论文部分内容阅读
摘 要 目的:考察甘草附子汤通过抗滑膜血管生成途径对佐剂性关节炎(AA)模型小鼠的治疗作用机制。方法:取雄性Balb/c小鼠48只随机分为正常组、模型组、甘草附子汤组和雷公藤多苷组(阳性药物组),每组12只。除正常组外,其余小鼠左后足趾皮内一次性注射弗氏完全佐剂以建立AA模型。造模12 d后,正常组、模型组小鼠灌胃相应体积的水,甘草附子汤组(7.8 g/kg,以总生药量计算)和雷公藤多苷组(0.01 g/kg)小鼠灌胃相应药液20 mL/kg,每天1次,连续18 d。观察并记录小鼠关节病变情况,并采用水容积法测量小鼠足肿胀度;采用苏木精-伊红染色法观察小鼠踝关节组织病理学变化;采用免疫荧光法检测小鼠踝关节滑膜组织中血小板-内皮细胞黏附分子(CD31)、血管内皮生长因子(VEGF)的蛋白表达水平;采用Western blotting法检测小鼠踝关节滑膜组织中核转录因子κB(NF-κB)、锌指转录因子GATA4(GATA4)的蛋白表达水平。结果:与正常组比较,模型组小鼠足跖明显红肿,足肿胀度显著升高(P<0.05);踝关节滑膜组织增生,血管翳明显增多,可见大量炎症细胞及关节侵蚀等;滑膜组织中CD31、VEGF、NF-κB、GATA4的蛋白表达水平均显著升高(P<0.05)。与模型组比较,甘草附子汤组小鼠足跖红肿现象减轻,足肿胀度显著降低(P<0.05);踝关节滑膜组织中血管翳有所减少,其他病理症状有所改善;滑膜组织中CD31、VEGF、NF-κB、GATA4的蛋白表达水平均显著降低(P<0.05)。结论:甘草附子汤可通过下调关节滑膜组织中VEGF、NF-κB、GATA4蛋白的表达,减少滑膜组织中血管翳的形成并有效抑制血管新生,从而对AA模型小鼠发挥防治骨质破坏、保护关节等作用。
关键词 甘草附子汤;类风湿关节炎;佐剂性关节炎;血管新生;血小板-內皮细胞黏附分子;血管内皮生长因子;锌指转录因子GATA4;核因子κB;小鼠
Study on Anti-synovial Angiogenesis Mechanism of Licorice and Aconite Decoction in the Treatment of Adjuvant Arthritis Model Mice
CAI Yue1,LIU Xiaolong2,GUO Jing3,ZHANG Nan4(1. Dept. of Rheumatology, the First Affiliated Hospital of Tianjin University of TCM, Tianjin 300193, China; 2. Dept. of Hepatobilliary Surgery, Tianjin First Central Hospital, Tianjin 300192, China; 3. Dept. of Pharmacy, the First Affiliated Hospital of Tianjin University of TCM, Tianjin 300193, China; 4. School of Pharmacy, China Medical University, Shenyang 110003, China)
ABSTRACT OBJECTIVE: To investigate the therapeutic effect of Licorice and aconite decoction in the treatment of adjuvant arthritis (AA) model mice through anti-synovial angiogenesis pathway. METHODS: Totally 48 male Balb/c mice were randomly divided into normal group, model group, Licorice and aconite decoction group and tripterygium glycosides group (positive drug group), with 12 mice in each group. Except for normal group, AA model was established by intradermal injection of Freund’s complete adjuvant into the left hind toe of mice. 12 d after modeling, normal group and model group were given same volume of water intragastrically; Licorice and aconite decoction group (7.8 g/kg,by total amount of crude drug) and tripterygium glycosides group (0.01 g/kg) were given relevant medicine 20 mL/kg intragastrically, once a day, for consecutive 18 d. The joint lesions of mice were observed and recorded, and the foot swelling degree of mice was measured by water volume method. HE staining was used to observe the pathological change of ankle joint in mice. The protein levels of platelet endothelial cell adhesion molecule-1 (CD31) and vascular