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2025年4月3日 星期四
农业生物技术学报  2022, Vol. 30 Issue (8): 1580-1593    DOI: 10.3969/j.issn.1674-7968.2022.08.013
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
复方中草药对虹鳟脾脏非特异性免疫指标及免疫相关基因表达的影响
潘玉财1, 黄进强1,*, 李永娟2, 吴深基1, 赵璐1, 雷明荃1, 孙同振1, 王晓谰1
1 甘肃农业大学 动物科学技术学院,兰州 730070;
2 甘肃农业大学 理学院,兰州 730070
Effects of Compound Chinese Herbal Medicine on Non-specific Immunity Parameters and Immune-related Gene Expressions in Spleen of Rainbow Trout (Oncorhynchus mykiss)
PAN Yu-Cai1, HUANG Jin-Qiang1,*, LI Yong-Juan2, WU Shen-Ji1, ZHAO Lu1, LEI Ming-Quan1, SUN Tong-Zhen1, WANG Xiao-Lan1
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2 College of Science, Gansu Agricultural University, Lanzhou 730070, China
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摘要 中草药具有丰富的活性物质,具有提高机体免疫力、抗病及无残留等特性,已在水产养殖中被广泛应用。为探讨饲料中添加不同水平的复方中草药(黄芪(Astragalus radix), 党参(Codonopsis pilosula), 当归(Angelica sinensis), 甘草(Glycyrrhiza uralensis), 麦冬(Ophiopogon japonicus), 茯苓(Poria cocos), 山楂(Hawthorn), 金银花(Lonicera japonica), 板蓝根(Isatidis radix)和大青叶(Isatis indigotica Fortune))对虹鳟(Oncorhynchus mykiss)脾脏非特异性免疫及免疫相关基因表达的影响,本研究选取120尾虹鳟为实验对象,随机分为4组,分别在基础饲料中添加0 (对照组)、10、20和30 g/kg复方中草药连续投喂35 d。在投喂第7、21和35 d时取脾脏组织用于非特异性免疫指标的测定,并利用qPCR分析虹鳟脾脏中15个免疫相关基因的表达差异。结果显示,与对照组相比,在投喂的3个时期复方中草药均能显著提高虹鳟脾脏中总超氧化物歧化酶(total superoxide dismutase, T-SOD)、过氧化氢酶(catalase, CAT)、溶菌酶(lysozyme, LZM)及酸性磷酸酶(acid phosphatase, ACP)的活性(P<0.05),其中以20 g/kg组效果最佳;与对照组相比,10、20和30 g/kg组丙二醛(malondialdehyde, MDA)含量均在投喂的3个时期显著下降(P<0.05),且30 g/kg组效果最佳。基因表达结果显示,与对照组相比,投喂第35 d时,20 g/kg组能显著诱导脾脏中Toll样受体3 (Toll-like receptor 3, tlr3)、Toll样受体7 (tlr7)、髓样分化因子88 (medullary differentiation factor 88, myd88)、白细胞介素1β (interleukin-1β, il-)和细胞核因子κB抑制剂α (nuclear factor kappa B inhibitor alpha, nfkbia)基因的表达(P<0.05);各处理组肿瘤坏死因子α (tumor necrosis factor-α, tnf-α)基因表达差异不显著,Toll样受体8 (tlr8)在30 g/kg组显著上调(P<0.05);同时,免疫相关基因干扰素调节因子3 (interferon regulatory factor 3, irf3)、干扰素调节因子7 (irf7)、黑色素瘤分化相关蛋白5 (melanoma differentiation-associated protein 5, mda5)、RIG-I样受体2 (RIG-I-like receptor 2, lgp2)、白细胞介素8 (il-8)、干扰素β (interferon-β, ifn-β)、信号转导和转录激活因子1 (signal transducer and activator of transcription 1, stat1)和janus激酶1 (janus kinase 1, jak1)表达量也在20 g/kg组中显著上调(P<0.05)。本研究表明,该复方中草药可以增强虹鳟非特异性免疫能力和免疫相关基因的表达,且饲料中复方中草药建议添加量为20 g/kg。本研究为促进复方中草药作为免疫刺激剂在水产养殖中的合理应用及发展提供了科学依据。
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潘玉财
黄进强
李永娟
吴深基
赵璐
雷明荃
孙同振
王晓谰
关键词 虹鳟复方中草药非特异性免疫免疫相关基因表达    
Abstract:Traditional Chinese herbs are rich in active ingredients and have the properties of improving the immunity of the body, disease resistance and residue-free, and have been widely used in aquaculture. The aim of this study was to investigate the effects of compound Chinese herbs medicine (composed of Astragalus radix, Codonopsis pilosula, Angelica sinensis, Glycyrrhiza uralensis, Ophiopogon japonicas, Poria cocos, Lonicera japonica, Isatidis radix, Isatis indigotica Fortune, Hawthorn) on the non-specific immunity parameters and immune-related gene expressions of rainbow trout (Oncorhynchus mykiss) and provide scientific basis for the healthy culture. The 120 healthy rainbow trouts with an average weight (33.0 ± 2.8) g were selected and randomly assigned into 4 groups; each group repeated the experiment 3 times and 10 rainbow trout per