegaly, with the formation of numerous germinal centers, and suffered from glomerulonephritis as a result of autoantibody production. These data demonstrate that MFG-E8 has a critical role in removing apoptotic B cells in the germinal centers and that its failure can lead to autoimmune diseases.  相似文献   

12.
In this paper, toll-like receptor expression pattern in monocytes-derived macrophages by lipopolysaccharid (LPS) stimulation was examined. Jugular venous blood samples from 4 Japanese calves were obtained and the peripheral blood mononuclear cells (PBMC) were isolated. The PBMC were cultured for 7 d so as to collect monocytes-derived macrophages in Repcell. The PBMC were stimulated by LPS for 24 h and the mRNA expression pattern of TLR and cytokines in monocytes-derived macrophages (Mod-Mφ) was analyzed. Results showed that LPS stimulation of Mod-Mφ could increase the mRNA levels of the genes of TNF-α, IL-6, and IL-8. In addition, the mRNA levels of the genes of TNF-α and IL-6 in the group of LPS stimulation were most significantly (P 〈 0.01) higher than those in control group and the mRNA levels of TLR1, 3, 5, 8, and 10 were significantly (P 〈 0.05) decreased after LPS stimulation. There was no difference in the mRNA expressions of TLR2, 4, 6, and 7 between the groups of the control and LPS stimulation. Besides, expression of TLR9 was not found. It suggested that monocytes-derived macrophages could respond to LPS and they might take an important role in the innate immunity. The important function of the cells might contribute to better disease treatment.  相似文献   

13.
Pulmonary macrophages and plutonium particles were removed by washing the lungs of rats that had inhaled plutonium oxide-(239)Pu. A significant amount of plutonium was found in multiple washings of the same lung. The removal of toxic particles by washing is of potential therapeutic value. Particles were phagocytized by macrophages during the first 3 hours and retained within these cells for up to 25 days. Nearly all particles in washings were found in macrophages after the second day. The percent of macrophages with engulfed particles increased with increasing amounts of plutonium deposited in the lungs. The ability of pulmonary macrophages to rapidly phagocytize and retain plutonium particles deposited in the lungs has been shown.  相似文献   

14.
Macrophages have been generated from bone marrow progenitor cells (BMPCs) with macrophage colony-stimulating factor (M-CSF). Rapamycin (rapa) is an immunosuppressive drug, which could prevent renal graft rejection and inhibit tumor growth to yield antiproliferative activity in a variety of malignancies. However, the direct effect of rapa on the development of macrophages is unknown. In this study, we explored the direct effect of rapa on the differentiation and function of macrophages differentiated from mouse BMPCs in vitro in the presence of M-CSF. The experimental data showed that rapa prevented the differentiation of macrophagcs by down-regulating CD80 and CD86 expression but upregulating F4/80 expression, as well as by reducing the capacity of differentiated macrophages to stimulate lymphocyte proliferation in the allogeneic mixed lymphocyte reaction. Furthermore, the phagocytic capacity of differentiated macrophages was significantly reduced by rapa. Therefore, rapa may directly inhibit macrophage differentiation and function, which may have been one of the major targets for rapa to mediate its immunosuppressive properties.  相似文献   

15.
用睾酮诱导,研究小鼠骨髓巨噬细胞(BMMs)凋亡过程中Fas/FasL途径上caspase-8表达的改变。以BMMs经L929条件培养基(LCM)诱导5d后,用流式细胞仪分选出F4/80阳性细胞,并将细胞分为两大组。第1组是空白对照组,睾酮(100nmol·L-1,下同)处理组,去除LCM组,去除LCM同时用睾酮处理组。第2组用FADD反义寡核苷酸(ASODN)转染BMMs后,重复第1组的4个处理组,并以FADD错义寡核苷酸(MSODN)转染后的睾酮处理组作为对照。处理12h后,用流式细胞仪检测巨噬细胞的凋亡情况,并通过RT-PCR、Real-timeRT-PCR和WesternBlot方法检测各组中caspase-8基因及蛋白的表达。结果显示,在缺少LCM或用睾酮处理时,睾酮可在体外诱导小鼠骨髓巨噬细胞凋亡,并伴随caspase-8的活化。FADD反义寡核苷酸能抑制睾酮诱导的小鼠骨髓巨噬细胞的凋亡,其下游的caspase-8表达也被抑制。  相似文献   