endothelial growth factor (VEGF) were determined by immunofluorescence assay. The protein expressions of nuclear factor κB(NF-κB) and zinc finger transcription factor GATA4 (GATA4) were detected by Western blotting assay. RESULTS: Compared with normal group, ankle joint of model group mice was markedly reddened and swollen, and foot swelling degree increased significantly (P<0.05). Synovial tissue of ankle joint proliferated, pannus increased significantly, and a large number of inflammatory cells and joint erosion were observed. The protein expression of CD31, VEGF, NF-κB and GATA4 in synovial tissue of mice were increased significantly (P<0.05). Compared with model group, the redness and swelling of ankle joint in mice were alleviated, and the foot swelling degree was significantly reduced in Licorice and aconite decoction group (P<0.05). Pannus in synovial tissue decreased and other pathological symptoms were improved. The protein expression of CD31, VEGF, NF-κB and GATA4 were decreased significantly in synovial tissue (P<0.05). CONCLUSIONS: Licorice and aconite decoction can decrease the protein expression of VEGF,NF-κB and GATA4 in synovial tissue, reduce the generation of pannus in synovial tissue and effectively inhibit the angiogenesis in synovial tissue so as to prevent bone destruction and protect joint of AA model mice. KEYWORDS Licorice and aconite decoction; Rheumatoid arthritis; Adjuvant arthritis; Angiogenesis; CD31; VEGF; GATA4; NF-κB; Mice
类风湿性关节炎(Rheumatoid arthritis,RA)是一种以慢性、进行性、侵袭性关节炎为主要表现的自身免疫性疾病;其全球患病率为0.5%~1%,临床表现为对称发作的关节肿胀、疼痛和持续晨僵等;其病理特征包括持续性滑膜炎、滑膜增生异常、血管生成和血管翳形成增加等,当血管翳逐渐扩展到关节面和关节软骨时,则会诱导软骨和骨骼被逐渐破坏,最终导致关节畸形[1]。血管生成已被确定为RA疾病进展的重要标志,并且通常被认为是从急性炎症期到慢性炎症期的“转换”标志[2]。这是由于血管生成不仅可维持滑膜关节内细胞群的扩张,而且还可进一步增强白细胞向组织中募集,导致炎症加重;而炎症状态的滑膜细胞会产生促血管生成因子,进一步诱导血管生成[3]。因此,抑制血管生成可能是RA临床治疗一种新思路[4]。
甘草附子汤收载于东汉张仲景的《伤寒论》,由甘草、附子、白术、桂枝组方而成,具有祛风除湿、温经散寒、温阳补中的功效,临床上用于治疗风湿性和类风湿性关节炎具有较好的效果[5]。弗氏佐剂关节炎(AA)模型是一种应用广泛的关节炎动物模型,常用于预测药物对RA的临床疗效[6]。本课题组前期研究发现,甘草附子汤能明显下调AA模型小鼠滑膜组织中基质金属蛋白酶1(MMP-1)和MMP-3的表达,降低其炎性细胞因子水平[7]。基于此,本研究拟进一步探讨甘草附子汤对AA模型小鼠滑膜组织中血管新生的影响,并通过检测滑膜组织中血小板-内皮细胞黏附分子(CD31)、血管内皮生长因子(VEGF)、锌指转录因子GATA4(GATA4)和核因子κB(NF-κB)的表达水平,探讨甘草附子汤对AA模型小鼠的治疗作用机制,以期为其用于临床治疗RA提供科学的理论依据。
1 材料
1.1 仪器
YLS-7B型足趾容积测量仪(北京吉安得尔科技有限公司);BX-51型荧光显微镜(日本Olympus公司);ESP-300型电泳仪(上海天能科技有限公司);Trans-Blot SD型槽式转膜仪、ChemiDoc MP型凝胶成像仪(美国Bio-Rad公司);SW-CJ-2D型超净工作台(无锡易纯净化设备有限公司);HQ96型分析天平(瑞士Mettler Toledo公司);RM2265型切片机(德国Leica公司)。
1.2 药品与试剂
