ABSTRACT
Objective To identify differently expressed genes and pathways between Kashin-Beck disease (KBD) cartilage and healthy cartilage,and to explore the mechanism of articular cartilage lesions of KBD.Methods Cartilage specimens were collected from 9 patients with KBD and 9 healthy controls.Total RNA was extracted from cartilage specimens,and transcribed into cDNA.KBD and control groups were labeled by Cy3 and Cy5,respectively.Agilent genome-wide microarray was applied to compare the expression profile of KBD cartilage and healthy cartilage.The microarray data was analyzed by single gene and pathway expression analysis to identify differently expressed genes and pathways between KBD and healthy controls.Results ①Tweenty nine genes were significantly up-regulated in KBD group (averaged ratio =6.68 + 1.98,P ＜ 0.05),mainly involved in apoptosis,metabolism,extracellular matrix,cytoskeleton and cell movement.Additionally,extracellular matrix-related FBLN1 gene was down-regulated in KBD group(ratio =0.14 + 0.06,P ＜ 0.05).②Five apoptosis and 6 hypoxia-related pathways presented higher expression levels in KBD compared to healthy controls(all P＜ 0.05).Conclusions We find significant expression differences of apoptosis and hypoxia-related genes and pathways between KBD cartilages and healthy cartilages,suggesting that hypoxia might contribute to chondrocytes apoptosis of KBD.Further studies may be needed to investigate the relationship between hypoxia and articular cartilage lesions of KBD.
ABSTRACT
Objective To compare the expression profile of mycotoxin-related environmental response genes (MERGs) in the articular cartilage of patients with Kashin-Beck disease (KBD) and healthy controls,and explore the relationship between MERG and KBD.Methods Articular cartilage specimens were collected from 9 healthy human subjects and 9 adult KBD patients.Agilent microarray was used to evaluate the expression levels of MERG in cartilage specimens,and the expression ratios of MERG between KBD and healthy controls were calculated.GSEA software was used to calculate the NES scores and P values of gene ontology(GO).Results ①T-2 toxin,deoxynivalenol,zearalenone,aflatoxin B1,fumonisin B1 and ochratoxin A related 15 MERGs presented expression differences between KBD and healthy controls(ratios ＞ 2.0 or ＜ 0.5).Thirteen MERGs were up-regulated in KBD,including BAX,BCL2,COL5A2,FER1L3,GSTT2,IGFBP2,IGFBP4,PDE8B,SOCS3,THBS1,TMSL8,VGLL3 and TUBB2A (ratio ＞ 2.0).Two MERGs,POSTN and FABP4,were down-regulated in KBD (ratio ＜ 0.5).The 15 MERGs were involved in various biological processes; such as collage synthesis,apoptosis,metabolism,growth & development and so on.②Mycotoxin related 4 apoptosis GOs and 5 growth & development related GOs were up-regulated in KBD compared to healthy controls(NES ＞ 0),including ANTI_APOPTOSIS,REGULATION_OF_PROGRAMMED_CELL_DEATH,APOPTOSIS_GO,REGULATION_OF_APOPTOSIS,ORGAN_MORPHOGENESIS,ANATOMICAL_STRUCTURE_DEVELOPMENT,ORGAN_DEVELOPMENT,SYSTEM_DEVELOPMENT and REGULATION OF DEVELOPMENTAL_PROCESS (NES ＞ 0 and P ＜ 0.05).Conclusions There are multiple mycotoxins related environmental response genes presenting significant expression difference between KBD cartilage and normal cartilage.Mycotoxin can affect the expression of MERGs in KBD articular cartilage,which might lead to dysfunction of chondrocytes,and articular cartilage lesions.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes in the expressions of Fas-associated death domain protein (FADD) and cellular-FLICE inhibitory protein (c-FLIP) in the articular cartilage of patients with Kashin-Beck disease (KBD) and the role of these proteins in the pathogenesis of KBD.</p><p><b>METHODS</b>The cartilage samples were collected from patients with established diagnosis of KBD and osteoarthritis and from healthy control subjects undergoing amputation due to traffic accidents. The expressions of Fas-associated death domain protein (FADD) and cellular-FLICE inhibitory protein (c-FLIP) in the cartilage were detected by immunohistochemistry, and the positive chondrocytes were counted in different layers of the articular cartilage under microscope.</p><p><b>RESULTS</b>The positivity rates of FADD in the middle layer of articular cartilage from patients with KBD [(28.68∓2.19)%] and osteoarthritis [(35.40∓2.34)%] were significantly higher than that in normal cartilage [(10.51∓5.02)%, F=16.245, P=0.000], but the rates in the upper and deeper layers were comparable among the 3 groups (P=0.206-0.761). In KBD cartilage, FADD expression was the highest in the middle layer [(28.68∓5.38)%] followed by the deeper layer [(17.94∓8.38)%]. Compared with the healthy controls, KBD and osteoarthritis patients showed significantly higher FLIP expression in the upper layer of the cartilage (F=5.929, P=0.018) but similar expressions in middle and deeper layers.</p><p><b>CONCLUSIONS</b>KBD patients have significant increased FADD expression in the middle layer but decreased FLIP expression in the upper layer of the cartilage, suggesting that the death receptor pathway and its regulators play important roles in the pathogenesis of KBD.</p>