N乙酰D氨基葡糖体外对小鼠淋巴细胞增殖和细胞内钙调蛋白和钙调神经磷酸酶表达影响.docVIP

N乙酰D氨基葡糖体外对小鼠淋巴细胞增殖和细胞内钙调蛋白和钙调神经磷酸酶表达影响 　　摘要：目的研究N-乙酰-D-氨基葡糖（N-AcGA）对小鼠淋巴细胞的增殖作用及对细胞内钙调蛋白（CaM）和钙调神经磷酸酶（CaN）表达的影响。方法以小鼠淋巴细胞为实验材料，通过MTT法及流式细胞仪研究N-AcGA对小鼠淋巴细胞增殖和对淋巴细胞内CaM与CaN表达的影响。结果N-AcGA在200～50 mg/L浓度范围内对小鼠淋巴细胞增殖有显著的促进作用，浓度为200 mg/L时作用极为显著。N-AcGA可增加小鼠淋巴细胞内CaM和CaN的表达。加入N-AcGA 72 h后CaM的平均阳性细胞百分率由61.6%上升到72.6%，CaN的平均阳性细胞百分率由75.6%上升到86.0%。结论N-AcGA能够诱导小鼠淋巴细胞增殖，并促进细胞内CaM与CaN的表达。 　　关键词：N-乙酰-D-氨基葡糖；淋巴细胞增殖；钙调蛋白；钙调神经磷酸酶；小鼠 　　中图分类号：R977.6文献标识码：A文章编号：1672-979X(2007)07-0013-03 　　Effects of N-Acetyl-D-Glucosamine on Lymphocyte Proliferation of Mice in Vitro and Expression of Intracellular CaM and CaN 　　LI Hui, CAO Xiu-ming, JI Yu-bin, ZHANG Zhen-zhu 　　(Center of Research on Life Sciences and Environmental Sciences, Harbin University of Commerce, Harbin 150076, China) 　　Abstract：Objective To investigate the effects of N-acetyl-D-glucosamine（N-AcGA）on the lymphocyte proliferation of mice and the expression of intracellular CaM and CaN. Methods The effect of N-AcGA on lymphocyte proliferation was observed by MTT. Flow cytometry was used to detect the expression of intracellular CaM and CaN. Results N-AcGA promoted lymphocyte proliferation of mice when the concentration was 200~50 mg/L in substrate and its effect was especially marked when the concentration was 200 mg/L. N-AcGA could increase the expression of intracellular CaM and CaN. The average positive cells percentage of CaM raised from 61.6% to 72.6% ｗhen adding N-AcGA after 72 h, while that of CaN raisedfrom 75.6% to 86.0%. Conclusion N-AcGA can stimulate the lymphocyte proliferation and the expression of intracellular CaM and CaN. 　　Key words：N-acetyl-D-glucosamine; lymphocyte proliferation; calmodulin; calcineurin; mice 　　N-乙酰D-氨基葡萄糖(N-Acetyl-D-Glucosamine，N-AcGA)是甲壳素的基本组成单位，具有消炎、抗肿瘤及抗氧化等作用，是治疗骨关节炎及风湿性关节炎的有效药物[1]。钙调蛋白（calmodulin, CaM）是Ca2+信号的主要调节蛋白或受体蛋白。而钙调神经磷酸酶（calcineurin, CaN）是在细胞信号传递中直接受Ca2+调节的关键酶，具有激活体内T淋巴细胞的作用。本研究采用MTT（溴化四唑蓝）法及流式细胞仪观察了N-AcGA对小鼠淋巴细胞的增殖和对淋巴细胞内CaM和CaN表达的影响。 　　 　　1材料 　　 　　1.1药品与试剂 　　N-AcGA（纯