甘草附子汤(组方药材炙甘草、炮附子、白术、桂枝饮片均购自天津同仁堂药店,经天津中医药大学张坚教授鉴定为真品;制备过程:炙甘草、炮附子、白术、桂枝各15 g,以1.2 L水煎煮1 h,滤过,减压浓缩,以水制成含总生药量约390 mg/mL的药液,4 ℃保存);雷公藤多苷片(江苏美通制药有限公司,批号:1211242063,规格:10 mg;使用时将其碾碎,以水制成含主药为0.5 mg/mL 的混悬液);鼠CD31单克隆抗体、兔VEGF多克隆抗体、兔NF-κB单克隆抗体、兔GATA4多克隆抗体、兔β-肌动蛋白(β-actin)多克隆抗体(英国Abcam公司,批号分别为ab119341、ab2349、ab109458、ab124265、ab8227);弗氏完全佐剂(美国Sigma公司,批号:100M8725);驴抗小鼠IgG(H+L)Alexa Fluor 488标记二抗、驴抗兔IgG(H+L)Alexa Fluor 488标记二抗(美国Thermo Fisher Scientific公司,批号分别为CA21202S、A-21206);山羊抗兔辣根过氧化物酶标记二抗(美国CST公司,批号:7074);4′,6-二脒基-2-苯基吲哚(DAPI)、马血清(美国Gibco公司);苏木精-伊红(HE)染料试剂盒、pH 7.4磷酸盐缓冲液(PBS)、Triton X-100(北京索莱宝科技有限公司);增强化学发光法(ECL)显色剂(美国Thermo Fisher Scientific公司);十二烷基磺酸钠-聚丙烯酰胺(SDS-PAGE)凝胶配制试剂盒、SDS-PAGE蛋白上样缓冲液、BCA蛋白浓度测定试剂盒(上海碧云天生物技术有限公司);水为无菌蒸馏水。
1.3 动物
8周龄Balb/c小鼠,雄性,体质量为(18±2)g,购自北京维通利华实验技术有限公司,动物生产合格证号SCXK(京)2014-0057,清洁级饲养。经天津中医药大学伦理委员会审查批准通过本次动物实验。
2 方法
2.1 分组、造模及给药
取小鼠48只,随机分为正常组、模型组、甘草附子汤组[7.8 g/kg(以总生药量计算);剂量根据成人临床日常用量,按照实验动物与人体表面积比换算]和雷公藤多苷组(阳性药物组,0.01 g/kg[7]),每组12只。除正常组注射等体积生理盐水外,其余组小鼠均于左后足趾皮内一次性注射弗氏完全佐剂50 μL以建立AA模型[8]。以小鼠造模侧、对侧和前爪足跖明显肿胀,受累关节畸形,行动不便且体质量明显下降判定为造模成功。造模12 d后,剔除造模失败的小鼠后,正常组、模型组小鼠灌胃水,甘草附子汤组和雷公藤多苷组小鼠灌胃相应药液,灌胃体积均为20 mL/kg,每天1次,连续18 d。
2.2 小鼠足肿胀情况观察
分别在造模前和给药结束时,观察并记录小鼠关节病变情況,同时采用水容积法测量小鼠右后足跖容积并计算足肿胀度[肿胀度(mL)=给药结束时足跖容积-造模前足跖容积]。
2.3 小鼠踝关节病理学检查
给药结束后,断颈处死小鼠,于无菌环境下取其右踝关节,石蜡包埋,切片(厚度:5 μm),脱水,HE染色,乙醇梯度脱水,二甲苯透明,中性树胶封片,在显微镜下观察踝关节病理学变化。 [13] 邓伟.甘草附子汤治疗膝骨关节炎的临床研究[J].中药材,2008,31(7):1107-1110.
[14] SUKE SG,NEGI H,MEDIRATTA PK,et al. Anti-arthri- tic and anti-inflammatory activity of combined pioglitazone and prednisolone on adjuvant-induced arthritis[J].Eur J Pharmacol,2013,718(1/2/3):57-62.
[15] ELSHABRAWY HA,CHEN Z,VOLIN MV,et al. The pathogenic role of angiogenesis in rheumatoid arthritis[J].Angiogenesis,2015,18(4):433-448.
[16] YANG X,QIU P,CHEN B,et al. KIAA1199 as a potential diagnostic biomarker of rheumatoid arthritis related to angiogenesis[J]. Arthritis Res Ther,2015. DOI:10.1186/s13075-015-0637-y.
[17] CAO D,HOU M,GUAN YS,et al. Expression of HIF-1α and VEGF in colorectal cancer:association with clinical outcomes and prognostic implications[J]. BMC Cancer,2009. DOI:10.1186/1471-2407-9-432.
[18] ZHANG J,LI C,ZHENG Y,et al. Inhibition of angiogenesis by arsenic trioxide via TSP-1-TGF-β1-CTGF-VEGF functional module in rheumatoid arthritis[J]. Oncotarget,2017,8(43):73529-73546.
[19] 李靜平,顾雯,倪艺榕,等.人参皂苷Rg3对糖尿病大鼠难愈创面表皮细胞及血管新生的影响[J].中国药理学通报,2019,35(4):551-556.
[20] MAIJER KI,NOORT AR,DE HAIR MJ,et al. Nuclear factor-κB-inducing kinase is expressed in synovial endothelial cells in patients with early arthritis and correlates with markers of inflammation:a prospective Cohort study[J]. J Rheumatol,2015,42(9):1573-1581.
