温度与脂多糖对栉孔扇贝血细胞吞噬活力的影响

Effect of Water Temperature on Phagocytic Activitiy of the Haemocytes of Scallop(Chlamys farreri)After Stimulated by Lipopolysaccharide

贝类无特异性免疫系统,细胞吞噬是其免疫防御的重要方式之一.在5、10、15、20和25℃养殖条件下,给栉孔扇贝（Chlamysfarreri）分别注射荧光微球、脂多糖和荧光微球混合液.在注射后的0,6,12,18,24,30,36,42和48 h抽取血细胞,用荧光标记血细胞以观察其对微球的吞噬状态,用流式细胞术测定血细胞对微球的吞噬率.结果表明,在5个养殖温度下,2个处理组均自6h后扇贝血细胞内即出现吞噬的微球,之后吞噬率逐渐上升,直到18～24 h达到最高峰,随后逐渐下降.5、10、15、20和25℃条件下,注射荧光微球组血细胞最大吞噬率分别为8.66％±0.26％、14.00％±0.22％、16.97％±0.55％、25.62％±0.88％和8.97％±0.25％,注射脂多糖和荧光微球混合液组分别血细胞最大吞噬率为9.75％±0.48％、17.49％±0.43％、23.13％±0.57％、34.16％±0.97％和10.42％±0.45％.20℃组的两处理组血细胞吞噬率显著高于其他温度组,其次是15和10℃组,最低是5和25℃组,并且20、15℃组的最大吞噬率出现在18h,其他组均在24h.各养殖温度组中,脂多糖和荧光微球混合液组血细胞吞噬率均高于荧光微球组.研究表明：栉孔扇贝血细胞的吞噬活力受水温影响显著,脂多糖能够显著增强扇贝血细胞的吞噬活力.

Phagocytosis of haemocytes is the primary innate immune defense of scallop （Chlamys farreri）.In this study,the scallop was accustomed at 5,10,15,20 and 25 ℃,respectively,then injected with either phycoerythrin labeled fluorescent microparticles （FM）,lipopolysaccharide （LPS） plus FM or PBS.After that,haemocytes were collected at 0,6,12,18,24,30,36,42 and 48 hours post injection,respectively,and their phagocytic activity was measured with fluorescence microscope and flow cytometry （FCM）.The results showed that haemocytes engulfed the FM at 6h,and the engulfing rate rose in the first 18-24 hours,reaching the maximum and then gradually declined.At 5 ℃,the maximum of haemocyte engulfing rate in the two treatment were 8.66％ ± 0.26％ and 9.75％ ± 0.48％,respective;at 10 ℃ they were 14.00％ ± 0.22％ and 17.49％ ± 0.43％;at 15 ℃,they were 16.97％ ± 0.55％ and 23.13％± 0.57％;at 20 ℃ were 25.62％ ± 0.88％ and 34.16％ ± 0.97％;at 25 ℃,they were 8.97 ％ ± 0.25％ and 10.42％ ± 0.45％.Engulfing rate in LPS-treated group was significantly higher than that in FM group.Moreover,at 20 ℃,engulfing rate was significantly higher than that of other groups,followed by 15 and 10 ℃ groups,and finally 5 and 25 ℃ groups,No significant difference between groups at 15 and 20 ℃.The results suggested that water temperature affected the engulfing rate significantly and LPS significantly enhanced the phagocytic activity of scallop haemocytes.