Role of hepatocyte growth factor in rheumatoid arthritis

RA患者滑膜液中肝细胞生长因子（HGF）浓度显著升高，并与血清c反应蛋白浓度和滑膜液中IL-6浓度相关，提示HGF在类风湿性关节炎（RA）全身滑膜炎症中的重要作用。滑液（SF）样本取自RA患者。收集SF细胞，800 x g离心，用磷酸盐缓冲盐水洗涤2次。然后用Ficol-Isopaque梯度分离细胞。单核细胞部分和多形核细胞部分在添加10% FCS的RPMI1640培养基中以5 × 10^6个细胞/ml培养1 ~ 3天。将未分离的滑膜液细胞培养3小时，然后将含有非贴壁细胞的培养上清移至其他培养板。贴壁细胞和非贴壁细胞分别培养1 ~ 3天。采用酶联免疫吸附法测定HGF浓度。仅在多形核细胞部分或贴壁细胞中观察到HGF分泌的时间依赖性。1,25-二羟基维生素D_3 （1,25D_3）在10^<-11> M至10^<-6>M的贴壁细胞培养中加入。1,25 d3对HGF的抑制呈剂量依赖性，在10^<-7>M时最大抑制率为60%。另一方面，取27例RA患者、1例MCTD患者、1例SLE患者、1例PM/DM患者、1例Behcet病患者、1例FUO患者（其母亲为RA患者）和健康志愿者作为正常对照（CTL）的骨髓（BM）标本。取胸骨BM标本1ml， EDTA处理后离心。酶联免疫吸附法测定上清液中HGF和IL-6的浓度。RA患者BM中HGF水平明显高于CTL。RA的HGF水平与血清c反应蛋白浓度显著相关，而与IL-6无关。上述结果提示，RA的SF中HGF与RA活性呈正相关，是由骨髓活化的多形核细胞分泌并迁移到SF中，活性形式的维生素D可能在RA中起免疫调节剂的作用。少

The concentration of Hepatocyte Growth Factor(HGF) significantly increases in synovial fluid obtained from RA patients and it correlated with serum C-reactive protein concentrations and the concentrations of IL-6 in synovial fluid, strongly suggesting the important role of HGF in systemic synovial inflammation in rheumatoid arthritis(RA).Synovial fluid(SF) samples were obtained from patients with RA. SF cells were collected with centrifugation at 800 x g, and washed twice with phosphate buffered saline. Then the cells fractionated with Ficol-Isopaque gradient. Monocyte fraction and polymorphonuclear cell fraction were cultured at 5x10^6 cell/ml in RPMI1640 medium supplemented with 10% FCS for 1 to 3 days. Unfractionated synovial fluid cells were cultured for 3 hours, then culture supernatant containing non-adherent cells were moved to other culture plates. Adherent cells and non adherent cells were cultured separately for 1 to 3 days. The concentrations of HGF were measured by enzyme-l … More inked immunosorbent assay. Time dependent secretion of HGF was observed only in polymorphonuclear cell fraction or adherent cells. 1,25-dihydroxyvitamin D_3 (1,25D_3) was added into the adherent cell culture at 10^<-11> to 10^<-6>M. HGF secretion was suppressed by 1,25D3 in dose dependent manner and the maximum suppression was observed yielding 60% at 10^<-7>M.On the other hand, bone marrow(BM) specimens were obtained from 27 of RA patients, 1 of MCTD, 1 of SLE, 1 of PM/DM, one of Behcet's disease, 1 of FUO, whose mother is an RA patient, and healthy volunteers as normaal controls(CTL). 1 ml of BM specimens sampled from sternal bone and treated with EDTA and centrifuged. The concentrations of HGF and IL-6 in the supernatants were measured by enzyme-linked immunosorbent assay. HGF levels in BM of RA patients were significantly higher than that of CTL. HGF levels of RA were significantly correlated with serum C-reactive protein consentrations but not IL-6.These results suggested that HGF in SF of RA positively correlated with RA activity is secreted by polymorphonuclear cell activated in bone marrow and migrated into SF, and active form of vitamin D may act immmunomodulater in RA. Less