[1]张雪梅,江维.血红素氧合酶-1通过NF-κB信号通路抑制人退变椎间盘髓核细胞凋亡[J].第三军医大学学报,2018,40(07):561-568.
 ZHANG Xuemei,JIANG Wei.Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway[J].J Third Mil Med Univ,2018,40(07):561-568.
点击复制

血红素氧合酶-1通过NF-κB信号通路抑制人退变椎间盘髓核细胞凋亡(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第07期
页码:
561-568
栏目:
基础医学
出版日期:
2018-04-15

文章信息/Info

Title:
Hemeoxygenase-1 protects degenerative human nucleus pulposus cells against apoptosis by NF-κB signaling pathway
作者:
张雪梅江维
重庆医科大学附属第一医院:产科,骨科
Author(s):
ZHANG Xuemei JIANG Wei

Department of Obstetrics, Department of Orthopaedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
血红素氧合酶-1核转录因子kappa BP65髓核凋亡
Keywords:
HO-1NF-&kappaBP65nucleus pulposusapoptosis
分类号:
R681.53; R966; R977.3
文献标志码:
A
摘要:

目的     探讨血红素氧合酶-1(hemeoxygenase-1,HO-1)对人退变椎间盘髓核细胞凋亡的作用及机制。 方法        采用免疫组化和Western blot检测正常(LVF组,标本来源于年轻的椎体骨折行手术治疗的患者)及退变(IDD组,标本来源于椎间盘退变性疾病行手术治疗的患者)人椎间盘髓核组织中HO-1和磷酸化的P65(p-P65)的表达差异;体外培养退变髓核细胞,通过IL-1β处理增加髓核细胞凋亡,再用载HO-1质粒的慢病毒及小干扰RNA处理髓核细胞后检测凋亡的变化;通过P65抑制剂PDTC抑制p-P65表达后检测髓核细胞凋亡的变化。 结果    免疫组化结果显示,HO-1在IDD组中的阳性表达率为(18.58±0.59)%,明显低于LVF组(58.26±2.48)%(P<0.05),而p-P65在IDD组中的阳性表达率为(40.27±2.03)%,明显高于LVF组(19.75±1.98)%(P<0.05)。IL-1β能增加退变髓核细胞的凋亡,同时引起p-P65表达增加,HO-1-siRNA抑制HO-1表达后可以进一步提高髓核细胞凋亡率(P<0.05),而过表达HO-1处理细胞后能够降低IL-1β引起的凋亡增加,同时降低pP65表达(P<0.05)。使用P65抑制剂PDTC抑制髓核细胞P65信号通路后,髓核细胞凋亡率明显降低,此时沉默HO-1不能增加髓核细胞凋亡。 结论      HO-1能够通过NF-κB通路抑制人退变椎间盘髓核细胞凋亡。

Abstract:

Objective    To determine the effect of hemeoxygenase-1 (HO-1) on apoptosis in human degenerative nucleus pulposus cells (NPCs), and investigate the underlying mechanism. Methods     The expression of HO-1 and nuclear factor kappa B (NF-κB) subunit phosphorylated P65 (p-P65) was determined with immunohistochemical assay and Western blot analysis in human nucleus pulposus tissues from healthy young individuals (due to lumbar vertebral fracture, LVF group) and old patients with intervertebral disc degeneration (IDD group). After the degenerative NPCs were cultured in vitro, and IL-1β was added to induce cell apoptosis measured by annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) double staining analysis. The effects of lentiviral vector of HO-1 and its siRNA on the cell apoptosis were also determined. The apoptosis was also observed after the treatment of p65 inhibitor PDTC.  Results     The positive rate of HO-1 was (18.58±0.59)% in the IDD group, significantly lower than that in the LVF group [(58.26±2.48)%, P<0.05], and opposite phenomenon was found in that of p-P65 [(40.27±2.03)% vs (19.75±1.98)%, P<0.05]. IL-1β enhanced the cell apoptosis and up-regulated the expression of p-P65 in the degenerative NPCs. HO-1-siRNA inhibited the HO-1 expression and then promoted cell apoptosis (P<0.05).   Overexpression of HO-1 by LV-HO-1 inhibited the increased NPCs apoptosis after IL-1β treatment, and simultaneously inhibited the expression of p-P65 in human NPCs (P<0.05).  Furthermore, the NF-κB inhibitor, PDTC, was used to treat NPCs,  the number of apoptotic NPCs was significantly decreased no matter with or without HO-1-siRNA transfection. Conclusion      HO-1 inhibits apoptosis in degenerative human NPCs through NF-κB pathway.