[21] NOORT AR,VAN ZOEST KP,WEIJERS EM,et al.NF- kappaB-inducing kinase is a key regulator of inflammation-induced and tumour-associated angiogenesis[J]. J Pathol,2014,234(3):375-385.
[22] HEINEKE J,AUGER-MESSIER M,XU J,et al. Cardiomyocyte GATA4 functions as a stress-responsive regulator of angiogenesis in the murine heart[J]. J Clin Invest,2007,117(11):3198-3210.
[23] JIA W,WU W,YANG D,et al. GATA4 regulates angiogenesis and persistence of inflammation in rheumatoid arthritis[J]. Cell Death Dis,2018,9(5):503-518.
[24] SU CM,HUANG CY,TANG CH. Characteristics of resistin in rheumatoid arthritis angiogenesis[J]. Biomark Med,2016,10(6):651-660.
(收稿日期:2018-12-14 修回日期:2019-05-06)
(编辑:段思怡)
关键词 甘草附子汤;类风湿关节炎;佐剂性关节炎;血管新生;血小板-內皮细胞黏附分子;血管内皮生长因子;锌指转录因子GATA4;核因子κB;小鼠
Study on Anti-synovial Angiogenesis Mechanism of Licorice and Aconite Decoction in the Treatment of Adjuvant Arthritis Model Mice
CAI Yue1,LIU Xiaolong2,GUO Jing3,ZHANG Nan4(1. Dept. of Rheumatology, the First Affiliated Hospital of Tianjin University of TCM, Tianjin 300193, China; 2. Dept. of Hepatobilliary Surgery, Tianjin First Central Hospital, Tianjin 300192, China; 3. Dept. of Pharmacy, the First Affiliated Hospital of Tianjin University of TCM, Tianjin 300193, China; 4. School of Pharmacy, China Medical University, Shenyang 110003, China)
ABSTRACT OBJECTIVE: To investigate the therapeutic effect of Licorice and aconite decoction in the treatment of adjuvant arthritis (AA) model mice through anti-synovial angiogenesis pathway. METHODS: Totally 48 male Balb/c mice were randomly divided into normal group, model group, Licorice and aconite decoction group and tripterygium glycosides group (positive drug group), with 12 mice in each group. Except for normal group, AA model was established by intradermal injection of Freund’s complete adjuvant into the left hind toe of mice. 12 d after modeling, normal group and model group were given same volume of water intragastrically; Licorice and aconite decoction group (7.8 g/kg,by total amount of crude drug) and tripterygium glycosides group (0.01 g/kg) were given relevant medicine 20 mL/kg intragastrically, once a day, for consecutive 18 d. The joint lesions of mice were observed and recorded, and the foot swelling degree of mice was measured by water volume method. HE staining was used to observe the pathological change of ankle joint in mice. The protein levels of platelet endothelial cell adhesion molecule-1 (CD31) and vascular endothelial growth factor (VEGF) were determined by immunofluorescence assay. The protein expressions of nuclear factor κB(NF-κB) and zinc finger transcription factor GATA4 (GATA4) were detected by Western blotting assay. RESULTS: Compared with normal group, ankle joint of model