瓦氏黄颡鱼头肾组织学观察与先天性免疫屏障研究

陈杰; 贺扬; 戴旭平; 王均; 覃川杰; 李锐

doi:10.12131/20200119

瓦氏黄颡鱼头肾组织学观察与先天性免疫屏障研究

doi: 10.12131/20200119

陈杰^1,,
贺扬^{1, 2, ,},
戴旭平¹,
王均^{1, 2},
覃川杰^{1, 2},
李锐¹

1.
内江师范学院生命科学学院，四川内江 641000
2.
内江师范学院长江上游鱼类资源保护与利用四川省重点实验室, 四川内江 641000

基金项目: 四川省科技计划应用基础研究项目 (2018JY0286)；内江师范学院大学生创新创业孵化项目 (X2019033)

详细信息

作者简介:
陈杰：陈　杰 (1997—)，女，本科生，研究方向为水生生物疾病与病理。E-mail: 850301489@qq.com

通讯作者:
贺　扬 (1987—)，女，博士，副教授，从事水生生物疾病与病理研究。E-mail: 10001431@njtc.edu.cn

中图分类号: S 942.1
计量
- 文章访问数: 88
- HTML全文浏览量: 31
- PDF下载量: 35
- 被引次数: 0
出版历程
- 收稿日期: 2020-05-30
- 修回日期: 2020-09-07
- 网络出版日期: 2020-12-16
- 刊出日期: 2021-02-04

Histological observation and innate immune barrier study of head kidney of Pelteobagrus vachelli

Jie CHEN^1
,,
Yang HE^{1, 2
, ,},
Xuping DAI¹,
Jun WANG^{1, 2},
Chuanjie QING^{1, 2},
Rui LI¹

1.
College of Life Science, Neijiang Normal University, Neijiang 641000, China
2.
Sichuan Key Laboratory of Fish Resources Protection and Utilization in the Upper Reaches of the Yangtze River, Neijiang Normal University, Neijiang 641000, China

摘要

摘要: 为探明瓦氏黄颡鱼 (Pelteobagrus vachelli) 头肾组织结构和先天性免疫屏障，该研究用苏木素-伊红 (HE) 染色法和改良James染色法对瓦氏黄颡鱼头肾的显微结构进行了观察，并使用台盼蓝活体染色法对头肾免疫屏障部位进行了定位。结果显示，瓦氏黄颡鱼头肾由被膜、淋巴细胞聚集区、粒细胞聚集区、前肾间组织、网状纤维和胶原纤维组成，胶原纤维仅在血管外周可见；台盼蓝主要分布于血窦周围的内皮细胞和一些游离的巨噬细胞中，且随时间延长呈递增趋势。研究表明血窦周围的内皮细胞和巨噬细胞组成了瓦氏黄颡鱼的头肾先天性免疫屏障。
- 瓦氏黄颡鱼 /
- 头肾 /
- 组织结构 /
- 免疫功能
Abstract: To investigate the histological structure and immune function of head kidney of Pelteobagrus vachelli, we used hematoxylin-eosin (HE) staining and modified James staining to observe its microscopic structure, and applied trypan blue vital staining to locate its immune barrier site. The results show that the head kidney of P. wallichii consisted of capsule, lymphoid zone, granulocytic zone, anterior interrenal tissue, reticular fibers and collagen fibers, and collagen fibers were only visible at the periphery of blood vessels. Trypan blue was mainly distributed in endothelial cells around sinusoids and some free melanin macrophages, showing an increasing trend with time. Results suggest that endothelial cells of red pulps and free macrophages form the innate immune barrier of head kidney.
- Pelteobagrus vachelli /
- Head kidney /
- Tissue structure /
- Immune function

