通过渗透微泵稳定抑制小鼠血浆CD26/DPPIV分子酶活性的研究

doi:10.3969/j.issn.1006-7795.2019.03.018

首都医科大学学报 ›› 2019, Vol. 40 ›› Issue (3): 424-428.doi: 10.3969/j.issn.1006-7795.2019.03.018

通过渗透微泵稳定抑制小鼠血浆CD26/DPPIV分子酶活性的研究

薛兴奎¹, 韩新华², 蔡思齐^1,3, 陈晓玲¹, Melody Y. Zeng¹

1. 深圳市龙华区人民医院中心实验室, 广东深圳 518109;
2. 深圳市南山区蛇口人民医院内分泌科, 广东深圳 518109;
3. 南方医科大学研究生院检验技术系, 广州 501505

收稿日期:2019-02-15 出版日期:2019-05-21 发布日期:2019-06-13
通讯作者: 薛兴奎 E-mail:xuexk@outlook.com
基金资助:
国家自然科学基金（31650110474），深圳市科学技术创新委员会（JCYJ20160426094752965），深圳市卫生系统科研项目（201601059），深圳市龙华区科技创新项目（20160831A1030180，20160831A0410020）。

Investigation on the inhibition of Sitagliptin by osmotic pump on CD26/DPPIV activity

Xue Xingkui¹, Han Xinhua², Cai Siqi^1,3, Chen Xiaoling¹, Melody Y. Zeng¹

1. Central Lab, The People's Hospital of Longhua Shenzhen, Shenzhen 518109, Guangdong Province, China;
2. Department of Endocrinology, Shenzhen Shekou People's Hospital, Shenzhen 518109, Guangdong Province, China;
3. Department of Laboratory Medicine and Biotechnology, Graduate School, Southern Medical University, Guangzhou 501505, China

Received:2019-02-15 Online:2019-05-21 Published:2019-06-13
Supported by:
This study was supported by National Natural Sciences Foundation of China (31650110474),Shenzhen Science,Technology and Innovation Commission (JCYJ20160426094752965),Shenzhen Health Research Project (201601059),Funds for Innovation of Science and Technology in Longhua District,Shenzhen (20160831A1030180,20160831A0410020).

摘要/Abstract

摘要： 目的研究采用皮下植入渗透微泵给予西他列汀对小鼠体内CD26/DDPIV蛋白酶活性的作用效果与持续时间。方法将灌注西他列汀药物的渗透泵经手术植入BL6/C57小鼠皮下，通过高效液相色谱-质谱监测小鼠体内血液药物浓度，酶标法检测小鼠血浆DPPIV蛋白酶活性，流式细胞术观察小鼠脾T细胞CD26分子的表达，利用Transwell小室检测小鼠T细胞迁移能力。取渗透泵周围皮肤，病理切片检查小鼠皮下病理损害或炎性反应。结果 B6/C57小鼠在植入渗透泵后，体内药物浓度比采用直接皮下注射法稳定，持续时间久，血浆DPPIV蛋白酶活性抑制程度与血药浓度明显相关。给药24 h后，小鼠脾T细胞CD26分子表达无明显变化，渗透泵组迁移能力较对照组明显降低。皮下植入渗透泵小鼠组饮食活动度无明显改变，皮肤病理检查未见炎性反应与病理损害。结论通过渗透泵给予小鼠药物能够保持西他列汀血药浓度稳定，持续抑制体内DPPIV活性，降低T细胞迁移能力，为CD26/DPPIV分子功能与免疫体内研究提供更方便有效的途径。