group mice was markedly reddened and swollen, and foot swelling degree increased significantly (P<0.05). Synovial tissue of ankle joint proliferated, pannus increased significantly, and a large number of inflammatory cells and joint erosion were observed. The protein expression of CD31, VEGF, NF-κB and GATA4 in synovial tissue of mice were increased significantly (P<0.05). Compared with model group, the redness and swelling of ankle joint in mice were alleviated, and the foot swelling degree was significantly reduced in Licorice and aconite decoction group (P<0.05). Pannus in synovial tissue decreased and other pathological symptoms were improved. The protein expression of CD31, VEGF, NF-κB and GATA4 were decreased significantly in synovial tissue (P<0.05). CONCLUSIONS: Licorice and aconite decoction can decrease the protein expression of VEGF,NF-κB and GATA4 in synovial tissue, reduce the generation of pannus in synovial tissue and effectively inhibit the angiogenesis in synovial tissue so as to prevent bone destruction and protect joint of AA model mice. KEYWORDS Licorice and aconite decoction; Rheumatoid arthritis; Adjuvant arthritis; Angiogenesis; CD31; VEGF; GATA4; NF-κB; Mice
类风湿性关节炎(Rheumatoid arthritis,RA)是一种以慢性、进行性、侵袭性关节炎为主要表现的自身免疫性疾病;其全球患病率为0.5%~1%,临床表现为对称发作的关节肿胀、疼痛和持续晨僵等;其病理特征包括持续性滑膜炎、滑膜增生异常、血管生成和血管翳形成增加等,当血管翳逐渐扩展到关节面和关节软骨时,则会诱导软骨和骨骼被逐渐破坏,最终导致关节畸形[1]。血管生成已被确定为RA疾病进展的重要标志,并且通常被认为是从急性炎症期到慢性炎症期的“转换”标志[2]。这是由于血管生成不仅可维持滑膜关节内细胞群的扩张,而且还可进一步增强白细胞向组织中募集,导致炎症加重;而炎症状态的滑膜细胞会产生促血管生成因子,进一步诱导血管生成[3]。因此,抑制血管生成可能是RA临床治疗一种新思路[4]。
甘草附子汤收载于东汉张仲景的《伤寒论》,由甘草、附子、白术、桂枝组方而成,具有祛风除湿、温经散寒、温阳补中的功效,临床上用于治疗风湿性和类风湿性关节炎具有较好的效果[5]。弗氏佐剂关节炎(AA)模型是一种应用广泛的关节炎动物模型,常用于预测药物对RA的临床疗效[6]。本课题组前期研究发现,甘草附子汤能明显下调AA模型小鼠滑膜组织中基质金属蛋白酶1(MMP-1)和MMP-3的表达,降低其炎性细胞因子水平[7]。基于此,本研究拟进一步探讨甘草附子汤对AA模型小鼠滑膜组织中血管新生的影响,并通过检测滑膜组织中血小板-内皮细胞黏附分子(CD31)、血管内皮生长因子(VEGF)、锌指转录因子GATA4(GATA4)和核因子κB(NF-κB)的表达水平,探讨甘草附子汤对AA模型小鼠的治疗作用机制,以期为其用于临床治疗RA提供科学的理论依据。
1 材料
1.1 仪器
YLS-7B型足趾容积测量仪(北京吉安得尔科技有限公司);BX-51型荧光显微镜(日本Olympus公司);ESP-300型电泳仪(上海天能科技有限公司);Trans-Blot SD型槽式转膜仪、ChemiDoc MP型凝胶成像仪(美国Bio-Rad公司);SW-CJ-2D型超净工作台(无锡易纯净化设备有限公司);HQ96型分析天平(瑞士Mettler Toledo公司);RM2265型切片机(德国Leica公司)。
1.2 药品与试剂
甘草附子汤(组方药材炙甘草、炮附子、白术、桂枝饮片均购自天津同仁堂药店,经天津中医药大学张坚教授鉴定为真品;制备过程:炙甘草、炮附子、白术、桂枝各15 g,以1.2 L水煎煮1 h,滤过,减压浓缩,以水制成含总生药量约390 mg/mL的药液,4 ℃保存);雷公藤多苷片(江苏美通制药有限公司,批号:1211242063,规格:10 mg;使用时将其碾碎,以水制成含主药为0.5 mg/mL 的混悬液);鼠CD31单克隆抗体、兔VEGF多克隆抗体、兔NF-κB单克隆抗体、兔GATA4多克隆抗体、兔β-肌动蛋白(β-actin)多克隆抗体(英国Abcam公司,批号分别为ab119341、ab2349、ab109458、ab124265、ab8227);弗氏完全佐剂(美国Sigma公司,批号:100M8725);驴抗小鼠IgG(H+L)Alexa Fluor 488标记二抗、驴抗兔IgG(H+L)Alexa Fluor 488标记二抗(美国Thermo Fisher Scientific公司,批号分别为CA21202S、A-21206);山羊抗兔辣根过氧化物酶标记二抗(美国CST公司,批号:7074);4′,6-二脒基-2-苯基吲哚(DAPI)、马血清(美国Gibco公司);苏木精-伊红(HE)染料试剂盒、pH 7.4磷酸盐缓冲液(PBS)、Triton X-100(北京索莱宝科技有限公司);增强化学发光法(ECL)显色剂(美国Thermo Fisher Scientific公司);十二烷基磺酸钠-聚丙烯酰胺(SDS-PAGE)凝胶配制试剂盒、SDS-PAGE蛋白上样缓冲液、BCA蛋白浓度测定试剂盒(上海碧云天生物技术有限公司);水为无菌蒸馏水。