参考文献/References:

[1]URBAN J P, ROBERTS S. Degeneration of the intervertebral disc[J].Arthritis Res Ther,2003, 5(3):120-130.
[2]JOUD A, PETERSSON I F, ENGLUND M. Low back pain: epidemiology of consultations[J]. Arthritis Care Res(Hoboken),2012,64(7):1084-1088.DOI: 10.1002/acr.21642.
[3]KIM K W, HA K Y, LEE J S, et al. The apoptotic effects of oxidative stress and antiapoptotic effects of caspase inhibitors on rat notochordal cells[J].Spine,2007,32(22): 2443-2448.DOI: 10.1097/BRS.0b013e318157395a.
[4]POVEDA L, HOTTIGER M, BOOS N, et al. Peroxynitrite induces gene expression in intervertebral disc cells[J]. Spine,2009,34(11): 1127-1133.DOI: 10.1097/BRS.0b013e31819f2330.
[5]ZHAO C Q, WANG L M, JIANG L S, et al.The cell biology of intervertebral discaging and degeneration[J].Ageing Res Rev,2007,6(3):247-261. DOI: 10.1016/j.arr.2007.08.001.
[6]TSUI T Y, LAU C K, MA J, et al.rAAVmediated stable expression of hemeoxygenase-1 in stellate cells: A new approach to attenuate liver fibrosis in rats[J].Hepatology,2005,42(2):335-342. DOI: 10.1002/hep.20803.
[7]LANAVILA A, RODRIGUEZCALVO R, PALOMER X, et al. Atorvastatin inhibits GSK-3β phosphorylation by cardiac hypertrophic stimuli[J]. Biochim Biophys Acta,2008,1781(1/2):26-35. DOI: 10.1016/j.bbalip.2007.10.009.
[8]GUILLER M, MEGISA J, GOMAR F, et al. Haem oxygenase-1 regulates catabolic and anabolic processes in osteoarthritic chondrocytes[J]. J Pathol,2008,214(4):515-522. DOI: 10.1002/path.2313.
[9]HU B, SHI C, XU C, et al. Heme oxygenase-1 attenuates IL-1β induced alteration of anabolic and catabolic activities in intervertebral disc degeneration[J]. Sci Rep, 2016,6:21190. DOI: 10.1038/srep21190.
[10]NAN Y M, WANG R Q, ZHAO S X, et al. Hemeoxygenase-1 prevents non-alcoholic steatohepatitis through suppressing hepatocyte apoptosis in mice[J]. Lipids Health Dis,2010,9(1):124. DOI: 10.1186/1476-511X-9-124.
[11]CAI C, TENG L, VU D, et al. The hemeoxygenase 1 inducer (CoPP) protects human cardiac stem cells against apoptosis through activation of the extracellular signal-regulated kinase (ERK)/NRF2 signaling pathway and cytokine release[J]. J Biol Chem,2012,287(40):33720-33732. DOI: 10.1074/jbc.M112.385542.
[12]WANG X H, HONG X, ZHU L, et al. Tumor necrosis factor alpha promotes the proliferation of human nucleus pulposus cells via nuclear factor-κB, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase[J]. Exp Biol Med,2015,240(4):411-417. DOI: 10.1177/1535370 214554533.
[13]ZHONGYI S, SAI Z, CHAO L,et al. Effects of nuclear factor kappa B signaling pathway in human intervertebral disc degeneration[J]. Spine,2015,40(4): 224-232. DOI: 10.1097/BRS.0000000000000733.
[14]PFIRRMANN C W, METZDORF A, ZANETTI M, et al. Magnetic resonance classification of lumbar intervertebral disc degeneration[J]. Spine,2001,26(17):1873-1878. DOI:10.1097/0000763220010901000011.
[15]WANG D, HU Z, HAO J,et al. SIRT1 inhibits apoptosis of degenerative human disc nucleus pulposus cells through activation of Akt pathway[J]. Age(Dordr),2013,35(5):1741-1753. DOI: 10.1007/s113570129474y.
