Tulamo R M
College of Veterinary Medicine, Department of Clinical Sciences, Helsinki, Finland.
Am J Vet Res. 1991 Dec;52(12):1940-4.
Two recently developed direct methods, radioassay-125I-labeled hyaluronic acid binding protein (125I-HABP)- and high-performance liquid chromatography (HPLC), were used to assess and compare the concentration of hyaluronate (HA) in synovial fluid of horses. Also determined were changes in the HA concentration in an experimental treatment model involving physiologic saline solution (PSS)-irrigated or methylprednisolone acetate-injected tarsocrural joints of clinically normal horses. Serum HA concentration was determined simultaneously, using the 125I-HABP assay. Synovial fluid HA concentration values obtained by use of the HPLC method were approximately double the values obtained by use of 125I-HABP assay. Correlation (r = 0.819) between the 2 methods was highly significant (P less than 0.001; linear regression analysis) for all samples studied and for various experimental subgroups. When pure HA standards were used, correlation between the 2 methods was close to 1 (r = 0.965; P less than 0.001), with higher values obtained by use of the 125I-HABP assay. It is suggested that the HA binding protein derived from endogenous cartilage proteoglycan interferes with the 125I-HABP assay on synovial fluid, resulting in excessively low values, compared with those obtained using the HPLC procedure. Intra-articular injection of methylprednisolone acetate significantly (P less than 0.01) increased synovial fluid HA concentration at 24 hours after injection. Increase was also detected after PSS irrigation, but owing to wide intersubject variation, this increase was not significant. The HPLC procedure, which provides simultaneous information about the concentration and degree of polymerization of HA, is recommended for the study of synovial fluid, whereas the 125I-HABP assay is more suitable for serum HA analysis.
两种最近开发的直接方法，即放射分析法——125I标记的透明质酸结合蛋白（125I-HABP）法和高效液相色谱法（HPLC），被用于评估和比较马滑液中透明质酸盐（HA）的浓度。同时还测定了在一个实验性治疗模型中，临床正常马的跗关节经生理盐水（PSS）冲洗或注射醋酸甲泼尼龙后HA浓度的变化。使用125I-HABP分析法同时测定血清HA浓度。通过HPLC法获得的滑液HA浓度值约为使用125I-HABP分析法所获值的两倍。对于所有研究样本和各个实验亚组，两种方法之间的相关性（r = 0.819）非常显著（P < 0.001；线性回归分析）。当使用纯HA标准品时，两种方法之间的相关性接近1（r = 0.965；P < 0.001），125I-HABP分析法所获值更高。提示内源性软骨蛋白聚糖衍生的HA结合蛋白干扰了滑液的125I-HABP分析，导致与使用HPLC法所获值相比过低的值。关节内注射醋酸甲泼尼龙在注射后24小时显著（P < 0.01）增加了滑液HA浓度。PSS冲洗后也检测到增加，但由于个体间差异较大，这种增加不显著。HPLC法能同时提供HA浓度和聚合度的信息，推荐用于滑液研究，而125I-HABP分析法更适合血清HA分析。