1.3 动物
8周龄Balb/c小鼠,雄性,体质量为(18±2)g,购自北京维通利华实验技术有限公司,动物生产合格证号SCXK(京)2014-0057,清洁级饲养。经天津中医药大学伦理委员会审查批准通过本次动物实验。
2 方法
2.1 分组、造模及给药
取小鼠48只,随机分为正常组、模型组、甘草附子汤组[7.8 g/kg(以总生药量计算);剂量根据成人临床日常用量,按照实验动物与人体表面积比换算]和雷公藤多苷组(阳性药物组,0.01 g/kg[7]),每组12只。除正常组注射等体积生理盐水外,其余组小鼠均于左后足趾皮内一次性注射弗氏完全佐剂50 μL以建立AA模型[8]。以小鼠造模侧、对侧和前爪足跖明显肿胀,受累关节畸形,行动不便且体质量明显下降判定为造模成功。造模12 d后,剔除造模失败的小鼠后,正常组、模型组小鼠灌胃水,甘草附子汤组和雷公藤多苷组小鼠灌胃相应药液,灌胃体积均为20 mL/kg,每天1次,连续18 d。
2.2 小鼠足肿胀情况观察
分别在造模前和给药结束时,观察并记录小鼠关节病变情況,同时采用水容积法测量小鼠右后足跖容积并计算足肿胀度[肿胀度(mL)=给药结束时足跖容积-造模前足跖容积]。
2.3 小鼠踝关节病理学检查
给药结束后,断颈处死小鼠,于无菌环境下取其右踝关节,石蜡包埋,切片(厚度:5 μm),脱水,HE染色,乙醇梯度脱水,二甲苯透明,中性树胶封片,在显微镜下观察踝关节病理学变化。 [13] 邓伟.甘草附子汤治疗膝骨关节炎的临床研究[J].中药材,2008,31(7):1107-1110.
[14] SUKE SG,NEGI H,MEDIRATTA PK,et al. Anti-arthri- tic and anti-inflammatory activity of combined pioglitazone and prednisolone on adjuvant-induced arthritis[J].Eur J Pharmacol,2013,718(1/2/3):57-62.
[15] ELSHABRAWY HA,CHEN Z,VOLIN MV,et al. The pathogenic role of angiogenesis in rheumatoid arthritis[J].Angiogenesis,2015,18(4):433-448.
[16] YANG X,QIU P,CHEN B,et al. KIAA1199 as a potential diagnostic biomarker of rheumatoid arthritis related to angiogenesis[J]. Arthritis Res Ther,2015. DOI:10.1186/s13075-015-0637-y.
[17] CAO D,HOU M,GUAN YS,et al. Expression of HIF-1α and VEGF in colorectal cancer:association with clinical outcomes and prognostic implications[J]. BMC Cancer,2009. DOI:10.1186/1471-2407-9-432.
[18] ZHANG J,LI C,ZHENG Y,et al. Inhibition of angiogenesis by arsenic trioxide via TSP-1-TGF-β1-CTGF-VEGF functional module in rheumatoid arthritis[J]. Oncotarget,2017,8(43):73529-73546.
[19] 李靜平,顾雯,倪艺榕,等.人参皂苷Rg3对糖尿病大鼠难愈创面表皮细胞及血管新生的影响[J].中国药理学通报,2019,35(4):551-556.
[20] MAIJER KI,NOORT AR,DE HAIR MJ,et al. Nuclear factor-κB-inducing kinase is expressed in synovial endothelial cells in patients with early arthritis and correlates with markers of inflammation:a prospective Cohort study[J]. J Rheumatol,2015,42(9):1573-1581.
[21] NOORT AR,VAN ZOEST KP,WEIJERS EM,et al.NF- kappaB-inducing kinase is a key regulator of inflammation-induced and tumour-associated angiogenesis[J]. J Pathol,2014,234(3):375-385.
[22] HEINEKE J,AUGER-MESSIER M,XU J,et al. Cardiomyocyte GATA4 functions as a stress-responsive regulator of angiogenesis in the murine heart[J]. J Clin Invest,2007,117(11):3198-3210.
[23] JIA W,WU W,YANG D,et al. GATA4 regulates angiogenesis and persistence of inflammation in rheumatoid arthritis[J]. Cell Death Dis,2018,9(5):503-518.
[24] SU CM,HUANG CY,TANG CH. Characteristics of resistin in rheumatoid arthritis angiogenesis[J]. Biomark Med,2016,10(6):651-660.
(收稿日期:2018-12-14 修回日期:2019-05-06)
(编辑:段思怡)