固醇调节元件结合蛋白2在软骨细胞退变过程中的表达（英文）
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摘要
背景:近期研究证实了固醇调节元件结合蛋白2基因在骨关节炎发生过程中起重要作用,但其具体发病机制尚未完全清楚。目的:通过白细胞介素1β体外诱导关节软骨细胞退变,观察固醇调节元件结合蛋白2在软骨细胞退变过程中的表达变化。方法:体外分离培养C57BL/6J小鼠关节软骨细胞,将第2代软骨细胞随机分为4组:对照组、白细胞介素1β24,48,72 h组。后3组细胞分别以10μg/L白细胞介素1β干预细胞。结果与结论:软骨细胞经白细胞介素1β刺激后呈肥大化表现,软骨细胞活性随白细胞介素1β刺激时间的延长而逐渐降低。与对照组相比,白细胞介素1β24,48,72 h组软骨细胞中固醇调节元件结合蛋白2与固醇调节元件结合蛋白裂解激活蛋白mR NA表达水平增加,而蛋白聚糖和Ⅱ型胶原mR NA表达水平降低。提示白细胞介素1β能抑制软骨细胞增殖和细胞重要基质成分表达,诱导其出现肥大化退行性改变,且在退变的过程中,固醇调节元件结合蛋白2表达逐渐上调,与软骨关键基因的表达呈负向变化关系。
BACKGROUND: Recent studies have demonstrated that sterol regulatory element binding protein-2(SREBP-2) plays a key role in osteoarthritis, but its exact pathogenesis remains incompletely understood yet. OBJECTIVE: To investigate the expression of SREBP-2 in the process of interleukin-1β-induced articular chondrocyte degeneration in vitro. METHODS: Articular chondrocytes obtained from C57BL/6J mice were cultured in vitro. After the second passage, cells were randomly divided into four groups: control group, and three experimental groups treated with 10 μg/L interleukin-1β for 24, 48 and 72 hours, respectively. RESULTS AND CONCLUSION: The cells became hypertrophic after being stimulated by interleukin-1β, and the staining of collagen X was positive at 72 hours. MTT assay demonstrated that the cell activity after stimulation with interleukin-1β decreased with time. Results of RT-PCR showed that the expression of SREBP-2 and SREBP cleavage activating protein m RNA was significantly increased after stimulation with interleukin-1β as compared with the control group and increased with time. On the contrary, the expression of aggrecan and collagen II m RNA was decreased with time. It is revealed that interleukin-1β could inhibit the proliferation of regular chondrocytes and the expression of its extracellular matrix, and furthermore, induce chondrocyte hypertrophy. The expression of SREBP-2 showed a negative relationship with key cartilage genes during this interleukin-1β-induced degeneration.
BACKGROUND: Recent studies have demonstrated that sterol regulatory element binding protein-2(SREBP-2) plays a key role in osteoarthritis, but its exact pathogenesis remains incompletely understood yet. OBJECTIVE: To investigate the expression of SREBP-2 in the process of interleukin-1β-induced articular chondrocyte degeneration in vitro. METHODS: Articular chondrocytes obtained from C57BL/6J mice were cultured in vitro. After the second passage, cells were randomly divided into four groups: control group, and three experimental groups treated with 10 μg/L interleukin-1β for 24, 48 and 72 hours, respectively. RESULTS AND CONCLUSION: The cells became hypertrophic after being stimulated by interleukin-1β, and the staining of collagen X was positive at 72 hours. MTT assay demonstrated that the cell activity after stimulation with interleukin-1β decreased with time. Results of RT-PCR showed that the expression of SREBP-2 and SREBP cleavage activating protein m RNA was significantly increased after stimulation with interleukin-1β as compared with the control group and increased with time. On the contrary, the expression of aggrecan and collagen II m RNA was decreased with time. It is revealed that interleukin-1β could inhibit the proliferation of regular chondrocytes and the expression of its extracellular matrix, and furthermore, induce chondrocyte hypertrophy. The expression of SREBP-2 showed a negative relationship with key cartilage genes during this interleukin-1β-induced degeneration.
引文
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[8]Zeng H,Xiao DM,Tao K,et al.Effects of transforming growth factor-β1 on the expression of Wnt signaling pathway related genes in the differentiation of bone marrow stromal cells.Zhonghua Shiyan Waike Zazhi.2013;30(7):1347-1350.
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[10]Briggs MR,Yokoyama C,Wang X,et al.Nuclear protein that binds sterol regulatory element of low density lipoprotein receptor promoter.I.Identification of the protein and delineation of its target nucleotide sequence.J Biol Chem.1993;268(19):14490-14496.
[11]Rice LM,Donigan M,Yang M,et al.Protein phosphatase 2A(PP2A)regulates low density lipoprotein uptake through regulating sterol response element-binding protein-2(SREBP-2)DNA binding.J Biol Chem.2014;289(24):17268-17279.
[12]Sato R.SREBPs:protein interaction and SREBPs.FEBS J.2009;276(3):622-627.
[13]Salih S,Sutton P.Obesity,knee osteoarthritis and knee arthroplasty:a review.BMC Sports Sci Med Rehabil.2013;5(1):25.
[14]Lee R,Kean WF.Obesity and knee osteoarthritis.Inflammopharmacology.2012;20(2):53-58.