HTML全文

图 1 瓦氏黄颡鱼头肾解剖示意图

a. 头肾的解剖位置；b. 头肾的形态

Figure 1. Anatomical diagram of head kidney of P. vachelli

a. Anatomical location of head kidney; b. Anatomical structure of head kidney

下载: 全尺寸图片幻灯片

图 2 瓦氏黄颡鱼头肾的组织结构HE染色观察

a. 被膜 (→)，血窦 (▲)；b. 淋巴细胞聚集区 (○) 与粒细胞聚集区 (□)；c. 肾间组织 (★)；d. 内皮细胞 (→)和黑色素巨噬细胞中心 (▲)

Figure 2. HE Staining observation of head kidney of P. vachelli

a. Capsule (→), blood sinus (▲); b. lymphoid zone (○) and granulocytic zone (□); c. Anterior inter-renal tissue (★); d. Endothelial cells (→) and melanin macrophage centers (▲)

下载: 全尺寸图片幻灯片

图 3 瓦氏黄颡鱼正常头肾组织切片改良James染色观察

a．网状纤维覆盖于整个头肾 (→)；b. 分布于头肾被膜的网状纤维 (→)；c. 小静脉血管内侧及血管周围的网状纤维 (→)；d. 大动脉血管周围的胶原纤维 (→)；e. 黑色素巨噬细胞外层的网状纤维 (→)；f. 肾间组织内的网状纤维 (→)

Figure 3. Modified James staining observation of normal head kidney tissue sections of P. vachelli

a. Reticular fibers covered the entire head kidney (→); b. Reticular fibers (→) distributed in the cephalic renal capsule; c. Reticular fibers (→) inside and around venules; d. Collagen fibers around the aorta (→); e. Reticular fibers (→) on the outer layer of melanin macrophages; f. Reticular fibers (→) in anterior renal interstitials

下载: 全尺寸图片幻灯片

图 4 注射台盼蓝不同时间点瓦氏黄颡鱼体颜色变化

a. 正常体色，未注射台盼蓝；b. 注射第24小时；c. 注射第48小时；d. 注射第72小时

Figure 4. Changes of fish color at different time of injection trypan blue in P. vachelli

a. Normal body color without trypan blue injection; b. 24 h after injection; c. 48 h after injection; d. 72 h after injection

下载: 全尺寸图片幻灯片

图 5 瓦氏黄颡鱼头肾中台盼蓝的分布 (未染色)

a. 第0小时未注射台盼蓝，正常头肾；b. 第4小时台盼蓝开始出现在一些细胞里面，呈点状或小聚集团状 (→)；c. 第8小时台盼蓝开始增多(→)；d. 第24小时台盼蓝数量增多较明显，可明显看见小聚集团面积增大(→)；e. 第48小时台盼蓝增加特别明显，可看见大面积台盼蓝聚集团(→)；f. 第72小时台盼蓝数量继续增多(→)

Figure 5. Distribution of trypan blue in head kidney of P. vachelli (Unstained)

a. Trypan blue was not injected at the 0^th hour, normal head kidney; b. Trypan blue began to appear in some cells at the 4^th hour, showing punctate or small aggregates (→); c. Trypan blue began to increase at the 8^th hour (→); d. The number of Trypan blue increased more obviously at the 24^th hour, and the area of small aggregates could be clearly seen (→); e. The increase of Trypan blue at the 48^th hour was particularly obvious, and large area of Trypan blue aggregates could be seen (→); f. The number of trypan blue continued to increase at the 72^nd hour (→).

下载: 全尺寸图片幻灯片

图 6 瓦氏黄颡鱼头肾中第0至第72小时台盼蓝含量变化趋势

*. 0.01<P<0.05; **. P<0.01

Figure 6. Variation trend of trypan blue content in head kidney of P. vachelli from 0^th to 72^nd hour

下载: 全尺寸图片幻灯片

图 7 瓦氏黄颡鱼台盼蓝的分布部位

a. 第48小时头肾，HE染色，示台盼蓝(→)；b. a中同一区域头肾未染色，示台盼蓝(→)；c. 第72 小时头肾，HE染色，示台盼蓝(→)；d. c中同一区域头肾未染色，示台盼蓝(→)；e. 网状内皮细胞(→)；f. 血窦中游离的巨噬细胞(→)

