A771726对体外培养胶原性关节炎大鼠腹腔巨噬细胞产生一氧化氮的影响

中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (9): 965-970.

A771726对体外培养胶原性关节炎大鼠腹腔巨噬细胞产生一氧化氮的影响

王婷玉^1,2, 李俊², 金芝贵¹, 吴飞华¹, 王小华², 周倩²

¹上海交通大学医学院附属第九人民医院药剂科,上海 200011;
²安徽医科大学药学院,合肥 230032,安徽

收稿日期:2011-08-16 修回日期:2011-09-09 出版日期:2011-09-26 发布日期:2020-09-16
通讯作者: 李俊,男,教授,博士生导师,研究方向:抗炎免疫药理学、临床药理学。Tel: 0551-5161001 E-mail: aydlijun@sohu.com
作者简介:王婷玉,女,博士,博士后,药师,研究方向:抗炎免疫药理学、临床药学。Tel: 021-23271699-5368 E-mail: wangty1982@gmail.com

In vitro effect of A771726 on the secretion of nitric oxide of peritoneal macrophages from collagen-induced arthritis rats

WANG Ting-yu^1,2, LI Jun², JIN Zhi-gui¹, WU Fei-hua¹, WANG Xiao-hua², ZHOU Qian²

¹Department of Pharmacy, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China;
²School of Pharmacy, Anhui Medical University, Hefei 230032, Anhui, China

Received:2011-08-16 Revised:2011-09-09 Online:2011-09-26 Published:2020-09-16

摘要/Abstract

摘要： 目的: 在体外培养环境中,研究来氟米特活性代谢产物A771726,对脂多糖(LPS)刺激的胶原性关节炎(collagen-induced arthritis, CIA)大鼠腹腔巨噬细胞(peritoneal macrophages, PMФ)产生一氧化氮(nitric oxide, NO)的影响及作用机制。方法: 建立大鼠CIA模型,无菌制备CIA大鼠PMФ悬液,A771726体外给药,硝酸还原法测定A771726 影响CIA大鼠PMФ分泌NO的量效和时效关系;MTT法检测A771726对CIA大鼠PMФ细胞活力的影响;RT-PCR法测定A771726对CIA大鼠PMФ中诱导型一氧化氮合酶(inducible NO synthase, iNOS) mRNA表达的影响;Western blot法检测A771726对CIA大鼠PMФ中iNOS蛋白表达的影响。结果: A771726体外用药,可有效降低CIA大鼠PMФ中NO的分泌,且抑制作用有明显的量效和时效关系;A771726在有效抑制NO分泌的浓度范围和作用时间内,对PMФ的细胞活力无明显作用;A771726 (0.1、1、10 μmol/L)能显著抑制CIA大鼠PMФ iNOS mRNA和iNOS蛋白的表达。结论: A771726 (0.1、1、10 μmol/L)可能通过抑制CIA大鼠PMФ iNOS mRNA的转录,进而影响iNOS蛋白的翻译,最终引起NO的分泌减少。该作用并非由A771726对PMФ的毒性引起。

关键词: A771726, 关节炎, 巨噬细胞, 诱导型一氧化氮合酶

Abstract: AIM: To study the effect and mechanism of A771726, the active metabolite of leflunomide, on the production of nitric oxide (NO) of peritoneal macrophages (PMФ) from collagen-induced arthritis (CIA) rats. METHODS: Collagen-induced arthritis (CIA) model was induced in rats and the PMФ were collected. A771726 were used in vitro. The dose-dependent and time-dependent effects of A771726 on the secretion of NO of PMФ from CIA rats were checked by nitric acid reduction synthase method; MTT reagent was used to detect the effect of A771726 on the cell viability of PMФ from CIA rats; RT-PCR was used to detect the expression of inducible NO synthase (iNOS) mRNA. Western blot was used to detect the expression of iNOS protein. RESULTS: A771726 could inhibit the secretion of NO of PMФ from CIA rats in a dose-dependent and time-dependent manner. The cytotoxicity of A771726 was not detected during the time course and dose range. A771726 (0.1, 1, 10 mol/L) significantly inhibited the expression of iNOS mRNA and iNOS protein in PMФ from CIA rats. CONCLUSION: A771726 (0.1, 1, 10 mol/L) can inhibit the transcription of iNOS mRNA in PMФ from CIA rats, and reduce the translation of iNOS protein, which leads to suppressing the secretion of NO. And the suppression is not related to the cytotoxicity of A771726.