[16]JIANG W, ZHANG X, HAO J, et al. SIRT1 protects against apoptosis by promoting autophagy in degenerative human disc nucleus pulposuscells[J].Sci Rep,2014,4: 7456. DOI: 10.1038/srep07456.
[17]AHSAN R, TAJIMA N, CHOSA E, et al.Biochemical and morphological changes in herniated human intervertebral disc[J]. J Orthop Sci,2001,6(6):510-518. DOI: 10.1007/s007760100006.
[18]MAINES M D. The hemeoxygenase system and its functions in the brain[J]. Cell Mol Biol,2000,46(3): 573-585.
[19]郭建增,周歧新.脑血红素加氧酶系统的作用研究[J]. 生理科学进展,2002,33(1): 26-29.
GUO J Z, ZHOU Q X. The research into functions of hemeoxygenase system in brain[J]. Prog Physiol Sci,2002,33(1): 26-29.
[20]PONKA P. Tissuespecific regulation of iron metabolism and hemesynthesis: distinct control mechanisms in erythroid cells[J].Blood,1997,89(1): 1-25.
[21]AKI T, YAMAGUCHI K, FUJIMIYA T, et al. Phosphoinositide 3-kinase accelerates autophagic cell death during glucose deprivation in the rat cardiomyocyte-derived cell line H9c2[J].Oncogene,2003,22(52):8529-8535. DOI: 10.1038/sj.onc.1207197
[22]FUKAZAWA R, MILLER T A, KURAMOCHI Y, et al. Neuregulin-1 protects ventricular myocytes from anthracycline-induced apoptosis via erbB4-dependent activation of PI3-kinase/Akt[J]. J Mol Cell Cardiol,2004,35(12):1473-1479.PMID: 14654373. DOI:10.1016/j.yjmcc.2003.09.012.
[23]ZHENG K M, ZHANG J, ZHANG C L, et al. Curcumin inhibits appoptosin-induced apoptosis via up regulating hemeoxygenase-1 expression in SH-SY5Y cells[J]. Acta PharmacolSin,2015, 36(5):544-552. DOI: 10.1038/aps.2014.166.
[24]HAYDEN M S, GHOSH S. Shared principles in NF-kappaB signaling[J]. Cell, 2008,132(2):344-362. DOI: 10.1016/j.cell.2008.01.020.
[25]刘浠,易威威,温亚枫,等.锌指蛋白A20及其相关炎症因子在椎间盘髓核细胞退变前后的表达变化[J].第三军医大学学报,2016,38(18):2077-2081. DOI:10.16016/j.10005404.201602105.
LIU X,YI W W,WEN Y F,et al.Expression of zinc finger protein A20 and related inflammatory factors in nucleus pulposus cells in degenerative intervertebral disc[J].J Third Mil Med Univ,2016,38(18):2077-2081. DOI:10.16016/j.10005404.201602105.
[26]刘刚,唐瑭,吴立兵,等.核因子κB抑制剂Bay 11-7082和131I导致DTC细胞凋亡的效果及协同作用[J].海南医学院学报, 2013,(19)11: 1487-1489. DOI: 10.13210/j.cnki.jhmu.2013.11.036.
LIU G, TANG T, WU L B, et al. Effect of NFκB inhibitor BAY117082 and 131I of inducing apoptosis of cells of differentiated thyroid cancer and their synergistic effects[J]. J Hainan Med Univ, 2013, (19)11: 1487-1489. DOI: 10.13210/j.cnki.jhmu.2013.11.036.
[27]孙中仪,田纪伟.NF-κB信号通路与椎间盘退变的研究进展[J].中国矫形外科杂志,2012,20(23):2162-2164.
SUN Z Y,TIAN J W.Progress in the research of NF-κB signaling pathway and intervertebral disc degeneration[J]. Orthopedic J Chin,20(23):2162-2164.
[28]NASTO L A, SEO H Y, ROBINSON A R,et al . ISSLS prize winner: inhibition of NFκB activity ameliorates ageassociated disc degeneration in a mouse model of accelerated aging[J]. Spine,2012,37(21): 1819. DOI: 10.1097/BRS.0b013e31824ee8f7.
[29]LIU Z, MA C, SHEN J, et al. SDF-1/CXCR4 axis induces apoptosis of human degenerative nucleus pulposus cells via the NF-κB pathway[J]. Mol Med Rep,2016,14(1):783-789. DOI: 10.3892/mmr.2016.5341.

更新日期/Last Update: 2018-04-09