| 摘要: |
| 作为典型的降河洄游鱼类,花鳗鲡(Anguilla marmorata)在我国主要分布于长江下游及以南水域。中草药作为抗生素等化学类渔药的绿色替代品,对养殖鱼类具有抗菌、消炎、促生长等多种功效。为研究中草药复方在花鳗鲡病害防治中的作用,扩大水产养殖业渔药研制与应用的选择,采用非连续性密度梯度离心法对花鳗鲡外周血白细胞的分离体系进行了优化,试验设置了密度分别为1.031、1.043、1.056、1.067、1.090 g/mL的五种分离液,分别采用染色观察法和流式细胞术检测各分离液中白细胞浓度;以浓度1.0mg/mL的五种复方药液分别与105cell/mL的白细胞悬液共孵育2h后,分别采用NBT法和流式细胞术测定白细胞的呼吸暴发活性和吞噬能力,评价五种中草药复方(多糖复方Dt,白头翁复方Btw,板蓝根复方Blg,大黄复方Dh,百里酚复方Blf)对花鳗鲡外周血白细胞的激活作用。结果显示:1.043g/mL和1.056g/mL的分离液所分离获得白细胞浓度均接近105cell/mL,可在后续实验中叠加使用该两个浓度的分离液分离获得白细胞;流式细胞术检测白细胞浓度比染色观察法更为灵敏准确;5种复方均能显著提高花鳗鲡外周血白细胞的呼吸暴发活性(P<0.05),大小依次为Blg>Dt>Btw>Blf>Dh;Dt和Blg能显著提高白细胞吞噬率(P<0.05),Btw、Blf、Dh也能提高白细胞吞噬率,但差异不显著(P>0.05);5种复方对发生吞噬白细胞比率的影响差异不显著(P>0.05)。 |
| 关键词: 白细胞呼吸暴发活性 白细胞吞噬能力 流式细胞术 中草药复方 花鳗鲡 |
| DOI：10.11693/hyhz20220300063 |
| 分类号:S942;S948;S965 |
| 基金项目:福建省科技计划农业引导性(重点)项目,2021N01010292号;福建省海洋渔业结构调整专项,闽财指[2021]66号;国家重点研发计划,2020YFD0900102号;财政部和农业农村部:国家现代农业产业技术体系资助,CARS-46号;福建省海洋经济发展补助资金项目,FJHJF-L-2020-4号;集美大学培育基金,ZP2021003号。 |
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| EVALUATION ON THE ACTIVITY OF FIVE CHINESE HERBAL COMPOUNDS IN ACTIVATING PERIPHERAL BLOOD LEUKOCYTES OF ANGUILLA MARMORATA IN VITRO |
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LI Zhong-Qin1,2, ZHANG Xin-Yan3, YANG Qiu-Hua4,2, LIN Mao1,2, JIANG Xing-Long1,2, ZHAI Shao-Wei1,2
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1.Fisheries College, Jimei University, Xiamen 361021, China;2.Engineering Research Center of the Modern Technology for Eel Industry, Ministry of Education, Xiamen 361021, China;3.Freshwater Fisheries Research Institute of Fujian, Fuzhou 350002, China;4.Fujian Fisheries Research Institute, Xiamen 361012, China
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| Abstract: |
| As a green substitute for chemical fishery drugs such as antibiotics, Chinese herbal medicines have many effects such as antibacterial, anti-inflammatory, and promoting the growth of cultured fish. To study the role of Chinese herbal compounds in the prevention and control of diseases to Anguilla marmorata and expand the selection of fishery medicine development and application in aquaculture, the separation system of leukocytes from peripheral blood of A. marmorata was optimized by discontinuous density gradient centrifugation. Five separation solutions in densities of 1.031, 1.043, 1.056, 1.067, and 1.090 g/mL were set up. The concentration of leukocytes in each separation solution was detected by staining observation and flow cytometry. After the five compound solutions in concentration of 1.0 mg/mL were incubated with 105 cell/mL leukocytes for 2 hours, the respiratory burst activity and phagocytic activity of leukocytes were analyzed by NBT method and flow cytometry, and the effects of the five Chinese herbal compounds on the activation of leukocytes in peripheral blood of A. marmorata were evaluated. Results show that the leukocyte concentration of 105 cell/mL was achieved from the separation solutions of 1.043 g/mL and 1.056 g/mL. Flow cytometry was more sensitive and accurate than staining method in detecting the concentration of leukocytes. The five compounds could significantly improve the respiratory burst activity of leukocytes from peripheral blood of A. marmorata (P<0.05), among them Radix isatidis (Banlangen, Blg) compound could maximally increase the leukocyte respiratory burst activity (P<0.05). These compounds could also increase the leukocyte phagocytosis rate to certain degrees, among them polysaccharide (Doutang, Dt) compound and Radix isatidis (Banlangen, Blg) compound could also significantly increase leukocyte phagocytic activity (P<0.05), but the three others among the five compounds was not significant (P>0.05). |
| Key words: leukocyte respiratory burst activity leukocyte phagocytic activity flow cytometry Chinese herbal compounds Anguilla marmorata |
