重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白抑制强直性脊柱炎趋化因子CXCL16及其受体CXCR6的表达

doi:10.6040/j.issn.1671-7554.0.2015.253

山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (12): 51-56.doi: 10.6040/j.issn.1671-7554.0.2015.253

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白抑制强直性脊柱炎趋化因子CXCL16及其受体CXCR6的表达

张培艺¹, 周淑芬², 王梅英², 雷振华², 翁俊雄², 田野², 彭鑫², 傅碧玲², 邓柳霞², 邹世海², 郭成山^1,2

1. 山东大学第二医院血液科, 山东济南 250033;
2. 南方医科大学附属深圳宝安医院风湿免疫科, 广东深圳 518101

收稿日期:2015-03-09 出版日期:2015-12-10 发布日期:2015-12-10
通讯作者: 郭成山。E-mail:guochengshan1@163.com E-mail:guochengshan1@163.com
基金资助:
国家自然科学基金(81270577)

Expression of chemokine CXCL16 and its receptor CXCR6 can be suppressed by rcombinant human TNF receptor α Ⅱ-Ig fusion protein in ankylosing spondylitis

ZHANG Peiyi¹, ZHOU Shufen², WANG Meiying², LEI Zhenhua², WENG Junxiong², TIAN Ye², PENG Xin², FU Biling², DENG Liuxia², ZOU Shihai², GUO Chengshan^1,2

1. Department of Hematology, Second Hospital of Shandong University, Jinan 250033, Shandong, China;
2. Department of Rheumatology, Shenzhen Baoan Hospital Affiliated to Southern Medical University, Shenzhen 518101, Guangdong, China

Received:2015-03-09 Online:2015-12-10 Published:2015-12-10

摘要/Abstract

摘要： 目的探讨趋化因子CXCL16及其受体CXCR6(CXCL16/CXCR6)在强直性脊柱炎(AS)发病中的作用机制,及重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)的作用机制。方法实验分为AS活动组、AS治疗组及健康对照组,用酶联免疫吸附试验(ELISA)和荧光定量PCR法分别检测血清细胞核因子-κB受体活化因子配体(RANKL)、骨保护素(OPG)、CXCLl6水平及外周血单个核细胞(PBMC)CXCL16、CXCR6 mRNA表达水平;采用CCK-8法和ELISA法分别检测不同浓度CXCLl6重组蛋白(40、80 ng/mL)刺激淋巴细胞的增殖及培养上清中RANKL的表达水平。结果 ①AS活动组经rhTNFR:Fc治疗后,临床效果达95.45%,且AS治疗组各疗效指标血沉(ESR)、C反应蛋白(CRP)、Bath AS活动指数(BASDAI)、Bath AS功能指数(BASFI)、脊柱痛、夜间痛均较AS活动组下降(P均<0.05);② AS活动组RANK水平、RANKL/OPG比值均高于健康对照组(P均<0.05); AS活动组血清CXCL16的表达及PBMC之CXCL16、CXCR6 mRNA的表达均高于健康对照组及AS治疗组(P均<0.05);③在CXCLl6(40、80 ng/mL)刺激下,AS活动组淋巴细胞增殖能力高于无CXCLl6组(P均<0.05),且培养上清中RANKL表达水平高于无CXC Ll6组(P均<0.05);④ AS活动组血清CXCL16与RANKL、RANKL/OPG比值及ESR、CRP等实验室指标呈正相关(P均<0.05)。结论 CXCL16/CXCR6在AS的发病中可能发挥着重要作用,且rhTNFR:Fc降低CXCL16/CXCR6的表达可能是其抑制AS炎症反应及骨质破坏的重要作用机制之一。

关键词: 强直性脊柱炎, 趋化因子受体CXCR6, 核因子-&kappa, 趋化因子CXCL16, B受体活化因子配体, 骨保护素

Abstract: Objective To investigate the role of CXCL16/CXCR6 in the pathogenesis of ankylosing spondylitis (AS) and the functional mechanism of recombinant human TNF receptor-αⅡ-Ig fusion protein (rhTNFR:Fc). Methods The subjects were divided into AS active group, AS treated group and healthy control group. The levels of receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG) and CXCL16 in serum and CXCL16, CXCR6 mRNA in peripheral blood cells (PBMC) were detected with enzyme-linked immunosorbent assay (ELISA) and fluorescent quatitative PCR. The proliferation of lymphocytes and the level of RANKL in supernatant stimulated by different concentrations (40 ng/mL, 80 ng/mL) of rhCXCL16 were assessed with CCK-8 and ELISA. Results ① The clinical response rate in the AS active group reached 95.45%, and indicators including ESR, CRP, BASDAI, BASFI, spinal pain and nocturnal pain in AS treated group were significantly reduced compared with those in AS active group (all P<0.05). ② Serum RANKL level, RANKL/OPG ratio, and CXCLl6 level in AS active group were significantly higher than those in the healthy control group; CXCLl6 and CXCR6 mRNA levels in AS active group were both significantly higher than those in healthy control group and AS treated group (all P<0.05). ③ In the AS active group, stimulated by CXCLl6(40 ng/mL, 80 ng/mlL), the proliferation of lymphocytes and the level of RANKL in the supernatant were significantly higher (all P<0.05). ④ In the AS active group, the serum CXCL16 was positively correlated to RANKL, RANKLL/OPG radio, CRP and ESR (all P<0.05). Conclusion CXCL16/CXCR6 may play an important role in the pathogenesis of AS.Besides, rhTNFR:Fc could reduce the expression of CXCL16/CXCR6, which may be one of the mechanisms in inhibiting inflammation and bone destruction.

Key words: Osteoprotegerin, Ankylosing spondylitis, Chemokine CXCL16, Chemokine receptor CXCR6, Receptor activator of nuclear factor κB-ligand

中图分类号:

R593.23

张培艺, 周淑芬, 王梅英, 雷振华, 翁俊雄, 田野, 彭鑫, 傅碧玲, 邓柳霞, 邹世海, 郭成山. 重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白抑制强直性脊柱炎趋化因子CXCL16及其受体CXCR6的表达[J]. 山东大学学报（医学版）, 2015, 53(12): 51-56.

ZHANG Peiyi, ZHOU Shufen, WANG Meiying, LEI Zhenhua, WENG Junxiong, TIAN Ye, PENG Xin, FU Biling, DENG Liuxia, ZOU Shihai, GUO Chengshan. Expression of chemokine CXCL16 and its receptor CXCR6 can be suppressed by rcombinant human TNF receptor α Ⅱ-Ig fusion protein in ankylosing spondylitis[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2015, 53(12): 51-56.

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重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白抑制强直性脊柱炎趋化因子CXCL16及其受体CXCR6的表达

Expression of chemokine CXCL16 and its receptor CXCR6 can be suppressed by rcombinant human TNF receptor α Ⅱ-Ig fusion protein in ankylosing spondylitis

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