replicate. The fish were fed a basal diet supplemented with compound Chinese herbal medicine at concentrations of 0 (control group), 10, 20 and 30 g/kg, respectively. The feeding trial lasted 35 days. Subsequently, spleen tissue (4 per group) was collected at 7, 21 and 35 d of feeding to evaluate non-specific immune parameters. The expression of fifteen immune-related genes in spleen tissue was detected by qPCR after 35 d of feeding. The results showed that dietary compound Chinese herbal medicine significantly increased the levels of total superoxide dismutase (T-SOD), catalase (CAT), acid phosphatase (ACP) and lysozyme (LZM) in the rainbow trout spleen on the 7th, 21th and 35th day (P<0.05), and the 20 g/kg supplemental groups showed the best results. Furthermore, the malondialdehyde (MDA) concentration in the spleen was significantly lower in the compound Chinese herbal medicine treatment groups than that in the control group (P<0.05), and the 30 g/kg supplemental group showed the best results. Moreover, the gene expression data indicate that dietary compound Chinese herbal medicine significantly upregulated the expression of Toll-like receptor 3 (tlr3), Toll-like receptor 7 (tlr7), medullary differentiation factor 88 (myd88), interleukin-1β (il-1β), nuclear factor kappa B inhibitor alpha (nfkbia) in the 20 g/kg supplemental group (P<0.05). The tumor necrosis factor-α (tnf-α) gene expression in the treatment group was no difference compared to the control group and Toll-like receptor 8 (tlr8) gene was significantly upregulated in the 30 g/kg supplemental group (P<0.05). Additionally, immune-related genes included melanoma differentiation-associated protein 5 (mda5), RIG-I-like receptor 2 (lgp2), interferon regulatory factor 3 (irf3), interferon regulatory factor 7 (irf7), interleukin-8 (il-8), interferon-β (ifn-β), janus kinase 1 (jak1) and signal transducer and activator of transcription 1 (stat1) in the 20 g/kg supplemental group were significantly upregulated (P<0.05). In conclusion, the current results indicated that a diet supplemented with compound Chinese herbal medicine has positive effects on non-specific immune responses of rainbow trout, it suggested that the best adding level was 20 g/kg. This study provides a scientific basis for promoting the rational application and development of compound Chinese herbal immunostimulants in aquaculture.
Key wordsRainbow trout    Compound Chinese herbal medicine    Non-specific immunity    Immune-related gene expression
收稿日期: 2021-10-20     
ZTFLH:  S917.4  
基金资助:甘肃农业大学科技创新基金-青年导师扶持基金(GAU-QDFC-2018-09); 甘肃省农牧厅特种养殖专项基金(GARS-TSYZ-1-2); 甘肃省现代丝路寒旱农业科技支撑计划(GSLK-2021-16)
通讯作者: *huangjq@gsau.edu.cn   
引用本文:   
潘玉财, 黄进强, 李永娟, 吴深基, 赵璐, 雷明荃, 孙同振, 王晓谰. 复方中草药对虹鳟脾脏非特异性免疫指标及免疫相关基因表达的影响[J]. 农业生物技术学报, 2022, 30(8): 1580-1593.
PAN Yu-Cai, HUANG Jin-Qiang, LI Yong-Juan, WU Shen-Ji, ZHAO Lu, LEI Ming-Quan, SUN Tong-Zhen, WANG Xiao-Lan. Effects of Compound Chinese Herbal Medicine on Non-specific Immunity Parameters and Immune-related Gene Expressions in Spleen of Rainbow Trout (Oncorhynchus mykiss). 农业生物技术学报, 2022, 30(8): 1580-1593.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2022.08.013     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2022/V30/I8/1580
 
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