CAJ | 학술논문

目的：探讨瘦素对椎间盘髓核细胞中退行性变相关分解代谢基因的影响，并探讨其机制。方法：培养SD大鼠髓核细胞，行cytokeratin 19和II型胶原免疫组化进行鉴定。使用瘦素和（或）白细胞介素1β（ IL－1β）作用于髓核细胞，real－time PCR分析MMP－1、MMP－3、MMP－9、MMP－13、ADAMTS－4、ADAMTS－5、aggrecan 和COL2A1的表达水平。阿利辛蓝染色和免疫组化分析II型胶原和蛋白多糖的生成。 Western blot 分析激活的信号通路，并使用不同通路的抑制剂来分析信号通路的作用。结果： Real－time PCR显示单用瘦素可以提高MMP－1、MMP－13、ADAMTS－4和ADAMTS－5的表达水平；IL－1β和瘦素可以协同提高MMP－1、MMP－3和ADAMTS－5的表达水平；瘦素降低髓核细胞II型胶原的表达，PI3K／Akt通路和JAK2／STAT3通路均被激活，但使用抑制剂后显示只有JAK2／STAT3信号通路参与瘦素对髓核细胞的作用。结论：瘦素通过调节JAK2／STAT3信号通路促进髓核细胞的分解代谢，可能是肥胖与椎间盘退变相互关联的机制。
목적：탐토수소대추간반수핵세포중퇴행성변상관분해대사기인적영향，병탐토기궤제。방법：배양SD대서수핵세포，행cytokeratin 19화II형효원면역조화진행감정。사용수소화（혹）백세포개소1β（ IL－1β）작용우수핵세포，real－time PCR분석MMP－1、MMP－3、MMP－9、MMP－13、ADAMTS－4、ADAMTS－5、aggrecan 화COL2A1적표체수평。아리신람염색화면역조화분석II형효원화단백다당적생성。 Western blot 분석격활적신호통로，병사용불동통로적억제제래분석신호통로적작용。결과： Real－time PCR현시단용수소가이제고MMP－1、MMP－13、ADAMTS－4화ADAMTS－5적표체수평；IL－1β화수소가이협동제고MMP－1、MMP－3화ADAMTS－5적표체수평；수소강저수핵세포II형효원적표체，PI3K／Akt통로화JAK2／STAT3통로균피격활，단사용억제제후현시지유JAK2／STAT3신호통로삼여수소대수핵세포적작용。결론：수소통과조절JAK2／STAT3신호통로촉진수핵세포적분해대사，가능시비반여추간반퇴변상호관련적궤제。
AIM:To explore the effect of leptin on the expression of degeneration-related genes in rat nucleus pulposus ( NP) cells and to detect the possible mechanism .METHODS:The normal NP cells isolated from SD rats were analyzed by immunochemistry and immunofluorescence for the collagen II and cytokeratin 19 expression.The NP cells were treated with leptin and/or interleukin-1β( IL-β).The mRNA expression of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, ADAMTS-5, aggrecan and COL2A1 in the cells was detected by real-time PCR.Alcian blue staining and im-munochemistry were used to examine the expression of proteoglycan and collagen II .Activation of involved pathways was studied by Western blot .The inhibitors of the pathways were used to reveal the effect of these pathways on NP cells .RE-SULTS:The results of real-time PCR revealed that leptin alone up-regulated the mRNA expression of MMP-1, MMP-13, ADAMTS-4, ADAMTS-5 and COL2A1.The synergy of leptin and IL-βwas found in the increased expression of MMP-1, MMP-3 and ADAMTS-5.The NP cells treated with leptin showed less expression of collagen II .Both PI3K/Akt and JAK2/SATA3 pathways were activated by leptin , whereas only inhibitor of JAK 2/SATA3 pathway reversed the expression of MMP-1 and MMP-13.CONCLUSION:Leptin may promote catabolism in rat NP cells via JAK2/SATA3 pathways, which may be the mechanism mediating the association between obesity and intervertebral disc degeneration .