张 宇,唐小蓓,穆小平,余城墙,吴有财,韦建勋.青少年特发性脊柱侧凸融合术后远端附加现象的研究进展[J].中国脊柱脊髓杂志,2019,(7):656-660. |
青少年特发性脊柱侧凸融合术后远端附加现象的研究进展 |
中文关键词: 髓核细胞 酸环境 卵巢癌G蛋白偶联受体1 自噬 细胞功能 大鼠 |
中文摘要: |
随着矫形技术的发展以及临床医生对特发性脊柱侧凸(adolescent idiopathic scoliosis,AIS)研究的逐渐深入,AIS通过手术矫形的效果得到了显著提高。但脊柱外科医师在对AIS患者术后随访时发现,部分患者在术后融合节段下方出现了弯曲加重的表现,这就是脊柱侧凸矫形术后远端附加现象。
远端附加现象是AIS患者行选择性胸弯融合术治疗后常见的一种冠状面失平衡现象,多见于行选择性胸弯融合的患者,主要是Lenke 1A型以及Lenke 2A型AIS患者。文献报道其术后发生率为12.9%~51.1%。且远端附加现象的定义因研究的不同而异,并应用不同的参数和标准来确定远端附加现象。Sponseller等认为,相对于术后即刻、随访过程中,在原发弯的远端或近端出现侧凸跨度延长,椎体叠加且角度增加6°以上为远端附加现象。而Schlechter等定义远端附加现象为在融合节段以下Cobb角增加,或Cobb角内包含的椎体数量的增加,或者融合节段远端的椎间盘弯曲角度的增加。2011年Wang等将远端附加现象定义为结构性主弯融合术后随访1年及以上时,主弯范围延长,最下固定椎(lowest instrumented vertebra,LIV)下方有更多椎体进入主弯,LIV以下第一个椎体偏离骶骨中垂线(center sacral vertical line,CSVL)5mm以上或LIV以下第一个椎间隙成角增加5°以上的现象。Wang等认为其诊断标准为末次随访时,对比术后站立位脊柱全长正位片出现:(1)主弯下端椎体(lower end vertebra,LEV)向远端移动,主弯椎体数量增加,LIV偏离CSVL>10mm;(2)LIV以下第一个椎体偏离CSVL>5mm;(3)LIV以下第1个椎间隙成角增加>5°。为使更多的脊柱外科医生对该病有充分的认识,避免AIS患者术后远端附加现象的发生,笔者从远端附加现象的发生机制、危险因素、防治等方面对其进行综述。 |
Update on adding-on phenomenon after surgical treatment of adolescent idiopathic scoliosis |
英文关键词:Nucleus pulposus cells Acidic environment Ovarian cancer G-protein-coupled receptor 1 Auophagy Cellular function Rat |
英文摘要: |
【Abstract】 Objectives: To explore the autophagy activation and cell function of nucleus pulposus cells(NPCs) effected by acid environment, through observing the expression levels of ovarian cancer G-protein-coupled re?鄄ceptor 1(OGR1) and autophagy-related protein LC3 in rat. Methods: NPCs in normal nucleus pulposus tis?鄄sues of SD rats were isolated and cultured, which were then identified by toluidine blue, alixin staining and Ⅱ collagen immunofluorescence. The cells were expanded, and the second generation NPCs were cultured in medium with pH 6.2, 6.4, 6.8, 7.0, 7.2 and 7.4 for 24h. Immunofluorescence was used to observe the ex?鄄pression of OGR1 in NPCs. Western blotting was used to examine OGR1. After incubation at pH 6.4 for 0h, 3h, 6h, 12h, 24h, 48h, the expression levels of LC3-Ⅰ and LC3-Ⅱ in NPCs were detected by Western blotting. The OGR1 interference sequences of three different targets were constructed, and the most suitable titer lentivirus was detected. NPCs cells were transfected with the optimal titer lentivirus, and the silencing effects of different interference sequences were detected by Western blotting and Real-time PCR. The interfer?鄄ence series were used to construct the OGR1-shRNA lentiviral vector. The second generation NPCs were di?鄄vided into 4 groups: normal group(DMEM medium), pH 6.4 medium group(blank group), pH 6.4 medium emp?鄄ty vector group(empty vector group), pH 6.4 medium OGR1 -shRNA lentiviral transfection group(transfection group), and the expressions of LC3-Ⅱ and p62 protein were detected by Western blotting after 24 hours of culture. Alcian staining was used to observe the expression of proteoglycan in NPCs. Results: (1)The isolated cultured cells highly expressed proteoglycan and type Ⅱ collagen, which were consistent with the NPCs phe?鄄notype. (2)The expression level of OGR1 protein was correlated with the pH value of the medium. When the pH value lower than 7.0, the expression levels of OGR1 and LC3-Ⅱ protein significantly increased(P<0.05), and the expression level was the highest at pH 6.4. (3)When cultured in a medium with a pH of 6.4, the ex?鄄pression levels of LC3-Ⅰ and LC3-Ⅱ in the cells were related to the culture time, which peaked at 24h and remained high at 48h. (4)Three interfering sequences had silent effect on OGR1, which was significantly different from the control group(P<0.05), and OGR1-shRNA1 had the best silencing effect. (5)After the cells were cultured for 24 hours in the medium with pH 6.4 in the transfection group, the expression level of LC3-II was significantly lower than that in the empty group(P<0.05); the p62 expression in the transfection group was significantly higher than that in the empty vector group(P<0.05). The expression of proteoglycan in the transfection group was lower than that in the empty group(P<0.05). Conclusions: The acid environment can promote OGR1 protein expression and autophagy of NPCs in rats. OGR1 mediates the activation of au?鄄tophagy and affects the function of NP cells. |
投稿时间:2019-02-28 修订日期:2019-05-12 |
DOI:10.3969/j.issn.1004-406X.[year_id].07.641.8 |
基金项目:广西自然科学基金-面上项目(编号:2016GXNSFAA 380058) |
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