Popko Janusz, Marciniak Justyna, Ilendo Elzbieta, Knas Malgorzata, Guszczyn Tomasz, Stasiak-Barmuta Anna, Moniuszko Tadeusz, Zwierz Krzysztof, Wysocka Jolanta
Department of Pediatric Orthopedics and Traumatology, Medical University, Bialystok, Poland.
Cell Biochem Biophys. 2008;51(2-3):89-95. doi: 10.1007/s12013-008-9018-3. Epub 2008 Jul 1.
Determining the activity of lysosomal exoglycosidases in tissue cultures of synoviocytes derived from the knee joints of patients with injured anterior cruciate ligaments (ACL), juvenile idiopathic arthritis (JIA), and rheumatoid arthritis (RA).
The following exoglycosidases in cultured synoviocytes were analyzed with p-nitrophenyl derivatives of appropriate sugars as substrates: hexosaminidase (HEX) and its isoenzyme A (HEX-A), beta-glucuronidase (GluA), beta-galactosidase (GAL), alpha-mannosidase (MAN), and alpha-fucosidase (FUC).
In our cell cultures, fibroblast-like synovial cells (FLS) dominated. In the group of patients with ACL-injuries, and in the groups of patients with JIA and RA, the activity of the investigated exoglycosidases was significantly higher in the intra- rather than in the extracellular compartment. Hexosaminidase was the predominant exoglycosidase. Stimulation of synoviocytes by IL-1beta in cell cultures significantly increased the activity of HEX, HEX-A, and GluA in both compartments, as well as of GAL, MAN, and FUC in the intracellular compartment. Stimulation by IL-1beta rheumatoidal synoviocytes increased by 128-201% the activity of HEX and HEX A in intracellular compartments and 33-72% in extracellular compartment.
The profile of lysosomal exoglycosidases in a cell culture of human synoviocytes is similar, but not identical, to those in the knee joint. Hexosaminidase is the dominant glycosidase in cultured unstimulated and IL-1beta-stimulated human synoviocytes. The HEX inhibitors may be new drugs for the treatment of inflamed knee joints.
测定前交叉韧带(ACL)损伤患者、幼年特发性关节炎(JIA)患者和类风湿关节炎(RA)患者膝关节滑膜细胞组织培养中溶酶体外切糖苷酶的活性。
以适当糖类的对硝基苯基衍生物为底物,分析培养的滑膜细胞中的以下外切糖苷酶:己糖胺酶(HEX)及其同工酶A(HEX-A)、β-葡萄糖醛酸酶(GluA)、β-半乳糖苷酶(GAL)、α-甘露糖苷酶(MAN)和α-岩藻糖苷酶(FUC)。
在我们的细胞培养物中,成纤维样滑膜细胞(FLS)占主导。在ACL损伤患者组以及JIA和RA患者组中,所研究的外切糖苷酶在细胞内而非细胞外区室中的活性显著更高。己糖胺酶是主要的外切糖苷酶。细胞培养中IL-1β刺激滑膜细胞显著增加了两个区室中HEX、HEX-A和GluA的活性,以及细胞内区室中GAL、MAN和FUC的活性。IL-1β刺激类风湿滑膜细胞使细胞内区室中HEX和HEX A的活性增加了128 - 201%,细胞外区室中增加了33 - 72%。
人滑膜细胞培养物中溶酶体外切糖苷酶的谱与膝关节中的相似,但不完全相同。己糖胺酶是培养的未刺激和IL-1β刺激的人滑膜细胞中的主要糖苷酶。HEX抑制剂可能是治疗膝关节炎症的新药。