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CLC number: Q25

On-line Access: 2015-12-04

Received: 2015-04-25

Revision Accepted: 2015-09-14

Crosschecked: 2015-11-24

Cited: 4

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Citations:  Bibtex RefMan EndNote GB/T7714


Hai-sheng Xu


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Journal of Zhejiang University SCIENCE B 2015 Vol.16 No.12 P.971-979


Effect of lipopolysaccharide on the hemocyte apoptosis of Eriocheir sinensis

Author(s):  Hai-sheng Xu, Sun-jian Lyu, Jie-hao Xu, Bin-jie Lu, Jing Zhao, Song Li, Yi-qun Li, Yu-yin Chen

Affiliation(s):  College of Animal Sciences, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   hsxu@zju.edu.cn, chenyy@zju.edu.cn

Key Words:  Hemocyte, Eriocheir sinensis, Lipopolysaccharide, Apoptosis, Reactive oxygen species

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Hai-sheng Xu, Sun-jian Lyu, Jie-hao Xu, Bin-jie Lu, Jing Zhao, Song Li, Yi-qun Li, Yu-yin Chen. Effect of lipopolysaccharide on the hemocyte apoptosis of Eriocheir sinensis[J]. Journal of Zhejiang University Science B, 2015, 16(12): 971-979.

@article{title="Effect of lipopolysaccharide on the hemocyte apoptosis of Eriocheir sinensis",
author="Hai-sheng Xu, Sun-jian Lyu, Jie-hao Xu, Bin-jie Lu, Jing Zhao, Song Li, Yi-qun Li, Yu-yin Chen",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Effect of lipopolysaccharide on the hemocyte apoptosis of Eriocheir sinensis
%A Hai-sheng Xu
%A Sun-jian Lyu
%A Jie-hao Xu
%A Bin-jie Lu
%A Jing Zhao
%A Song Li
%A Yi-qun Li
%A Yu-yin Chen
%J Journal of Zhejiang University SCIENCE B
%V 16
%N 12
%P 971-979
%@ 1673-1581
%D 2015
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1500098

T1 - Effect of lipopolysaccharide on the hemocyte apoptosis of Eriocheir sinensis
A1 - Hai-sheng Xu
A1 - Sun-jian Lyu
A1 - Jie-hao Xu
A1 - Bin-jie Lu
A1 - Jing Zhao
A1 - Song Li
A1 - Yi-qun Li
A1 - Yu-yin Chen
J0 - Journal of Zhejiang University Science B
VL - 16
IS - 12
SP - 971
EP - 979
%@ 1673-1581
Y1 - 2015
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1500098

In the present study, we investigated the possible toxicity mechanism of lipopolysaccharide (LPS) extracted from Gram-negative bacteria in Eriocheir sinensis hemocytes. Apoptotic hemocytes and reactive oxygen species (ROS) production induced by the LPS were monitored by the combination of flow cytometry and microscope observation. It was shown that LPS induced serious damage on the DNA and morphological changes in hemocytes, including cell shrinkage, fracture of nucleus membrane, margination, condensation and fragmentation of chromatin, and formation of apoptotic bodies indicating obvious hemocyte apoptosis. As compared with the control group, the apoptotic cell ratio increased to 30.61% and 39.01% after 1-h exposure and 57.72% and 75.01% after 2-h exposure to 1 and 10 μg/ml LPS, respectively (P<0.05). Significant outburst of ROS production was observed in LPS-treated hemocytes with approximately 176.6% of relative dichlorofluorescein mean fluorescence at 1-h exposure, followed by a drastic decline (P<0.05). These results indicated that LPS would induce oxidative stress on hemocytes from E. sinensis and cause ROS burst, DNA damage, and subsequently apoptosis. The process of ROS-mediated apoptosis might be one of the potential toxicity mechanisms of LPS on crustacean hemocytes.


方法:在体外26 °C条件下,培养中华绒螯蟹血细胞。用浓度1 µg/ml 的LPS处理血细胞0、4、8和16 h后,利用4’,6-二脒基-2-苯基吲哚(DAPI)染色、荧光显微镜和透射电子显微镜观察血细胞的形态变化,用DNA Ladder试剂盒检测脱氧核糖核酸(DNA)的损伤程度。分别用浓度1和10 µg/ml的LPS处理血细胞0、1和2 h,经膜联蛋白 V-异硫氰酸荧光素(V-FITC)/碘化丙啶(PI)染色后,用流式细胞仪检测血细胞凋亡数量。收集血细胞制成细胞悬浮液,加入LPS至终浓度为10 µg/ml,同时加入2’,7’-二氯荧光黄双乙酸盐(DCFH-DA)探针至终浓度为1 µmol/L;对照组加入等量生理盐水和DCFH-DA探针。在26 °C孵育不同时间后,用流式细胞仪和激光共聚焦显微镜检测血细胞活性氧的产生情况。


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Brinkmann, V., Zychlinsky, A., 2012. Neutrophil extracellular traps: is immunity the second function of chromatin? J. Cell Biol., 198(5):773-783.

[2]Chang, C.C., Yeh, M.S., Lin, H.K., et al., 2008. The effect of Vibrio alginolyticus infection on caspase-3 expression and activity in white shrimp Litopenaeus vannamei. Fish Shellfish Immunol., 25(5):672-678.

[3]Chang, M., Wang, W.N., Wang, A.L., et al., 2009. Effects of cadmium on respiratory burst, intracellular Ca2+ and DNA damage in the white shrimp Litopenaeus vannamei. Comp. Biochem. Phys. C, 149(4):581-586.

