丹参注射液诱导类风湿关节炎成纤维样滑膜细胞NF-κB表达研究

中国临床药理学与治疗学 ›› 2012, Vol. 17 ›› Issue (8): 846-850.

丹参注射液诱导类风湿关节炎成纤维样滑膜细胞NF-κB表达研究

刘青松^1,2,3, 朱兴春², 蒋红^1,3, 蔡燕^1,2,3, 袁国华^1,3, 杨明辉^1,2,3, 张均², 李君安², 唐中^1,2,3

¹川北医学院附属医院风湿免疫研究所,²检验科,³风湿与感染免疫实验室,南充 637000,四川

收稿日期:2012-02-28 修回日期:2012-07-11 出版日期:2012-08-26 发布日期:2012-08-14
通讯作者: 唐中,教授、主任技师,硕士生导师,主要从事结缔组织病症的临床与基础研究。E-mail: tz5331@163.com
作者简介:刘青松,硕士,讲师,从事风湿病的临床与实验研究。E-mail: liuqingsong00@yahoo.com.cn
基金资助:
四川省卫生厅科研项目(090141,100151);川北医学院重点实验室开放基金[KFJJ(09)-04]

The changes of NF-κB promote rheumatoid arthritis fibroblast-like synoviocytes apoptosis by Danshen injection

LIU Qing-song^1,2,3, ZHU Xing-chun², JIANG Hong^1,3, CAI Yan^1,2,3, YUAN Guo-hua^1,3, YANG Ming-hui^1,2,3, ZHANG Jun², LI Jun-an², TANG Zhong^1,2,3

¹Institute of Rheumatology and Immunology, ²Department of Clinical Laboratory, ³Laboratory of Rheumatism and Infection Immunity, Affiliated Hospital Northern Sichuan Medical College, Nanchong 637000, Sichuan, China

Received:2012-02-28 Revised:2012-07-11 Online:2012-08-26 Published:2012-08-14

摘要/Abstract

摘要： 目的: 探讨丹参注射液对类风湿关节炎(RA)患者成纤维样滑膜细胞(FLSs)P65和P50表达及其在细胞内定位的影响。方法: RA患者关节液原代培养获取FLSs,终浓度为0、0.2 和 0.4 mg/mL 的丹参注射液作用 24 h 后,采用细胞免疫化学方法对P65和P50表达定位进行分析;提取核蛋白,采用免疫印迹方法检测P65和P50表达。结果: 细胞免疫化学结果显示,在无药物刺激的条件下,P65和P50均主要集中在细胞质中;当经 0.2 mg/mL 丹参注射液作用 24 h,P65和P50均有向细胞核转移趋势;当丹参注射液浓度为 0.4 mg/mL 时,P65和P50在细胞核中有明显聚集。免疫印迹结果表明,P65和P50在细胞核中表达量随丹参注射液浓度升高而增加,并当丹参注射液浓度为 0.4 mg/mL 时,二者的表达相比药物作用前显著增加,具有统计学意义(P<0.05)。结论: 丹参注射液可调节RA FLSs P65和P50从细胞质向细胞核转移。

关键词: 丹参, 类风湿关节炎, 成纤维样滑膜细胞, NF-κB

Abstract: AIM: To investigate the changes of expressions and the location of P65 and P50 in rheumatoid arthritis fibroblast-like synoviocytes (FLSs) by Danshen injection.METHODS: Fibroblast-like synovial cells obtained from synovial fluid by primary culture, and then treated with final concentrations of 0, 0.2 and 0.4 mg/mL of Danshen injection for 24 h. The expressions and location of P65 and P50 were detected with immunocytochemistry and Western blot.RESULTS: Immunocytochemistry showed that in the absence of drugs under the conditions of stimulation, either P65 or P50 was mainly concentrated in the cytoplasm, when the final concentration of Danshen injection reaches 0.2 mg/mL, both of them had a tendency to nuclear transfer, when the final concentration of Danshen injection to 0.4 mg/mL, both of them significantly accumulated in the nucleus. Western blot results showed that the expressions of P65 and P50 in the nucleus increased with the elevated concentration of Danshen injection, and when the concentration of Danshen injection reach to 0.4 mg/mL, the expressions of both of them increased significantly (P<0.05, n=4).CONCLUSION: Danshen injection can adjust rheumatoid arthritis fibroblast-like synoviocytes P65 and P50 from the cytoplasm to the cell nuclear transfer, and thus promote apoptosis of fibroblast-like synoviocytes.

Key words: Danshen injection, Rheumatoid arthritis, Fibroblast-like synoviocytes, NF-κB

中图分类号:

R593.22

刘青松, 朱兴春, 蒋红, 蔡燕, 袁国华, 杨明辉, 张均, 李君安, 唐中. 丹参注射液诱导类风湿关节炎成纤维样滑膜细胞NF-κB表达研究[J]. 中国临床药理学与治疗学, 2012, 17(8): 846-850.

