中华绒螯蟹(Eriocheir sinensis)血细胞生成的初步研究

A preliminary study on hemopoiesis of Eriocheir sinensis

为了揭示失血和LPS刺激后中华绒螯蟹血细胞的生成及成熟过程,利用流式细胞技术将中华绒螯蟹造血组织细胞和血细胞按大小进行分类,利用EdU标记检测失血刺激后中华绒螯蟹造血组织细胞的增殖情况,分析失血和LPS刺激后中华绒螯蟹不同类型血细胞比例的变化.结果显示:中华绒螯蟹造血组织细胞分布于106~384μm^(2)之间,而血细胞分布在119~434μm^(2)之间,其中,透明细胞和小颗粒细胞主要分布在200~300μm^(2)之间,中颗粒和大颗粒细胞主要分布在300~400μm^(2)之间,随着血细胞颗粒性的增加,细胞逐渐增大.在失血刺激6、12和24 h时,中华绒螯蟹造血组织细胞增殖加速,说明失血刺激可以促进造血组织细胞增殖以加速新血细胞的生成.失血刺激2 h后透明细胞比例显著降低,至12 h时,各类细胞基本恢复至正常水平.LPS刺激1、3、6和9 h后,透明细胞比例呈现先降低后升高再降低的变化,而小、中和大颗粒细胞比例变化与透明细胞比例变化趋势相反,LPS刺激24 h和48 h后,透明细胞比例显著升高,大颗粒细胞比例显著降低.由此可知,失血和LPS刺激引起了中华绒螯蟹机体不同细胞的免疫反应,可能通过不同方式影响血细胞的生成、分化和成熟.

In order to reveal the process of formation and maturation of hemocytes in Eriocheir sinensis after haemolymph loss and LPS stimulation,the hemopoietic tissue cells and hemocytes of E.sinensis were classified and analyzed according to their size by flow cytometry.EdU labeling was used to detect the proliferation of hemopoietic tissue cells in E.sinensis after haemo-lymph loss,and the proportion of different types of hemocyte of E.sinensis was analyzed after haemolymph loss and LPS stimulation.The results showed that the hemopoietic tissue cells of E.sinensis were distributed in the range of 106-384μm^(2),while the hemocytes were distributed in the range of 119-434μm^(2),among which the hyaline cells and small granular cells were mainly distributed in the range of 200-300μm^(2),the medium granular and large granular cells were mainly distributed in the range of 300-400μm^(2),and the cells gradually increased with the increase of hemocyte granularity.The proliferation of hemopoietic tissue cells was found to be increasing at 6,12 and 24 h after haemolymph loss stimulation,indicating that haemolymph loss could promote the proliferation of hemopoietic tissue cells to accelerate the generation of new hemocytes.The proportion of hyaline cells decreased significantly after 2 h of haemolymph loss stimulation,and then returned to the nor-mal level at 12 h.The proportion of hyaline cells significantly decreased at first,then increased,and then decreased after 1,3,6 and 9 h of LPS stimulation,while the proportion of small,medium and large granular cells changed in the opposite way to the proportion of hyaline cells.After 24 h and 48 h of LPS stimulation,the proportion of hyaline cells was significantly higher and the proportion of large granular cells was significantly lower.It is concluded that haemolymph loss and LPS stimulation elicited different cellular immune responses in the body of E.sinensis,which may affect the formation,differentiation and maturation of hemocytes in different ways.