Figure 7. Distribution of trypan blue in P. vachelli

a. 48^th hour head kidney, HE staining, showing trypan blue (→); b. The same area of a, unstained, showing trypan blue (→); c. 72^nd hour head kidney, HE staining, showing trypan blue (→); d. The same area of c, unstained, showing trypan blue (→); e. Reticular endothelial cells (→); f. Free macrophages in blood sinus (→)

下载: 全尺寸图片幻灯片

参考文献(33)

[1]	PRESS C M, EVENSEN Ø. The morphology of the immune system in teleost fishes[J]. Fish Shellfish Immunol, 1999, 9(4): 309-318. doi: 10.1006/fsim.1998.0181
[2]	BOEHM T, SWANN J B. Origin and evolution of adaptive immunity[J]. Annu Rev Anim Biosci, 2014, 2(1): 259-283. doi: 10.1146/annurev-animal-022513-114201
[3]	GEVEN E J, KLAREN P H. The teleost head kidney: integrating thyroid and immune signalling[J]. Dev Comp Immunol, 2017, 66: 73-83. doi: 10.1016/j.dci.2016.06.025
[4]	黄峰, 严安生. 黄颡鱼的含肉率及鱼肉营养评价[J]. 淡水渔业, 1999, 29(10): 3-6.
[5]	农业农村部渔业渔政管理局, 全国水产技术推广总站, 中国水产学会. 2019中国渔业统计年鉴[M]. 北京: 中国农业出版社, 2019: 24-25.
[6]	王德田, 董建强. 实用现代病理学技术[M]. 北京: 中国协和医科大学出版社, 2012: 86-97.
[7]	林鸿翔, 朱建善. 改良 James 染色法的应用及效果观察[J]. 上海交通大学学报 (医学版), 2009, 29(3): 359-360.
[8]	SINGH R, HUSSAIN M A, KUMAR J, et al. Chronic fluoride exposure exacerbates headkidney pathology and causes immune commotion in Clarias gariepinus[J]. Aquat Toxicol, 2017, 192: 30-39. doi: 10.1016/j.aquatox.2017.09.006
[9]	FISHELSON L. Cytomorphological alterations of the thymus, spleen, head-kidney, and liver in cardinal fish (Apogonidae, Teleostei) as bioindicators of stress[J]. J Morphol, 2006, 267(1): 57-69. doi: 10.1002/jmor.10385
[10]	何洁. 无乳链球菌对罗非鱼头肾损伤的病理学研究 [D]. 成都: 四川农业大学, 2018: 21-30.
[11]	BRUNO D W, NOGUERA P A, POPPE T T. A colour atlas of salmonid diseases [M]. New York: Springer Science & Business Media, 2013: 5-26.
[12]	刘云. 牙鲆免疫系统形态结构、发育和感染病理反应的研究[D]. 青岛: 中国海洋大学, 2003: 20-26.
[13]	罗俊标, 谢木娇, 区又君, 等. 四指马鲅头肾和脾脏组织学研究[J]. 生物学杂志, 2016, 33(4): 43-47. doi: 10.3969/j.issn.2095-1736.2016.04.043
[14]	李长玲, 曹伏君, 刘楚吾, 等. 花尾胡椒鲷脾脏和头肾显微结构的观察[J]. 海洋通报, 2002, 21(2): 30-35. doi: 10.3969/j.issn.1001-6392.2002.02.005
[15]	马燕梅, 林树根, 王全溪, 等. 花鲈头肾的显微结构和超显微结构[J]. 福建农林大学学报 (自然科学版), 2008, 37(2): 190-193.
[16]	区又君, 刘江华, 李加儿, 等. 驼背鲈头肾和脾脏的形态组织学观察[J]. 南方农业学报, 2015, 46(11): 2034-2039. doi: 10.3969/j:issn.2095-1191.2015.11.2034
