癌变·畸变·突变 ›› 2019, Vol. 31 ›› Issue (1): 45-48.doi: 10.3969/j.issn.1004-616x.2019.01.008

• 论著 • 上一篇    下一篇

枸杞多糖对脊髓神经细胞辐射损伤后的保护作用研究

关素珍, 德小明, 庞克华, 杨惠芳   

  1. 宁夏医科大学公共卫生与管理学院职业卫生与环境卫生学系, 宁夏 银川 750001
  • 收稿日期:2018-05-28 修回日期:2018-12-05 出版日期:2019-01-31 发布日期:2019-01-31
  • 通讯作者: 杨惠芳,E-mail:joyceyhf@163.com E-mail:joyceyhf@163.com
  • 作者简介:关素珍,E-mail:clever2066@sina.com。
  • 基金资助:
    宁夏回族自治区自然科学基金重点项目(2018AAC02007)

Protective effects of lycium barbarum polysaccharides on spinal cord neurons after radiation injury

GUAN Suzhen, DE Xiaoming, PANG Kehua, YANG Huifang   

  1. Department of Occupational Health and Environmental Health, School of Public Health and Management, Ningxia Medical University, Yinchuan 750001, Ningxia, China
  • Received:2018-05-28 Revised:2018-12-05 Online:2019-01-31 Published:2019-01-31

摘要: 目的:研究枸杞多糖(LBP)对辐射损伤脊髓神经(SCN)细胞的保护作用,并观察自噬相关蛋白LC3 Ⅱ/I表达的变化,探讨LBP对辐射损伤保护作用的机制。方法:体外培养SCN细胞,应用不同剂量(0、2、6、10 Gy)X射线辐照损伤后,采用四甲基噻唑蓝(MTT)法检测细胞存活率确定最佳辐射剂量,建立辐射损伤模型。用不同浓度(15、25、40 mg/L)LBP对辐射损伤的SCN细胞进行干预后,分别采用MTT法检测细胞存活率以确定最适LBP干预剂量;用最佳辐射剂量和LBP干预后,采用Western blot和免疫组化法检测自噬相关蛋白LC3 Ⅱ/I的表达。结果:X射线辐照后,SCN细胞存活率在2~10 Gy范围内随着照射剂量的增加而逐渐降低,和正常对照组比较差异均具有统计学意义(P < 0.05);MTT实验结果显示,与辐射组相比,不同浓度LBP干预后SCN细胞存活率均明显升高,差异具有统计学意义(P < 0.05),故确定10 Gy X射线照射和40 mg/L LBP干预进行后续造模。免疫组化和Western blot检测结果均显示,与正常对照组相比,辐射组细胞的自噬相关蛋白LC3 Ⅱ/I表达明显升高,差异具有统计学意义(P < 0.05);与辐射组相比,LBP+辐射组细胞LC3 Ⅱ/I蛋白表达亦明显升高,差异具有统计学意义(P < 0.05)。结论:LBP对体外培养的脊髓神经元辐射损伤具有保护作用,可能与LBP促进自噬相关蛋白LC3 Ⅱ/I表达有关。

关键词: 枸杞多糖, 辐射损伤, 脊髓神经细胞, 自噬, LC3 II/I

Abstract: OBJECTIVE:To investigate protective effects of lycium barbarum polysaccharides (LBP) on radiation-injured spinal cord neurons in vitro. METHODS:SCN cells from spinal cords were cultured in vitro and were injured by exposure to different doses of X-ray irradiation (0,2,6,10 Gy). MTT assay was used to detect cell viability and to determine the best radiation dose. A radiation injury model was established. Irradiated cells were exposed to different concentrations of LBP (15,25,40 mg/L) to detect its protective effects. MTT assay was used to detect the cell survival rate and to determine the optimal dose of LBP intervention. The number of autophagy lysosomes was observed by electron microscope,and expression of the LC3 Ⅱ/I of autophagy-related genes was detected by Western blot and immunohisto-chemistry. RESULTS:Radiation exposure significantly reduced the survival rate of spinal cord nerve cells as shown by data from the MTT assay (P < 0.05). At the concentration of 40 mg/L,Western blot and immunohistochemistry results show that expression of gene LC3 Ⅱ/I was increased in the LBP + radiation group,with statistical significance difference (P < 0.05). CONCLUSION:LBP protected spinal cord neurons from radiation injury in vitro which may be related to its promotion of autophagy-related protein LC3Ⅱ/I expression.

Key words: lycium barbarum polysaccharides, spinal nerve cells, radiation damage, autophagy, LC3 II/I

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