杨召;苑珍珍;张杨;郭悦;马剑雄;马信龙;

• 1:天津医科大学总医院骨科
• 2:天津医院骨科

摘要(Abstract)：

[目的]观察不同pH值对体外培养人退变椎间盘髓核细胞凋亡的影响,为椎间盘退变的预防与治疗提供理论依据。[方法]对体外培养的人退变椎间盘髓核细胞利用甲苯胺蓝和番红O染色进行鉴定,并对P3代细胞分别用普通培养基(pH值7.4)、酸性培养基(pH值6.8)和酸性培养基(pH值6.8)+Cucl_2(酸质子受体抑制剂)培养24 h,用流式细胞仪测定细胞凋亡,计算细胞凋亡率。[结果]细胞染色结果均为阳性,证明实验中所培养的细胞为髓核类软骨细胞。普通培养基组的凋亡率为(8.86±0.20)%,酸性培养基组的凋亡率为(11.23±0.77)%,酸性培养基+Cucl_2组的凋亡率为(9.60±0.49)%。与其他两组相比,酸性培养基组的凋亡率明显增加(P<0.05)。[结论]在低pH值的环境下,人退变椎间盘髓核细胞的凋亡率增加。

关键词(KeyWords)： pH值;人髓核细胞;凋亡;椎间盘;退变

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(编号:81572154);; 天津市科技支撑重点项目(编号:13ZCZDSY01700);; 天津市卫生局科技攻关项目(编号:15KG123)

作者(Author): 杨召;苑珍珍;张杨;郭悦;马剑雄;马信龙;

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参考文献(References)：

• [1]Yang W,Yu XH,Wang C,et al.Interleukin-1βin intervertebral disk degeneration[J].Clin Chim Acta,2015,450:262-272.
• [2]Ye S,Ju B,Wang H,et al.Bone morphogenetic protein-2 provokes interleukin-18-induced human intervertebral disc degeneration[J].Bone Joint Res,2016,5(9):412-418.
• [3]Wang F,Cai F,Shi R,et al.Aging and age related stresses:a senescence mechanism of intervertebral disc degeneration[J].Osteoarthritis Cartilage,2016,24(3):398-408.
• [4]Samartzis D,Borthakur A,Belfer I,et al.Novel diagnostic and prognostic methods for disc degeneration and low back pain[J].Spine J,2015,15(9):1919-1932.
• [5]Gilbert HT,Hodson N,Baird P,et al.Acidic p H promotes intervertebral disc degeneration:acid-sensing ion channel-3 as a potential therapeutic target[J].Sci Rep,2016,6:37360.
• [6]Ohshima H,Urban JP.The effect of lactate and p H on proteoglycan and protein synthesis rates in the intervertebral disc[J].Spine(Phila Pa 1976),1992,17(9):1079-1082.
• [7]Horner HA,Urban JP.Volvo Award Winner in Basic Science Studies:effect of nutrient supply on the viability of cells from the nucleus pulposus of the intervertebral disc[J].Spine(Phila Pa),1976,2001,26(23):2543-2549.
• [8]Fernandez C,Marionneaux A,Gill S,et al.Biomimetic nucleus pulposus scaffold created from bovine caudal intervertebral disc tissue utilizing an optimal decellularization procedure[J].J Biomed Mater Res A,2016,104(12):3093-3106.
• [9]Louman-Gardiner KM,Coombe D,Hunter CJ.Computation models simulating notochordal cell extinction during early ageing of an intervertebral disc[J].Comput Methods Biomech Biomed Engin,2011,14(12):1071-1077.
• [10]Malandrino A,Noailly J,Lacroix D.The effect of sustained compression on oxygenn metabolic transport in the intervertebral discdecreases with degenerative changes[J].PLo S Comput Biol,2011,7(8):e1002112.
• [11]Grunhagen T,Wilde G,Soukane DM,et a1.Nutrient supply and intervertebral disc metabolism[J].J Bone Joint Surg Am,2006,88(Suppl 2):30-35.
• [12]Razaq S,Wilkins RJ,Urban JP.The effect of extracellular p H on matrix turnover by cells of the bovine nucleus pulposus[J].Eur Spine J,2003,12(4):341-349.
• [13]Diamant B,Karlsson J,Nachemson A.Correlation between lactate levels and p H in discs of patients with lumbar rhizopathies[J].Experientia,1968,24(12):1195-1196.
• [14]Kitano T,Zerwekh JE,Usui Y,et a1.Biochemical changes associated witht the symptomatic human intervertebral disk[J].Clin Orthop Relat Res,1993,293(293):372-377.
• [15]Crowley LC,Marfell BJ,Scott AP,et a1.Quantitation of apoptosis and necrosis by annexin v binding,propidium iodide uptake,and flow cytometry[J].Cold Spring Harb Protoc,2016,2016(11):pdb.prot087288.
• [16]Nancy SK,Yao ZQ,Kelly KS,et al.Increased bone density in mice lacking the proton receptor,OGR1[J].Kidney Int,2016,89(3):565-573.