高糖环境下美罗华对B淋巴细胞株Ramos增殖的抑制作用  

High glucose levels enhanced the inhibitory effects of rituximab on the proliferation of Ramos B lymphocytes

作　　者：陈樱[1] 孟东[1] 金秋颖[2] 张捷[1] 邓为民[2] 李海东[2] 

机构地区：[1]天津医科大学代谢病医院,300070 [2]天津医科大学基础医学院

出　　处：《中华糖尿病杂志》2012年第7期430-433,共4页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家自然科学基金资助项目（81070271）

摘　　要：目的探讨在高糖状态下美罗华对B淋巴细胞周期、增殖和凋亡的影响。方法体外培养人B淋巴细胞株Ramos，将细胞分别以单纯5．5、10．0、15．0、25．0mmol/L葡萄糖培养或另外分别加入10mg／L美罗华培养，即分为葡萄糖组和美罗华组。以上两组培养3—7d，检测细胞增殖、凋亡及其周期情况。细胞增殖和活性以台盼蓝染色法测定。细胞凋亡和周期采用流式细胞技术检测。两组间数据比较采用配对t检验。结果5．5、10．0、15．0、25．0mmol/L葡萄糖组B淋巴细胞数量分别为（1．96±0．41）×10^6、（3．49±0．51）×10^6、（3．44±0．67）×10^6、（3．04±0．52）×10^6，凋亡率分别为5．0％±0．9％、4．0％±0．8％、6．2％±1．8％、7．6％±1．3％，S期比例分别为32％±6％、41％±8％、40％±7％、36％±6％；美罗华组细胞数量分别为（1．23±0．23）×10^6、（1．52±0．29）×10^6、（1．38±0．23）××10^6、（1．06±0．23）×10^6，细胞凋亡率则分别23．1％±3．2％、21．2％±4．2％、24．4％±4．8％、26．9％±6．0％，S期细胞比例则分别22％±4％、28％±5％、26％±4％、20％±5％。与葡萄糖组比较，美罗华组B细胞增殖在不同糖浓度亚组均显著减少（t=9．19、5．52、9．75、14．22，均P〈0．01），细胞凋亡率显著增加（t：16．62、12．51、11．53、9．12，均P〈0．01），S期细胞比率显著减少（t=5．87、5．19、9．91、7．73，均P〈0．01）。两组细胞增殖数量和S期比率均在10．0mmol/L葡萄糖时最高；两组细胞凋亡率均在25．0mmol/L葡萄糖时最多。结论美罗华通过抑制细胞周期以及促进细胞凋亡实现对B淋巴细胞增殖的抑制作用，其抑制作用在高糖状态下更为明显。Objective To investigate the effect of rituximab on cell cycle, proliferation and apoptosis of Ramos B lymphocytes in the presence of high levels of glucose. Methods Ramos B lymphocytes were treated with or without rituximab in the presence of different concentrations of glucose （5.5, 10. 0, 15.0 and 25.0 mmol/L）（ Glucose group and Rituximab group）. The cells were cultured for 3 -7 days. Cell cycle and apoptosis were studied by flow cytometry. Cell proliferation was measured by counting the cells using a hemoeytometer after trypan blue staining. Paired t test was applied when data were compared between the two groups. Results The numbers of B cells in 5.5, 10. 0, 15.0 and 25.0 mmoL/L glucose groups were （1.96 ±0.41） ×10^6, （3.49±0.51） ×10^6, （3.44 ±0.67）×10^6 and （3.04 ± 0. 52） ×10^6, respectively; the percentages of apoptotic cells were 5.0% ± 0. 9%, 4. 0% ± 0. 8%, 6. 2% ± 1.8% and 7.6% ± 1.3% ,respectively; the percentages of cells in S phase were 32% ±6%, 41% ±8%, 40% ± 7% and 36% ± 6%, respectively. The numbers of B cells in rituximab groups with increasing concentrations of glucose were （ 1.23 ± 0. 23 ）×10^6, （ 1.52 ± 0. 29 ） ×10^6, （ 1.38 ± 0. 23 ）×10^6 and （ 1.06 ± 0. 23 ） ×10^6 , respectively; the percentages of apoptotic cells were 23.1% ± 3.2% , 21.2% ±4. 2% , 24. 4% ± 4. 8% and 26. 9% ± 6. 0% , respectively; the percentages of cells in S phase were 22% ±4%, 28% ±5%, 26% ±4% and 20% ±5%, respectively. Compared with those in glucose group, the B cell proliferation was significantly inhibited （ t = 9. 19, 5.52, 9. 75, 14. 22, all P 〈 0. 01 ）, the apoptosis rates were significantly raised （ t = 16. 62, 12. 51, 11.53, 9. 12, all P 〈 0. 01 ） , and the percentage of cells in S phase increased significantly （ t = 5.87, 5. 19, 9.91, 7.73, all P 〈 0. 01 ） in Rituximab group. Highest cellular proliferation and percentage of cells in S phase were observed in the presence of 10. 0 mmol/L glucose. Highest rate of apoptosis oc

关 键 词：美罗华 高糖 凋亡 胰腺Β细胞 B淋巴细胞 

分 类 号：R965[医药卫生—药理学]