LIU Qing-song, ZHU Xing-chun, JIANG Hong, CAI Yan, YUAN Guo-hua, YANG Ming-hui, ZHANG Jun, LI Jun-an, TANG Zhong. The changes of NF-κB promote rheumatoid arthritis fibroblast-like synoviocytes apoptosis by Danshen injection[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(8): 846-850.

参考文献

[1] Muller LU, Pap T. Pathogenesis of RA: more than just immune cells[J]. J Rheumatol, 2005, 64(6): 396-401.
[2] 周凌燕, 胡筱薇. 丹参注射液辅助治疗类风湿关节炎疗效观察[J]. 现代中西医结合杂志, 2011, 20(16): 1960-1961, 1987.
[3] 沈鹰, 刘正民, 黄清春, 等. 中西医结合治疗类风湿性关节炎27例[J]. 中国中西医结合杂志, 1994, 14(10):630.
[4] Wang X, Morris-Natschke SL, Lee KH. New developments in the chemistry and biology of the bioactive constituents of Tanshen[J]. Med Res Rev, 2007, 27(1):133-148.
[5] 蒋红, 刘青松, 凡瞿明, 等. 丹参体外抑制类风湿关节炎成纤维样滑膜细胞增殖[J]. 中华临床医师杂志:电子版, 2010, 4(4): 425-429.
[6] 刘青松, 朱兴春, 蒋红, 等. 丹参诱导类风湿关节炎成纤维样滑膜细胞bcl-2、bax基因表达研究[J]. 现代中西医结合杂志, 2010, 19(21): 2620-2622.
[7] 刘青松, 凡瞿明, 蒋红, 等. 丹参诱导类风湿关节炎患者成纤维样滑膜细胞凋亡[J]. 西安交通大学学报:医学版, 2010, 31(6): 763-765
[8] 刘青松, 朱兴春, 邢艳, 等. 丹参通过上调Fas/FasL基因表达诱导类风湿关节炎患者成纤维样滑膜细胞凋亡[J]. 第三军医大学学报, 2010, 32(22): 2452-2454.
[9] 刘青松, 蒋红, 唐中, 等. 丹参对类风湿关节炎患者滑膜细胞肿瘤坏死因子α表达的影响[J]. 中华临床医师杂志:电子版, 2009, 3(8): 1354-1358.
[10] 杜冠华, 张均田. 丹参现代研究概况与进展(续一) [J]. 医药导报, 2004, 23(6) : 355-359.
[11] 曾彩雯, 夏春华, 熊玉卿. 丹参及其制剂主要效应组分的药代动力学研究进[J]. 中国临床药理学与治疗学, 2011, 16(2) : 237-240.
[12] 毕明慧, 陈坚. 丹参素诱导人肝癌细胞株SMMC7721凋亡的研究[J]. 胃肠病学, 2011, 16(4): 222-225.
[13] Hao Y, Xie T, Korotcov A, et al. Salvianolic acid B inhibits growth of head and neck squamous cell carcinoma in vitro and in vivo via cyclooxygenase-2 and apoptotic pathways[J]. Int J Cancer, 2009, 124(9): 2200-2209.
[14] 温雅, 毕蕾, 张义彪, 等. 丹参水提液对肺癌A549细胞增殖的抑制作用[J]. 中医药导报, 2010, 16(1): 2-5.
[15] Kucharczak J, Simmons MJ, Fan Y, et al. To be, or not to be: NF-κB is the answer - role of Rel/NF- κB in the regulation of apoptosis[J]. Oncogene, 2003, 22(56): 8961-8982.
[16] Mattson MP, Meffert MK. Roles for NF-kappa B in nerve cell survival, plasticity, and disease[J]. Cell Death Differ, 2006, 13: 852-860.
[17] Tao Y, Yan D, Yang Q, et al. Low K⁺ promotes NF-kappaB/DNA binding in neuronal apoptosis induced by K⁺ loss[J]. Mol Cell Biol, 2006, 26(3): 1038-1050.
[18] Kasibhatla S, Brunner T, Genestier L, et al. DNA damaging agents induce expression of Fas ligand and subsequent apoptosis in T lymphocytes via the activation of NF-kappa B and AP-1[J]. Mol Cell, 1998, 1(4): 543- 551.
[19] 张晓红, 苏立达, 吕庆华, 等. 三氧化二砷诱导K562 细胞凋亡过程中IJB-A, NF-JB蛋白表达的研究[J]. 实用肿瘤杂志, 2004, 19(4): 292-295.
[20] 苏剑东, 昊灵飞. NF-κB与细胞凋亡[J]. 世界华人消化杂志, 2007, 15(12): 1411-1416.
[21] Yamasaki S, Nakashima T, Kawakami A, et al. Functional changes in rheumatoid fibroblast-like synovial cells through activation of peroxisome proliferator-activated receptor γ-mediated signalling pathway[J]. Clin Exp Immunol, 2002, 129(2):379-384.
[22] Stadel D, Cristofanon S, Abhari BA, et al. Requirement of nuclear factor κB for Smac mimetic-mediated sensitization of pancreatic carcinoma cells for gemcitabine-induced apoptosis[J]. Neoplasia, 2011, 13(12): 1162-1170.