[17]	胡玲玲, 李加儿, 区又君, 等. 条石鲷头肾和脾脏的显微结构观察[J]. 南方水产, 2010, 6(3): 41-45.
[18]	雷雪彬, 常藕琴, 石存斌, 等. 草鱼头肾发生组织学与免疫组织化学观察[J]. 水产学报, 2013, 37(6): 840-850.
[19]	王鸿鹄, 李长玲, 曹伏君, 等. 红笛鲷头肾和脾脏显微结构的观察[J]. 台湾海峽, 2006, 25(3): 360-367.
[20]	田敬云, 谢海侠, 姚卫建, 等. 鳜鱼头肾的组织发生及成鱼头肾B淋巴细胞的分布[J]. 动物学报, 2005, 51(3): 440-446. doi: 10.3969/j.issn.1674-5507.2005.03.009
[21]	岳兴建, 张耀光, 敖磊, 等. 南方鲇头肾的组织学和超微结构[J]. 动物学研究, 2004, 25(4): 327-333. doi: 10.3321/j.issn:0254-5853.2004.04.010
[22]	曹守花, 李加儿, 区又君, 等. 鲻脾脏和头肾的形态学与组织学研究[J]. 海洋渔业, 2013, 35(3): 249-256. doi: 10.3969/j.issn.1004-2490.2013.03.001
[23]	龚宁, 聂其灼. 鲤鱼头肾显微结构研究[J]. 山地农业生物学报, 2001, 20(1): 20-24. doi: 10.3969/j.issn.1008-0457.2001.01.004
[24]	祁保霞. 黄颡鱼脾脏和头肾的显微结构观察[J]. 內蒙古民族大学学报 (自然科学版), 2010, 25(3): 304-306.
[25]	钟明超, 罗泉笙. 中国鲿科鲶类肾的外形观察[J]. 西南师范大学学报 (自然科学版), 1991(2): 71-77.
[26]	MACHADO M R F, SOUZA H D O, SOUZA V L D, et al. Morphological and anatomical characterization of the digestive tract of Centropomus parallelus and Centropomus undecimalis[J]. Acta Scientiarum Biol Sci, 2013, 35(4): 467-474.
[27]	胡宗福, 尹殿明, 温海深, 等. 条斑星鲽肾脏的组织学观察[J]. 内蒙古民族大学学报 (自然科学版), 2011(1): 39-42.
[28]	KLEI T R, MEINDERTS S M, van den BERG T K, et al. From the cradle to the grave: the role of macrophages in erythropoiesis and erythrophagocytosis[J]. Front Immunol, 2017, 8: 73. doi: 10.3389/fimmu.2017.00073
[29]	SAYED A H, YOUNES H A. Melanomacrophage centers in Clarias gariepinus as an immunological biomarker for toxicity of silver nanoparticles[J]. J Microsc Ultrastruct, 2017, 5(2): 97-104. doi: 10.1016/j.jmau.2016.07.003
[30]	WOLKE R, GEORGE C, BLAZER V. Pigmented macrophage accumulations (MMC; PMB): possible monitors of fish health[J]. Parasitol Pathol Mar Organ World Ocean, 1985, 25: 93-97.
[31]	KUMAR R, JOY K P, SINGH S M. Morpho-histology of head kidney of female catfish Heteropneustes fossilis: seasonal variations in melano-macrophage centers, melanin contents and effects of lipopolysaccharide and dexamethasone on melanins[J]. Fish Physiol Biochem, 2016, 42(5): 1287-1306. doi: 10.1007/s10695-016-0218-2
[32]	HEINING P, HOFFMANN R W. Light-and electron microscopical studies on the prolonged toxicity of trichloroethylene on rainbow trout (Oncorhynchus mykiss)[J]. Exp Toxicol Pathol, 1993, 45(2/3): 167-176.
[33]	贺扬. 感染无乳链球菌尼罗罗非鱼脾脏的病理学研究 [D]. 成都: 四川农业大学, 2017: 19-38.