大豆凝集素对鲤鱼肠道上皮细胞结构和功能的影响

摘要

This experiment was conducted to study the influence of soybean agglutinin (SB A) on the structure and function of carp intestinal epithelial cells in primary culture. The carp intestinal epithelial cells in primary culture were cultured in 24-well plate after 72 h, then changed into the DMEM culture medium which contained 0, 50, 75, 100, 150 and 200 μg/mL purified SBA, respectively. The samples were collected to measure the indices after 72 h SBA culture. The results showed that the activities of lactate dehydrogenase (LDH) and glutamic-pyruvic transaminase (GPT) in culture medium of carp intestinal epithelial cells were significantly enhanced by SBA (P <0. 01) , but the influence on glutamic-oxalacetic transaminase (GOT) activity in the culture medium didn't reach significant level (P > 0. 05). The results of cell morphological observation showed that the cell shape was obviously changed by SBA, and it was changed from ellipse to long strip or irregular shape and the number of dead cells in culture medium increased. Meanwhile, the SBA could significantly increase the methyl thiazolyl tetrazolium optical density (MTT OD) value and protein content (P <0. 05 or P < 0. 01) in carp intestinal epithelial cells, and significantly enhance the activities of GOP and GPT in cells and ammonia concentration in culture medium (P <0.05 or P<0.01), but significantly decrease the activities of alkaline phosphatase (AKP) and Na+ ,K + -ATPase in cells (P <0. 01). These results indicate that SBA can damage the normal structure of cells, promote the proliferation and apoptosis of cells, restrain cell differentiation and accelerate the protein metabolism for carp intestinal epithelial cells.%本研究旨在考察大豆凝集素(SBA)对原代培养鲤鱼肠道上皮细胞结构和功能的影响,初步探讨大豆凝集素在鱼类中抗营养的可能作用方式.在24孔培养板中原代培养鲤鱼肠道上皮细胞72 h后,分别更换为含纯化sBA 0、50、75、100、150、200μg/mL的DMEM培养液,继续培养72 h后采样测定指标.结果表明,在SBA的作用下,鲤鱼肠道上皮细胞培养液中乳酸脱氢酶(LDH)、谷丙转氨酶(GPT)活力极显著增加(P＜0.01),但培养液中谷草转氨酶(GOT)活力变化不显著(P＞0.05).细胞形态学观察表明,sBA使细胞形态发生了明显变化,细胞由原有的椭圆形变成长条形或不规则形,培养液中漂浮的死细胞数量增多.同时,SBA显著或极显著增加鲤鱼肠道上皮细胞中四甲基偶氮唑盐(MTT)OD值、蛋白质含量(P＜0.05或P＜0.01),显著或极显著增加细胞中GPT和GOT活力及培养液中氨浓度(P＜0.05或P＜0.01);极显著降低细胞中碱性磷酸酶(AKP)和Na+,K+-ATP酶活力(P＜0.01).由此得出,SBA破坏了鲤鱼肠道上皮细胞结构的完整性,促进了细胞的增殖和凋亡,抑制了细胞的成熟分化,加速了细胞的蛋白质代谢.