Du Y, Wang J W, Zhou M J, Wu X Y, Xue J, Wu H X
Department of Rheumatology, Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China.
Zhonghua Nei Ke Za Zhi. 2018 Sep 1;57(9):661-666. doi: 10.3760/cma.j.issn.0578-1426.2018.09.008.
To investigate the clinical significance of soluble Semaphorin 5A(Sema 5A) in patients of rheumatoid arthritis(RA) and the effect of Sema 5A on osteoclastogenesis in RAW264.7 cells. (1)Soluble Sema 5A was detected in the serum of 62 RA patients, 30 osteoarthritis(OA) patients and 48 healthy controls(HC) by enzyme-linked immunosorbent assay(ELISA).The relationships between serum Sema 5A and disease activity, radiographic severity and laboratory parameters were investigated in RA patients.(2)RAW264.7 cells were treated with different concentrations of Sema 5A(0,0.5,1,2.5,5 μg/ml).After 7 days, tartrate-resistant acid phosphate(TRAP) staining was performed. The mRNA levels of TRAP, cathepsin-K and matrix metallopeptidase 9(MMP-9) were tested using real-time polymerase chain reaction (RT-PCR).(3)Bone resorption area of dentine slides cultured with RAW264.7 cells was calculated after Sema 5A(5μg/ml) treatment. (1)The serum Sema 5A in RA patients[(5.24±0.59)μg/L] was significantly higher than those in healthy controls[(2.93±0.34)μg/L,0.01] and OA patients[(2.68±0.47)μg/L,0.05]. The Sema 5A level in RA patients was positively correlated with disease activity score with 28 joint using C-reactive protein(DAS28-CRP), clinical disease activity index (CDAI),C-reactive protein(CRP) and sharp scores(<0.05 or 0.01).In addition,the serum Sema 5A in RA patients with positive anti-cyclic citrullinated peptide(CCP) antibody was significantly greater than that of anti-CCP antibody-negative patients(0.05).(2)After RAW264.7 cells were treated with Sema 5A, the number of TRAP positive osteoclasts increased according to the increase of Sema 5A concentration with maximal effect at 5 μg/ml. Meanwhile, Sema 5A promoted mRNA expression of TRAP,Cathepsin-K and MMP-9.(3)Bone resorption area increased when RAW264.7 cells were treated with Sema 5A(5 μg/ml). Serum Sema 5A is elevated in RA patients and correlated with disease activity and radiographic severity. Sema 5A promotes osteoclastogenesis of RAW264.7 cells.
探讨可溶性信号素5A（Sema 5A）在类风湿关节炎（RA）患者中的临床意义以及Sema 5A对RAW264.7细胞破骨细胞生成的影响。（1）采用酶联免疫吸附测定（ELISA）法检测62例RA患者、30例骨关节炎（OA）患者及48例健康对照者（HC）血清中的可溶性Sema 5A。研究RA患者血清Sema 5A与疾病活动度、影像学严重程度及实验室指标之间的关系。（2）用不同浓度的Sema 5A（0、0.5、1、2.5、5μg/ml）处理RAW264.7细胞。7天后，进行抗酒石酸酸性磷酸酶（TRAP）染色。采用实时聚合酶链反应（RT-PCR）检测TRAP、组织蛋白酶K和基质金属蛋白酶9（MMP-9）的mRNA水平。（3）计算Sema 5A（5μg/ml）处理后RAW264.7细胞培养的牙本质玻片的骨吸收面积。（1）RA患者血清Sema 5A[（5.24±0.59）μg/L]显著高于健康对照者[（2.93±0.34）μg/L，P<0.01]和OA患者[（2.68±0.47）μg/L，P<0.05]。RA患者Sema 5A水平与采用C反应蛋白的28个关节疾病活动评分（DAS28-CRP）、临床疾病活动指数（CDAI）、C反应蛋白（CRP）及Sharp评分呈正相关（P<0.05或P<0.01）。此外，抗环瓜氨酸肽（CCP）抗体阳性的RA患者血清Sema 5A显著高于抗CCP抗体阴性患者（P<0.05）。（2）RAW264.7细胞经Sema 5A处理后，TRAP阳性破骨细胞数量随Sema 5A浓度增加而增多，5μg/ml时作用最强。同时，Sema 5A促进TRAP、组织蛋白酶K和MMP-9的mRNA表达。（3）RAW264.7细胞经Sema 5A（5μg/ml）处理后骨吸收面积增加。RA患者血清Sema 5A升高，且与疾病活动度和影像学严重程度相关。Sema 5A促进RAW264.7细胞的破骨细胞生成。