Key words: A771726, Arthritis, Macrophages, iNOS

中图分类号:

R965.2

王婷玉, 李俊, 金芝贵, 吴飞华, 王小华, 周倩. A771726对体外培养胶原性关节炎大鼠腹腔巨噬细胞产生一氧化氮的影响[J]. 中国临床药理学与治疗学, 2011, 16(9): 965-970.

WANG Ting-yu, LI Jun, JIN Zhi-gui, WU Fei-hua, WANG Xiao-hua, ZHOU Qian. In vitro effect of A771726 on the secretion of nitric oxide of peritoneal macrophages from collagen-induced arthritis rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2011, 16(9): 965-970.

参考文献

[1] Herrmann ML, Schleyerbach R, Kirschbaum BJ. Leflunomide: an immunomodulatory drug for the treatment of rheumatoid arthritis and other autoimmune diseases [J]. Immunopharmacology, 2000, 47(2/3): 273-289.
[2] Burger D, Begue-Pastor N, Benavent S, et al. The active metabolite of leflunomide, A771726, inhibits the production of prostaglandin E(2), matrix metalloproteinase 1 and interleukin 6 in human fibroblast-like synoviocytes [J]. Rheumatology(Oxford), 2003, 42(1): 89-96.
[3] 王小华, 李俊, 王婷玉, 等. A771726体外给药对LPS刺激的胶原性关节炎大鼠腹腔巨噬细胞免疫功能的影响[J]. 安徽医科大学学报, 2009, 44(3): 362-365.
[4] 胡国伟, 董群. 大蒜素对白色念珠菌感染的免疫抑制小鼠巨噬细胞功能的影响[J]. 中国临床药理学与治疗学, 2011, 16(3): 270-273.
[5] Plüddemann A, Mukhopadhyay S, Gordon S. Innate immunity to intracellular pathogens: macrophage receptors and responses to microbial entry [J]. Immunol Rev, 2011, 240(1):11-24.
[6] Harford KA, Reynolds CM, McGillicuddy FC, et al. Fats, inflammation and insulin resistance: insights to the role of macrophage and T-cell accumulation in adipose tissue [J]. Proc Nutr Soc, 2011, 12: 1-10.
[7] Ma Y, Wang X, Wu X, et al.(Z)-5-(4-methoxybenzylidene) thiazolidine-2, 4-dione ameliorates the adjuvant-induced arthritis via inhibiting the migration of macrophage and down-regulating the cytokine mRNA expression [J]. Int Immunopharmacol, 2010, 10(11): 1456-1462.
[8] Bogdan C, Rollinghoff M, Diefenbach A. The role of nitric oxide in innate immunity [J]. Immunol Rev, 2000, 173: 17-26.
[9] 茅尧生, 曹淼英, 周鑫. 强化胰岛素治疗对脓毒症患者血清血管性假血友病因子、内皮素1和一氧化氮浓度及预后的影响[J]. 中国临床药理学与治疗学, 2010, 15(12): 1414-1417.
[10] Cook H, Cattell V. Role of nitric oxide in immune-mediated diseases [J]. Clin Sci, 1996, 91(4): 375-384.
[11] Wang JX, Hou LF, Yang Y, et al. SM905, an artemisinin derivative, inhibited NO and pro- inflammatory cytokine produced by suppressiong MAPK and NF-κB pathways in RAW 264.7 macrophages [J]. Acta Pharmacol Sin, 2009, 30(10): 1428-1435.

