成纤维细胞生长因子4对类风湿关节炎成纤维样滑膜细胞增殖的影响  

Effects of fibroblast growth factor 4 on the proliferation of fibroblast-like synoviocytes in rheumatoid arthritis

作　　者：冯晓雪[1] 朱尚玲[2] 刘芳 陈应镝 何石萍 黄建林[2] Feng Xiaoxue;Zhu Shangling;Liu Fang;Chen Yingdi;He Shiping;Huang Jianlin(Department of rheumatology,the Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院风湿免疫科,广州510630 [2]中山大学附属第六医院风湿免疫科

出　　处：《中华风湿病学杂志》2018年第11期768-773,共6页Chinese Journal of Rheumatology

基　　金：国家自然科学基金(81571584);广东省社会发展领域科技计划项目(2013B021800076).

摘　　要：目的观察RA患者血清中成纤维细胞生长因子4(FGF4)的表达情况,并初步探讨FGF4对RA成纤维样滑膜细胞(FLS)增殖的影响及可能机制。方法①收集20例病情活动RA患者及20名与其年龄和性别相匹配的健康对照者的血清标本,蛋白芯片法检测血清FGF4的表达水平。②收集5例病情活动RA患者的滑膜组织,组织块法培养RA-FLS。采用重组人FGF4蛋白(rhFGF4)处理RA-FLS,应用CCK-8法检测细胞增殖率,流式细胞术检测细胞周期分布,蛋白质印迹法检测cyclinD1、phospho-Akt(p-Akt)及phospho-p38(p-p38)蛋白表达水平。2组间比较采用t检验,多组间比较采用单因素方差分析或Kruskal-Wallis检验,两两比较采用Dunnett′s t检验,以P<0.05为差异有统计学意义。结果①活动性RA患者血清中FGF4表达水平高于健康对照组(P=0.041)。②经不同浓度rhFGF4(分别为12.5、25、50、100ng/ml)处理后,RA-FLS的细胞增殖率分别为:(121±8)%、(126±12)%、(129±12)%、(134±14)%,均高于对照组(100±0)%(P12.5=0.049,P25=0.009,P50=0.004,P100=0.001),G2/M+S期细胞所占的比例分别为:(12.6±3.6)%、(15.3±4.5)%、(17.1±5.1)%、(19.6±4.1)%,除12.5ng/ml组外,其余各组G2/M+S期细胞所占的比例均高于对照组(5.4±2.4)%(P12.5=0.159,P25=0.042,P50=0.018,P100=0.005)。与对照组相比,50ng/ml组、100ng/ml组的cyclinD1蛋白表达上调(P50=0.035,P100=0.027)。浓度为50ng/ml的rhFGF4可短时间内引起p-Akt和p-p38蛋白表达增加(P<0.01)。结论RA患者血清中FGF4表达增高,FGF4可能通过PI3K/Akt通路及p38-MAPK通路参与了RA-FLS增殖的调控,从而促进滑膜增生。Objective To investigate the expression of fibroblast growth factor 4 (FGF4) in serum of active rheumatoid arthritis (RA) and its role in RA synoviocyte proliferation. Methods The serum level of FGF4 were detected by protein arrays in 20 patients with RA, and 20 age and gender matched healthy controls. FLSs were isolated from RA synovium, and were co-cultured with recombinant human FGF4 (rhFGF4). Cell proliferation was quantified by Cell Counting Kit-8 assay and cell cycle distribution was evaluated by flow-cytometry. The protein levels of cyclin D1, phospho-Akt (p-Akt) and phospho-p38 (p-p38) were measured by western blot. Results The serum expression of FGF4 in RA group was higher than that in control group (P=0.041). After being treated with different concentrations of rhFGF4 (12.5, 25, 50, and 100 ng/ml), RA-FLS showed significant increase in cell proliferation, with different rates of [(121±8)%], [(126±12)%], [(129±12)%], and [(134±14)%] respectively, comparing with that of the controls [(100±0)%, (P12.5=0.049, P25=0.009, P50=0.004, P100=0.001).]. Among them, the percentage of G2/M+S phase cells were [(12.6±3.6)%], [(15.3±4.5)%], [(17.1±5.1)%], [(19.6±4.1)%] respectively, and except the lowest rhFGF4 concentration treatment group of 12.5 ng/ml, G2/M+S phase cells in other groups was significantly increased compared with the controls [(5.4±2.4)%] (P12.5=0.159, P25=0.042, P50=0.018, P100=0.005). And the protein expression of cyclin D1 was up-regulated after being treated with 50 ng/ml and 100 ng/ml rhFGF4 (P50=0.035, P100=0.027). FGF4 transiently increased the expression of p-Akt and p-p38 protein at the concentration of 50 ng/ml. Comparisons of data between groups were performed by independent sample Student's t-test. Statistical significant differences among groups were tested by one-way analysis of variance (ANOVA) or the Kruskal-Wallis test. The Dunnett's t-test was used for multiple comparisons. A P-value of <0.05 was considered statistically significant. Conclusion Our results suggest that FGF4

关 键 词：关节炎 类风湿 成纤维细胞生长因子4 成纤维样滑膜细胞 细胞增殖 

分 类 号：R593.22[医药卫生—内科学]