关键词: CD26/DPPIV分子, 西他列汀, 渗透泵, T细胞

Abstract: Objective To investigate the effect of Sitagliptin on mice with subcutaneous osmotic pump. Methods Osmotic pump carrying with Sitaglipin was implanted into mice subcutaneously, and the plasma concentration of sitagliptin was measured with HPLC-MS. The skin inflammation reaction was evaluated with pathological slides. DPPIV enzyme activity was measured with ELISA. Furthermore, the CD26 expression of spleen T cell was detected with flow cytometry and T cell migration was determined with transwell system.Results The duration of plasm Sitagliptin in mice subject to instant shot was shorter while those delivered by osmotic pump was observed at 24h post implantation of osmotic pump. Inhibition of DPPIV activity, as measured by ELISA, was confirmed at 24 hours after pump implantation, highlighting the utility of this method as a longer term model for DPPIV inhibition in mice. T cell migration was stably inhibited after application of Sitagliptin with osmotic pump. Conclusion In vivo Sitagliptin administration by an osmotic pump offers an effective approach for longer-term and stable DPPIV inhibition in mice and avoids the necessity for multiple dosing of the inhibitor, making study on function of CD26/DPPIV more effective.

Key words: CD26/DPPIV, Sitagliptin, osmotic pump, T cells

中图分类号:

R363

薛兴奎, 韩新华, 蔡思齐, 陈晓玲, Melody Y. Zeng. 通过渗透微泵稳定抑制小鼠血浆CD26/DPPIV分子酶活性的研究[J]. 首都医科大学学报, 2019, 40(3): 424-428.

Xue Xingkui, Han Xinhua, Cai Siqi, Chen Xiaoling, Melody Y. Zeng. Investigation on the inhibition of Sitagliptin by osmotic pump on CD26/DPPIV activity[J]. Journal of Capital Medical University, 2019, 40(3): 424-428.

参考文献

[1] Ohnuma K, Hatano R, Komiya E, et al. A novel role for CD26/dipeptidyl peptidase IV as a therapeutic target[J]. Front Biosci (Landmark Ed), 2018, 23:1754-1779.
[2] Farag S S, Srivastava S, Messina-Graham S, et al. In vivo DPP-4 inhibition to enhance engraftment of single-unit cord blood transplants in adults with hematological malignancies[J]. Stem Cells Dev, 2013, 22(7):1007-1015.
[3] 李颖. 间充质干细胞条件培养基对糖尿病肾病大鼠肾脏的保护作用[J]. 中华老年多器官疾病杂志, 2017, 2(16):140-143.
[4] Wang M T, Lin S C, Tang P L, et al. The impact of DPP-4 inhibitors on long-term survival among diabetic patients after first acute myocardial infarction[J]. Cardiovasc Diabetol, 2017, 16(1):89.
[5] Cordero O J, Varela-Calvino R, Lopez-Gonzalez T, et al. Anti-CD26 autoantibodies are involved in rheumatoid arthritis and show potential clinical interest[J]. Clin Biochem, 2017, 50(16-17):903-910.
[6] Fernandez-Paredes L, Casrouge A, Decalf J, et al. Multimarker risk stratification approach at multiple sclerosis onset[J]. Clin Immunol, 2017, 181:43-50.
[7] Papazafiropoulou A K, Papanas N, Trikkalinou A, et al. The oral dipeptidyl-peptidase-4 inhibitor sitagliptin Increases circulating levels of stromal-derived factor-1 alpha[J]. Exp Clin Endocrinol Diabetes, 2018, 126(6):367-370.
[8] Farag S S, Nelson R, Cairo M S, et al. High-dose sitagliptin for systemic inhibition of dipeptidylpeptidase-4 to enhance engraftment of single cord umbilical cord blood transplantation[J]. Oncotarget, 2017, 8(66):110350-110357.
[9] Waget A, Cabou C, Masseboeuf M, et al. Physiological and pharmacological mechanisms through which the DPP-4 inhibitor sitagliptin regulates glycemia in mice[J]. Endocrinology, 2011, 152(8):3018-3029.
[10] 刘珊珊. LC-MS/MS法分析人血浆中西格列汀浓度及其应用研究[J]. 天津医药, 2018, 10(46):1096-1101.
[11] Balakrishnan A, Gloude N, Sasik R, et al. Proinflammatory dual receptor T cells in chronic graft-versus-host disease[J]. Biol Blood Marrow Transplant, 2017, 23(11):1852-1860.
[12] 范一帆,齐丹,刘佳馨,等.鞘磷脂合酶抑制剂对血管紧张素Ⅱ诱导的高血压小鼠心肌纤维化的影响[J].首都医科大学学报,2016,37(6):731-735.