[1]	廖梦玲, 汪艳, 罗静, 王诺妍, 华玲, 张瑜, 邓非, 袁月, 周军, 周红. 青蒿琥酯抑制巨噬细胞释放前炎症细胞因子的分子作用机制研究[J]. 中国临床药理学与治疗学, 2023, 28(9): 969-978.
[2]	范杰, 金永明, 江孝龙, 蒋国华. lncRNA HOTAIR调控miR-206影响类风湿关节炎滑膜细胞增殖和凋亡的分子机制研究[J]. 中国临床药理学与治疗学, 2023, 28(7): 736-742.
[3]	王鑫昱, 陈亦鹏, 马永力, 何君妃. “杜仲-当归”调控NLPR3炎症小体对骨关节炎大鼠改善作用的研究[J]. 中国临床药理学与治疗学, 2023, 28(7): 751-757.
[4]	王铁烽, 苏洁, 王伟东, 夏荣双, 李思凡, 朱文瑞, 楼招欢. 三叶青调节Th17/Treg平衡抗类风湿性关节炎的作用及机制研究[J]. 中国临床药理学与治疗学, 2023, 28(2): 138-146.
[5]	王亮, 张虎林, 汪小敏, 杨超强, 王宜灿, 赖学倩. 铁超载在骨关节炎发生发展中的作用机制研究进展[J]. 中国临床药理学与治疗学, 2022, 27(9): 1075-1080.
[6]	潘淑, 吴义金, 张洒洒, 王启海, 罗婷婷, 殷勤. 基于OAT3介导的MTX治疗佐剂诱导性关节炎大鼠的药动学研究[J]. 中国临床药理学与治疗学, 2022, 27(5): 516-525.
[7]	周世琴, 骆亚莉, 周雯, 齐晓风, 肖孟勇. 肺纤维化的相关分子机制和治疗现状[J]. 中国临床药理学与治疗学, 2022, 27(10): 1133-1147.
[8]	易虹, 顾兵, 陈玉年, 李华南. 中性粒细胞胞外诱捕网及其在痛风性疾病中的作用[J]. 中国临床药理学与治疗学, 2021, 26(10): 1167-1173.
[9]	孔祥, 华强, 姚新明, 孟祥健, 钟民, 郭莉群. 芝麻酚调控巨噬细胞极化改善肥胖小鼠脂肪组织炎症及胰岛素抵抗[J]. 中国临床药理学与治疗学, 2020, 25(7): 728-733.
[10]	邰宇, 张子璇, 张玲玲. 银屑病关节炎治疗进展[J]. 中国临床药理学与治疗学, 2020, 25(12): 1395-1407.
[11]	胡培培，黄芳. 云南白药通过抗炎作用缓解碘乙酸钠诱导的大鼠膝骨关节炎疼痛[J]. 中国临床药理学与治疗学, 2019, 24(3): 254-259.
[12]	程润，丁文溪，严尚学，魏伟. 人脐带来源间充质干细胞对佐剂性关节炎大鼠巨噬细胞功能的影响[J]. 中国临床药理学与治疗学, 2019, 24(12): 1335-1340.
[13]	邹丽霞，卢美萍，徐益萍，郑琪，郑嵘君. 托珠单抗与依那西普治疗多关节炎型幼年特发性关节炎临床研究[J]. 中国临床药理学与治疗学, 2019, 24(12): 1421-1427.
[14]	曹迎亚，陈群，王箴，吴敬医，姜小敢，金孝岠，鲁卫华. 巨噬细胞在脂多糖诱导肠上皮细胞凋亡中的作用[J]. 中国临床药理学与治疗学, 2019, 24(10): 1142-1146.
[15]	韩晨阳，杨毅，李文燕，王瑾，郭丽. MicroRNA-136靶向CD163抑制CD68⁺CD163⁺M2型巨噬细胞极化的作用研究[J]. 中国临床药理学与治疗学, 2019, 24(1): 1-8.