[4]Cheng, W., Chen, J.C., 2001. Effects of intrinsic and extrinsic factors on the haemocyte profile of the prawn, Macrobrachium rosenbergii. Fish Shellfish Immunol., 11(1):53-63.

[5]Du, J., Zhu, H., Ren, Q., et al., 2012. Flow cytometry studies on the Macrobrachium rosenbergii hemocytes sub-populations and immune responses to novel pathogen spiroplasma MR-1008. Fish Shellfish Immunol., 33(4):795-800.

[6]Gai, Y., Zhao, J., Song, L., et al., 2008. A prophenoloxidase from the Chinese mitten crab Eriocheir sinensis: gene cloning, expression and activity analysis. Fish Shellfish Immunol., 24(2):156-167.

[7]Jiravanichpaisal, P., Lee, B.L., Söderhäll, K., 2006. Cell-mediated immunity in arthropods: hematopoiesis, coagulation, melanization and opsonization. Immunobiology, 211(4):213-236.

[8]Johansson, M.W., Söderhäll, K., 1985. Exocytosis of the prophenoloxidase activating system from crayfish haemocytes. J. Comp. Physiol. B, 156(2):175-181.

[9]Johansson, M., Soderhall, K., 1989. Cellular immunity in crustaceans and the proPO system. Parasitol. Today, 5(6):171-176.

[10]Johansson, M.W., Keyser, P., Sritunyalucksana, K., et al., 2000. Crustacean haemocytes and haematopoiesis. Aquaculture, 191(1):45-52.

[11]Khanobdee, K., Soowannayan, C., Flegel, T.W., et al., 2002. Evidence for apoptosis correlated with mortality in the giant black tiger shrimp Penaeus monodon infected with yellow head virus. Dis. Aquat. Org., 48(2):79-90.

[12]le Moullac, G., Haffner, P., 2000. Environmental factors affecting immune responses in Crustacea. Aquaculture, 191(1-3):121-131.

[13]Li, C.C., Chen, J.C., 2008. The immune response of white shrimp Litopenaeus vannamei and its susceptibility to Vibrio alginolyticus under low and high pH stress. Fish Shellfish Immunol., 25(6):701-709.

[14]Lorenzon, S., de Guarrini, S., Smith, V.J., et al., 1999. Effects of LPS injection on circulating haemocytes in crustaceans in vivo. Fish Shellfish Immunol., 9(1):31-50.

[15]Lv, S.J., Xu, J.H., Zhao, J., et al., 2014. Classification and phagocytosis of circulating haemocytes in Chinese mitten crab (Eriocheir sinensis) and the effect of extrinsic stimulation on circulating haemocytes in vivo. Fish Shellfish Immunol., 39(2):415-422.

[16]Lv, S.J., Lu, B.J., Xu, J.H., et al., 2015. Immune response of peroxinectin of Chinese mitten crab Eriocheir sinensis to exterior stimulation. Dev. Comp. Immunol., 51(1):56-64.

[17]Orrenius, S., 2007. Reactive oxygen species in mitochondria-mediated cell death. Drug Metab. Rev., 39(2-3):443-455.

[18]Persson, M., Cerenius, L., Söderhäll, K., 1987. The influence of haemocyte number on the resistance of the freshwater crayfish, Pacifastacus leniusculus Dana, to the parasitic fungus Aphanomyces astaci. J. Fish Dis., 10(6):471-477.

[19]Pijanowski, L., Golbach, L., Kolaczkowska, E., et al., 2013. Carp neutrophilic granulocytes form extracellular traps via ROS-dependent and independent pathways. Fish Shellfish Immunol., 34(5):1244-1252.

[20]Portt, L., Norman, G., Clapp, C., et al., 2011. Anti-apoptosis and cell survival: a review. Biochim. Biophys. Acta, 1813(1):238-259.

[21]Raman, T., Arumugam, M., Mullainadhan, P., 2008. Agglutinin-mediated phagocytosis-associated generation of superoxide anion and nitric oxide by the hemocytes of the giant freshwater prawn Macrobrachium rosenbergii. Fish Shellfish Immunol., 24(3):337-345.

[22]Sahtout, A.H., Hassan, M., Shariff, M., 2001. DNA fragmentation, an indicator of apoptosis, in cultured black tiger shrimp Penaeus monodon infected with white spot syndrome virus (WSSV). Dis. Aquat. Org., 44(2):155-159.

[23]van de Braak, C.B.T., Botterblom, M.H.A., Liu, W., et al., 2002. The role of the haematopoietic tissue in haemocyte production and maturation in the black tiger shrimp (Penaeus monodon). Fish Shellfish Immunol., 12(3):253-272.

[24]Xian, J.A., Wang, A.L., Ye, C.X., et al., 2010. Phagocytic activity, respiratory burst, cytoplasmic free-Ca2+ concentration and apoptotic cell ratio of haemocytes from the black tiger shrimp, Penaeus monodon under acute copper stress. Comp. Biochem. Phys. C, 152(2):182-188.

[25]Xian, J.A., Wang, A.L., Chen, X.D., et al., 2011. Cytotoxicity of nitrite on haemocytes of the tiger shrimp, Penaeus monodon, using flow cytometric analysis. Aquaculture, 317(1):240-244.

[26]Xian, J.A., Miao, Y.T., Li, B., et al., 2013. Apoptosis of tiger shrimp (Penaeus monodon) haemocytes induced by Escherichia coli lipopolysaccharide. Comp. Biochem. Phys. A, 164(2):301-306.

[27]Zhang, Y., Zhao, J.M., Zhang, H., et al., 2010. The involvement of suppressors of cytokine signaling 2 (SOCS2) in immune defense responses of Chinese mitten crab Eriocheir sinensis. Dev. Comp. Immunol., 34(1):42-48.

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