16.
刺五加对巨噬细胞产生NO与肿瘤坏死因子-α的抑制作用   总被引:1,自引:0,他引:1  
用不同浓度刺五加提取物培养RAW264.7巨噬细胞,采用LPS与IFN-γ作为诱导剂,37℃培养24h,Griess试剂检测培养上清液中的NO含量,ELISA法检测培养上清液中的TNF-α含量。结果表明:刺五加提取物在浓度10~1000mg/L内对RAW264.7巨噬细胞分泌的NO具有极显著抑制作用(P<0.01),且呈剂量和时间依赖关系;刺五加提取物对TNF-α的分泌没有影响。这表明刺五加提取物可以显著抑制RAW264.7巨噬细胞产生的NO,而对肿瘤坏死因子-α的产生无明显抑制作用。  相似文献   

17.
[目的]研究强力天麻杜仲胶囊对小鼠巨噬细胞活性的影响。[方法]将强力天麻杜仲胶囊,连续给小鼠灌胃2周,计数小鼠腹腔巨噬细胞总数。采用ELISA法检测腹腔巨噬细胞TNF-α和IL-1β分泌水平;通过腹腔巨噬细胞吞噬鸡红细胞试验检测小鼠巨噬细胞的吞噬能力。[结果]结果表明,强力天麻杜仲胶囊能够轻度抑制小鼠腹腔巨噬细胞数量,明显降低腹腔巨噬细胞TNF-α和IL-1β的分泌水平和抑制小鼠腹腔巨噬细胞吞噬鸡红细胞的能力。[结论]强力天麻杜仲胶囊可降低小鼠巨噬细胞活性。  相似文献   

18.
The immune system recognizes invaders as foreign because they express determinants that are absent on host cells or because they lack "markers of self" that are normally present. Here we show that CD47 (integrin-associated protein) functions as a marker of self on murine red blood cells. Red blood cells that lacked CD47 were rapidly cleared from the bloodstream by splenic red pulp macrophages. CD47 on normal red blood cells prevented this elimination by binding to the inhibitory receptor signal regulatory protein alpha (SIRPalpha). Thus, macrophages may use a number of nonspecific activating receptors and rely on the presence or absence of CD47 to distinguish self from foreign. CD47-SIRPalpha may represent a potential pathway for the control of hemolytic anemia.  相似文献   

19.
LPS与ATP共同诱导巨噬细胞中NLRP3炎症小体的激活   总被引:1,自引:0,他引:1  
【目的】以脂多糖(LPS)为刺激源,探索小鼠巨噬细胞(RAW264.7)中NOD样受体家族pyrin结构域蛋白3(NLRP3)炎性小体激活的响应机制。【方法】试验分为对照组、LPS组、LPS与ATP共同刺激组。ELISA检测NLRP3源性白介素-1β(IL-1β)表达情况;RT-PCR检测NLRP3、Caspase-1、白介素-1β(IL-1β)的转录水平;ELISA检测LPS与NLRP3其他激动剂(MSU、CPPD、SiO2、Alum)共同作用后IL-1β的表达以及PI染色检测细胞焦亡。【结果】与对照组相比,LPS刺激组对IL-1β的表达无显著影响,LPS/ATP共同刺激组IL-1β的表达显著升高;LPS/ATP刺激组NLRP3、Caspase-1、IL-1βmRNA的表达显著升高且LPS/ATP刺激组发生明显的细胞焦亡。【结论】LPS与ATP共同作用能够显著提高NLRP3、Caspase-1、IL-1β的表达,说明LPS对NLRP3炎性小体的激活是LPS与ATP共同作用实现的。  相似文献   

20.
2种多糖对鲤鱼离体培养免疫细胞活性的影响   总被引:5,自引:0,他引:5  
采用Percoll密度离心等技术,对鲤鱼的头肾巨噬细胞和外周血白细胞进行分离纯化,并离体培养。体外暴露不同浓度的黄芪多糖(Astragalus polysaccharides,APS)和香菇多糖(Lentinan,LNT)后,分别采用MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)法测定它们对鲤鱼外周血白细胞增殖的影响;NBT(nitroblue tetrazolium)还原法和Griess试剂显色法测定对头肾巨噬细胞的呼吸暴发的影响。结果显示,香菇多糖能显著诱导巨噬细胞的氧暴发活性,黄芪多糖则没有显著的诱导作用;香菇多糖低浓度时对细胞的氮呼吸暴发活性无显著影响,黄芪多糖能显著诱导氮呼吸暴发活性,随着两者作用浓度的增加均表现为高浓度抑制作用;香菇多糖和黄芪多糖都能显著促进鲤鱼外周血淋巴细胞的增殖。结果表明黄芪多糖和香菇多糖对离体培养鲤鱼免疫细胞有明显活性作用,对鲤鱼非特异性免疫和特异性免疫具有促进作用,是有潜力的鱼用免疫增强剂。  相似文献   

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