Role of hepatocyte growth factor in rheumatoid arthritis

类风湿关节炎（RA）患者滑液中肝细胞生长因子（HGF）浓度显著升高，且与血清C反应蛋白（CRP）浓度和滑液中IL-6浓度相关，提示HGF在RA的全身性滑膜炎症中起重要作用。在800 x g下离心收集SF细胞，并用磷酸盐缓冲盐水洗涤两次。然后用Ficol-Isopaque梯度分离细胞。将单核细胞部分和多形核细胞部分以5 × 10^6个细胞/ml在补充有10% FCS的RPMI 1640培养基中培养1至3天。将未分级的滑液细胞培养3小时，然后将含有非粘附细胞的培养上清液移至其他培养板。贴壁细胞和非贴壁细胞分别培养1至3天。用酶-I法测定肝细胞生长因子浓度 ...更多信息 墨水免疫吸附测定。仅在多形核细胞部分或贴壁细胞中观察到HGF的时间依赖性分泌。将1，25-二羟维生素D_3（1，25D_3）以10 μ M至10 μ M加入到贴壁细胞培养物中<-11><-6>。1，25 D3以剂量依赖性方式抑制HGF分泌，在10 μ M时观察到最大抑制，产生60%<-7>。另一方面，从27名RA患者、1名MCTD患者、1名SLE患者、1名PM/DM患者、1名Behcet病患者、1名母亲为RA患者的FUO患者和作为正常对照（CTL）的健康志愿者获得骨髓（BM）标本。从胸骨采集1 ml BM标本，用EDTA处理并离心。采用酶联免疫吸附法检测细胞培养上清中HGF和IL-6的含量。RA患者骨髓中HGF水平明显高于CTL。RA患者SF中HGF水平与血清C-反应蛋白呈显著正相关，而与IL-6无显著相关性。提示RA患者SF中HGF是由骨髓中活化的多形核细胞分泌并迁移到SF中的，活性维生素D可能在RA中起免疫调节作用。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