[15]Wang L,Gai P,Xu R,et al.Shikonin protects chondrocytes from interleukin-1beta-induced apoptosis by regulating PI3K/Akt signaling pathway.Int J Clin Exp Pathol.2015;8(1):298-308.
[16]Wang X,Li F,Fan C,et al.Effects and relationship of ERK1and ERK2 in interleukin-1β-induced alterations in MMP3,MMP13,type II collagen and aggrecan expression in human chondrocytes.Int J Mol Med.2011;27(4):583-589.
[17]Smith SM,Shu C,Melrose J.Comparative immunolocalisation of perlecan with collagen II and aggrecan in human foetal,newborn and adult ovine joint tissues demonstrates perlecan as an early developmental chondrogenic marker.Histochem Cell Biol.2010;134(3):251-263.
[18]Li LC,Varghese Z,Moorhead JF,et al.Cross-talk between TLR4-MyD 88-NF-κB and SCAP-SREBP2 pathways mediates macrophage foam cell formation.Am J Physiol Heart Circ Physiol.2013;304(6):H874-884.
[19]Hamamura K,Lin CC,Yokota H.Salubrinal reduces expression and activity of MMP13 in chondrocytes.Osteoarthritis Cartilage.2013;21(5):764-772.
[2]Sorour AS,Ayoub AS,Abd El Aziz EM.Effectiveness of acupressure versus isometric exercise on pain,stiffness,and physical function in knee osteoarthritis female patients.J Adv Res.2014;5(2):193-200.
[3]Kerkhoffs GM,Servien E,Dunn W,et al.The influence of obesity on the complication rate and outcome of total knee arthroplasty:a meta-analysis and systematic literature review.J Bone Joint Surg Am.2012;94(20):1839-1844.
[4]Shao W,Espenshade PJ.Sterol regulatory element-binding protein(SREBP)cleavage regulates Golgi-to-endoplasmic reticulum recycling of SREBP cleavage-activating protein(SCAP).J Biol Chem.2014;289(11):7547-7557.
[5]Liu FH,Song JY,Shang XR,et al.The gene-gene interaction of INSIG-SCAP-SREBP pathway on the risk of obesity in Chinese children.Biomed Res Int.2014;2014:538564.
[6]Kostopoulou F,Gkretsi V,Malizos KN,et al.Central role of SREBP-2 in the pathogenesis of osteoarthritis.PLo S One.2012;7(5):e35753.
[7]Buckwalter JA,Anderson DD,Brown TD,et al.The roles of mechanical stresses in the pathogenesis of osteoarthritis:implications for treatment of joint injuries.Cartilage.2013;4(4):286-294.
[8]Zeng H,Xiao DM,Tao K,et al.Effects of transforming growth factor-β1 on the expression of Wnt signaling pathway related genes in the differentiation of bone marrow stromal cells.Zhonghua Shiyan Waike Zazhi.2013;30(7):1347-1350.
[9]Qiu XM,Jin CT,Wang W.Association between single nucleotide polymorphisms of sterol regulatory element binding protein-2 gene and risk of knee osteoarthritis in a Chinese Han population.J Int Med Res.2014;42(2):320-328.
[10]Briggs MR,Yokoyama C,Wang X,et al.Nuclear protein that binds sterol regulatory element of low density lipoprotein receptor promoter.I.Identification of the protein and delineation of its target nucleotide sequence.J Biol Chem.1993;268(19):14490-14496.
[11]Rice LM,Donigan M,Yang M,et al.Protein phosphatase 2A(PP2A)regulates low density lipoprotein uptake through regulating sterol response element-binding protein-2(SREBP-2)DNA binding.J Biol Chem.2014;289(24):17268-17279.
[12]Sato R.SREBPs:protein interaction and SREBPs.FEBS J.2009;276(3):622-627.
[13]Salih S,Sutton P.Obesity,knee osteoarthritis and knee arthroplasty:a review.BMC Sports Sci Med Rehabil.2013;5(1):25.
[14]Lee R,Kean WF.Obesity and knee osteoarthritis.Inflammopharmacology.2012;20(2):53-58.
[15]Wang L,Gai P,Xu R,et al.Shikonin protects chondrocytes from interleukin-1beta-induced apoptosis by regulating PI3K/Akt signaling pathway.Int J Clin Exp Pathol.2015;8(1):298-308.
[16]Wang X,Li F,Fan C,et al.Effects and relationship of ERK1and ERK2 in interleukin-1β-induced alterations in MMP3,MMP13,type II collagen and aggrecan expression in human chondrocytes.Int J Mol Med.2011;27(4):583-589.
[17]Smith SM,Shu C,Melrose J.Comparative immunolocalisation of perlecan with collagen II and aggrecan in human foetal,newborn and adult ovine joint tissues demonstrates perlecan as an early developmental chondrogenic marker.Histochem Cell Biol.2010;134(3):251-263.
[18]Li LC,Varghese Z,Moorhead JF,et al.Cross-talk between TLR4-MyD 88-NF-κB and SCAP-SREBP2 pathways mediates macrophage foam cell formation.Am J Physiol Heart Circ Physiol.2013;304(6):H874-884.
[19]Hamamura K,Lin CC,Yokota H.Salubrinal reduces expression and activity of MMP13 in chondrocytes.Osteoarthritis Cartilage.2013;21(5):764-772.