内毒素诱导退变人椎间盘髓核细胞凋亡

发布时间：2018-01-18 12:03

本文关键词：内毒素诱导退变人椎间盘髓核细胞凋亡　出处：《基础医学与临床》2017年05期 　论文类型：期刊论文

【摘要】：目的观察内毒素(LPS)刺激退变人椎间盘髓核细胞前后,核因子kapp B(NF-κB)的活化与细胞凋亡的相互关系。方法体外培养退变的人椎间盘髓核细胞,采用CCK-8法测定LPS在不同浓度下(100、200、500和1 000μg/m L)对髓核细胞增殖影响,并筛选合适的刺激浓度;设立空白对照组,单纯LPS(500μg/m L)刺激组,NF-κB抑制剂(BAY11-7082)+LPS(500μg/m L)刺激组,以Hochest33258染色以及Annexin V-PI双标法检测细胞凋亡;Western blot检测凋亡相关蛋白caspase-3、PARP、NF-κB结合蛋白P65以及磷酸化P-P65的表达。结果 CCK-8实验结果显示当LPS浓度为500μg/m L能明显降低细胞的活力。与空白对照组相比,单纯LPS刺激组细胞凋亡率上升(P0.05),P-P65表达明显增加,其凋亡蛋白活化型caspase-3和PARP降解产物表达也增高(P0.05)。而与单纯LPS刺激组相比,NF-κB抑制剂+LPS刺激组细胞凋亡率下降(P0.05),P-P65表达明显降低,其凋亡蛋白活化型caspase-3和PARP降解产物表达也降低(P0.05)。结论 NF-κB信号通路可能与椎间盘退变时髓核细胞凋亡的发生密切相关,值得进一步研究。
[Abstract]:Objective to observe the effect of endotoxin (LPS) stimulation on degenerative human disc nucleus pulposus cells. The relationship between the activation of nuclear factor kapp BnNF- 魏 B and apoptosis. Methods the degenerative human intervertebral disc nucleus pulposus cells were cultured in vitro. The effects of LPS on the proliferation of nucleus pulposus cells at different concentrations were determined by CCK-8 assay. The control group was treated with LPS(500 渭 g / mL alone, and the NF- 魏 B inhibitor BAY11-7082 (LPS(500 渭 g / mL) was used. Apoptosis was detected by Hochest33258 staining and Annexin V-PI double labeling method. The apoptosis-related protein caspase-3 was detected by Western blot. Expression of NF- 魏 B binding protein p65 and phosphorylated P-P65. Results CCK-8 results showed that when the concentration of LPS was 500 渭 g / m, L can significantly reduce the cell viability, compared with the blank control group. The expression of P-P65 was significantly increased in LPS stimulated group (P 0.05). The expression of apoptosis-activated caspase-3 and PARP degradation products was also higher than that of LPS alone. The apoptotic rate of NF- 魏 B inhibitor LPS was decreased and the expression of P-P65 was significantly decreased. The expression of apoptosis-activated caspase-3 and PARP degradation products also decreased (P 0.05). Conclusion NF- 魏 B signaling pathway may be closely related to the apoptosis of nucleus pulposus cells during disc degeneration. It is worth further study.
【作者单位】：重庆医科大学附属第一医院骨科;重庆市合川区人民医院骨科;
【基金】：国家自然科学基金(81572202)
【分类号】：R681.5
【正文快照】： 椎间盘退变是腰椎间盘突出、腰椎滑脱、腰椎管狭窄及椎间盘源性腰痛等脊柱退行性疾病的最主要原因[1],但其退变的原因和分子机制目前尚未完全阐明。核转录因子kappa B(nuclear facor-kappa B,NF-κB)是一种多功能的转录因子,其存在于大多数细胞内,在参与炎性反应及细胞损伤方

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