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王立侠,韩金芬.枸杞多糖与 UCA1 siRNA 单用或合用对 MPP+ 诱导帕金森病细胞模型的干预作用及机制研究[J].中国比较医学杂志,2021,31(11):55~61.
枸杞多糖与 UCA1 siRNA 单用或合用对 MPP+ 诱导帕金森病细胞模型的干预作用及机制研究
Mechanism of Lycium barbarum polysaccharide and UCA1 siRNA single or combined treatment in the MPP+ -induced Parkinson’s disease cell model
投稿时间:2020-12-01  
DOI:10. 3969 / j.issn.1671-7856. 2021. 11. 009
中文关键词:  帕金森  枸杞多糖  长链非编码 RNA UCA1  凋亡  氧化应激  细胞模型
英文关键词:Parkinson ’ s disease  Lycium barbarum polysaccharide  long-chain non-coding RNA UCA1  apoptosis  oxidative stress  cell model
基金项目:
作者单位E-mail
王立侠 郑州市第七人民医院神经电生理室,郑州 450000 itj7wb@ 163.com 
韩金芬 新乡医学院第三附属医院儿内科,河南 新乡 453003  
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中文摘要:
       目的 探讨枸杞多糖与 UCA1 siRNA 单独或联合应用对 MPP+诱导的 SH-SY5Y 细胞增殖、凋亡的影响及机制。 方法 使用 MPP+(终浓度为 0. 25、0. 5、1、2 mmol / L 的 MPP+ 处理细胞 24 h)、枸杞多糖(50、100、200、400 μg / mL 的枸杞多糖预处理细胞 1 h 后加 MPP+继续处理 24 h)处理 SH-SY5Y 细胞,选择最佳 MPP+(1 mmol / L) 及枸杞多糖作用浓度(400 μg / mL)开展后续实验。实验分为对照组(细胞不经特殊处理)、MPP+组(1 mmol / L 的 MPP+处理细胞 24 h)、枸杞多糖组(LBP 组)(400 μg / mL 的枸杞多糖预处理细胞 1 h 后加 1 mmol / L 的 MPP+继续处理 24 h)、si-UCA1 组(1 mmol / L 的 MPP+处理细胞后转染 UCA1 siRNA)、枸杞多糖+si-UCA1 组。 MTT 法检测细胞活力;流式细胞术检测细胞凋亡率、ROS 及膜电位水平。 结果 与对照组比较,MPP+组细胞存活率、膜电位显著降低,细胞凋亡率、ROS 水平显著升高(P<0. 05)。 与 MPP+组比较,LBP 组和 si-UCA1 组细胞存活率、膜电位显著升高,细胞凋亡率、ROS 水平显著降低(P<0. 05)。 与 LBP 组或 si-UCA1 组比较,LBP+si-UCA1 组细胞存活率、膜电位显著升高,细胞凋亡率、ROS 水平显著降低(P<0. 05)。 结论 枸杞多糖及抑制 UCA1 均可促进 MPP+诱导的 SH- SY5Y 细胞增殖,抑制凋亡,两者联合对细胞增殖、凋亡影响更显著,机制可能与抑制线粒体凋亡途径有关。
英文摘要:
       Objective To investigate the effect of Lycium barbarum polysaccharide and UCA1 siRNA individually, or in combination, on the proliferation and apoptosis of SH-SY5Y cells induced by MPP+ and its mechanism. Methods SH-SY5Y cells were treated with MPP+(0. 25, 0. 5, 1 and 2 mmol / L for 24 hours) and LBP (50, 100, 200 and 400 μg / mL Lycium barbarum polysaccharide pretreatment for 1 hour and then treated with MPP+ for 24 hours). The optimal concentrations of MPP+ ( 1 mmol / L) and LBP ( 400 μg / mL) were selected for subsequent experiments. The experiment was divided into four groups: control group ( without treatment ), MPP+ group ( cells were treated with 1 mmol / L MPP+ for 24 hours), Lycium barbarum polysaccharide group ( cells were pretreated with 400 μg / mL Lycium barbarum polysaccharide for 1 hour and then treated with 1 mmol / L MPP+ for 24 hours), si-UCA1 group ( cells were treated with 1 mmol / L MPP+ and then transfected with si-UCA1) and barbarum polysaccharide + si-UCA1 group. MTT assays were used to assess cell viability and flow cytometry was used to measure the apoptosis rate, ROS level and membrane potential. Results Compared with the control group, the cell survival rate and membrane potential in the MPP+ group were reduced significantly and the apoptosis rate and ROS level were increased significantly (P<0. 05). Compared with the MPP+ group, the cell survival rate and membrane potential in Lycium barbarum polysaccharide and si-UCA1 groups were increased significantly and the apoptosis rate and ROS level were decreased significantly (P<0. 05). Compared with Lycium barbarum polysaccharide and si-UCA1 groups, the cell survival rate and membrane potential in the LBP+si-UCA1 group were increased significantly and the apoptosis rate and ROS level were decreased significantly ( P< 0. 05 ). Conclusions Lycium barbarum polysaccharide and inhibition of UCA1 expression promote the proliferation of SH-SY5Y cells induced by MPP+ and inhibit apoptosis. Their combination has a more obvious effect on cell proliferation and apoptosis. The mechanism may be related to inhibition of mitochondrial apoptosis.
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