Promotion of the articular cartilage proteoglycan degradation by T-2 toxin and selenium protective effect  

作　　者：Si-yuan LI Jun-ling CAO Zhong-li SHI Jing-hong CHEN Zeng-tie ZHANG Clare E. HUGHES Bruce CATERSON 

机构地区：[1]Institute of Endemic Diseases, College of Medicine, Xi＇an Jiaotong University Key Laboratory of Environment and Genes Related to Diseases （Xi＇an Jiaotong University）, Ministry of Education Kev Laboratory of Microelement and Endemic Disease （Xi＇an Jiaotong University）, Ministr）, of Health Xi＇an 710061, China [2]Laboratory of Connective Tissue Biology, School of Biosciences, Cardiff University, Cardiff CF10 3US, UK

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2008年第1期22-33,共12页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project supported by the National Natural Science Foundation of China (Nos. 30471499 and 30170831);the Ministry of Education of China (No.Key 03152);the Science Foundation of Shaanxi Province of China (No.2004KW-20)

摘　　要：Objective: To identify the relationship between T-2 toxin and Kashin-Beck disease (KBD),the effects of T-2 toxin on aggrecan metabolism in human chondrocytes and cartilage were investigated in vitro. Methods: Chondrocytes were isolated from human articular cartilage and cultured in vitro. Hyaluronic acid (HA),soluble CD44 (sCD44),IL-1β and TNF-α levels in super-natants were measured by enzyme-linked immunosorbent assay (ELISA). CD44 content in chondrocyte membrane was deter-mined by flow cytometry (FCM). CD44,hyaluronic acid synthetase-2 (HAS-2) and aggrecanases mRNA levels in chondrocytes were determined using reverse transcription polymerase chain reaction (RT-PCR). Immunocytochemical method was used to investigate expressions of BC-13,3-B-3(-) and 2-B-6 epitopes in the cartilage reconstructed in vitro. Results: T-2 toxin inhibited CD44,HAS-2,and aggrecan mRNA expressions,but promoted aggrecanase-2 mRNA expression. Meanwhile,CD44 expression was found to be the lowest in the chondrocytes cultured with T-2 toxin and the highest in control plus selenium group. In addition,ELISA results indicated that there were higher sCD44,IL-1β and TNF-α levels in T-2 toxin group. Similarly,higher HA levels were also observed in T-2 toxin group using radioimmunoprecipitation assay (RIPA). Furthermore,using monoclonal antibodies BC-13,3-B-3 and 2-B-6,strong positive immunostaining was found in the reconstructed cartilage cultured with T-2 toxin,whereas no positive staining or very weak staining was observed in the cartilage cultured without T-2 toxin. Selenium could partly inhibit the effects of T-2 toxin above. Conclusion: T-2 toxin could inhibit aggrecan synthesis,promote aggrecanases and pro-inflammatory cytokines production,and consequently induce aggrecan degradation in chondrocytes. These will perturb metabolism balance between aggrecan synthesis and degradation in cartilage,inducing aggrecan loss in the end,which may be the initiation of the cartilage degradation.Objective： To identify the relationship between T-2 toxin and Kashin-Beck disease （KBD）, the effects of T-2 toxin on aggrecan metabolism in human chondrocytes and cartilage were investigated in vitro. Methods： Chondrocytes were isolated from human articular cartilage and cultured in vitro. Hyaluronic acid （HA）, soluble CD44 （sCD44）, IL-1β and TNF-α levels in supernatants were measured by enzyme-linked immunosorbent assay （ELISA）. CD44 content in chondrocyte membrane was determined by flow cytometry （FCM）. CD44, hyaluronic acid synthetase-2 （HAS-2） and aggrecanases mRNA levels in chondrocytes were determined using reverse transcription polymerase chain reaction （RT-PCR）. Immunocytochemical method was used to investigate expressions of BC-13, 3-B-3（-） and 2-B-6 epitopes in the cartilage reconstructed in vitro. Results： T-2 toxin inhibited CD44, HAS-2, and aggrecan mRNA expressions, but promoted aggrecanase-2 mRNA expression. Meanwhile, CD44 expression was found to be the lowest in the chondrocytes cultured with T-2 toxin and the highest in control plus selenium group. In addition, ELISA results indicated that there were higher sCD44, IL-1β and TNF-α levels in T-2 toxin group. Similarly, higher HA levels were also observed in T-2 toxin group using radioimmunoprecipitation assay （RIPA）. Furthermore, using monoclonal antibodies BC-13, 3-B-3 and 2-B-6, strong positive immunostaining was found in the reconstructed cartilage cultured with T-2 toxin, whereas no positive staining or very weak staining was observed in the cartilage cultured without T-2 toxin. Selenium could partly inhibit the effects of T-2 toxin above. Conclusion： T-2 toxin could inhibit aggrecan synthesis, promote aggrecanases and pro-inflammatory cytokines production, and consequently induce aggrecan degradation in chondrocytes. These will perturb metabolism balance between aggrecan synthesis and degradation in cartilage, inducing aggrecan loss in the end, which may be the initiation of the cartilage degrad

关 键 词：T-2 toxin Kashin-Beck disease （KBD） AGGRECAN IL-1β TNF-α AGGRECANASE Hyaluronic acid （HA） CD44 

分 类 号：R599[医药卫生—内科学]