[1]	陈峰, 郑晓红, 李文斌. 脑胶质瘤免疫治疗进展[J]. 首都医科大学学报, 2019, 40(6): 966-971.
[2]	张亚敏, 王瑞云, 李军, 陶娟, 涂亚庭. B细胞活化因子的双向性:促进调节性T细胞分泌白细胞介素-35[J]. 首都医科大学学报, 2017, 38(5): 640-644.
[3]	宋冰冰, 陆小凡, 翁文佳, 粟斌, 张彤, 高艳青. HIV-1慢性感染者外周血中CD8⁺干细胞样记忆型T细胞变化以及对疾病进展的影响[J]. 首都医科大学学报, 2017, 38(5): 650-653.
[4]	郭轶先, 张蓝方, 孙雪静, 万岁桂, 夏长青. 紫外线照射未成熟树突状细胞诱导小鼠同种抗原免疫耐受及其机制研究[J]. 首都医科大学学报, 2017, 38(3): 431-438.
[5]	孙海青, 娄金丽, 刘新, 党燕, 时红波, 陈煜. 急性肝衰竭时外周血和肝组织中调节性T细胞表达变化的初探[J]. 首都医科大学学报, 2014, 35(4): 447-451.
[6]	张晓慧, 刘新, 白丽, 陈煜, 段钟平. 肝脏CD4+CD25+Foxp3+调节性T细胞在小鼠急、慢性肝损伤中的表达差异及意义[J]. 首都医科大学学报, 2014, 35(4): 483-487.
[7]	刘艳, 万岁桂. 滤泡辅助性T细胞与自身免疫性疾病的研究进展[J]. 首都医科大学学报, 2014, 35(4): 507-510.
[8]	李珍, 吴昊, 张彤. 男男同性恋人群HIV-1早期感染者T细胞活化水平分析[J]. 首都医科大学学报, 2014, 35(1): 92-95.
[9]	刘新, 娄金丽, 白丽, 张晓慧, 丁美, 段钟平. 流式细胞术分析小鼠肝纤维化时机体免疫细胞的变化[J]. 首都医科大学学报, 2013, 34(4): 540-544.
[10]	娄玮;王成硕;张罗. 调节性T细胞在过敏性鼻炎及抗原特异性免疫治疗中的作用[J]. 首都医科大学学报, 2012, 33(6): 729-734.
[11]	梁骑;焦艳梅;计云霞;李田;吴昊;张国元;唐中. 慢性丙型肝炎患者DNT细胞及T细胞亚群的研究[J]. 首都医科大学学报, 2012, 33(2): 214-217.
[12]	李娟;焦艳梅;高艳青;寇卜心;张彤;吴昊. Alu-LTR PCR方法检测HAART治疗系列各细胞亚群的整合型HIV-1 DNA[J]. 首都医科大学学报, 2011, 32(4): 449-452.
[13]	黄舒燕;吴昊;张明;孙欣;黄晓婕;焦艳梅;李娟;高艳青. 早期梅毒对HIV感染者CD4⁺T细胞计数的影响[J]. 首都医科大学学报, 2011, 32(4): 453-455.
[14]	焦艳梅;福军亮;张政;计云霞;王蕊;张彤;陈德喜;吴昊. HIV感染者调节性T细胞消耗可导致HIV特异CD8⁺细胞凋亡[J]. 首都医科大学学报, 2011, 32(2): 186-189.
[15]	画伟;焦艳梅;田亚坤;李娟;计云霞;王蕊;张彤;陈德喜;吴昊 . 调节性CD4⁺T细胞及非调节性CD4⁺T细胞与HIV/AIDS患者疾病进展关系的研究[J]. 首都医科大学学报, 2010, 31(6): 706-710.

通过渗透微泵稳定抑制小鼠血浆CD26/DPPIV分子酶活性的研究

Investigation on the inhibition of Sitagliptin by osmotic pump on CD26/DPPIV activity

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价