摘要
目的探讨枸杞多糖(LBP)对小鼠海马神经元细胞系HT22细胞缺糖缺氧损伤保护作用的机制。方法采用含1 mmol/L连二亚硫酸钠的无糖DMEM培养基建立HT22细胞缺糖缺氧损伤模型,设对照组、模型组和LBP高、中、低剂量(100、50、25 mg/L)组,以CCK8法检测细胞活性,流式细胞术检测检测细胞凋亡情况;Western blotting检测Bcl-2、Bax蛋白表达情况;利用实时荧光定量PCR(RT-qPCR)检测TNF-α、NF-κB、I-κBα、Caspase-9基因转录活性。结果与对照组比较,模型组HT22细胞存活率显著降低、凋亡率显著增加(P<0.05);TNF-α、NF-κB、Caspase-9基因转录活性显著上调,I-κBα基因转录活性显著下调(P<0.05);Bax蛋白表达水平显著升高,Bcl-2的蛋白表达水平显著降低(P<0.05)。与模型组比较,LBP各剂量组细胞存活率显著上升、凋亡率显著下降(P<0.05);TNF-α、NF-κB、Caspase-9基因转录活性显著下调,I-κBα基因转录活性显著上调(P<0.05);Bax蛋白表达水平显著降低,Bcl-2的蛋白表达水平显著增高(P<0.05)。结论 LBP可能是通过下调TNF-α、NF-κB、Caspase-9基因转录活性以及Bax蛋白表达,增强I-κBα基因转录活性及Bcl-2的蛋白表达,抑制缺糖缺氧损伤诱导神经细胞凋亡的发生。
Objectives To observe the protective effects of Lycium barbarum polysaccharides(LBP) on oxygen-glucose deprived mouse hippocampal neurons. Methods Cultured mouse hippocampal neurons exposed to 1 mmol/LNa2S2O4 to establish oxygenglucose deprivation model, assigned to control group, model group and LBP group with different doses(100, 50 and 25 mg/L), the cell activity rate was measured by CCK8 assay, and the apoptosis rate was analyzed by Flow Cytometry;the protein expression of Bcl-2 and Bax was measured by Western blotting;RT-qPCR assay was used to evaluate the mRNA expression of TNF-α, NF-κB, I-κBα, Caspase-9. Results Compared with control group, HT22 cell survival rate and apoptosis rate in model group were significantly reduced(P < 0.05);TNF-α, NF-κB, caspase-9 gene transcription activity was significantly increased, I-κBα gene transcription activity was significantly decreased(P < 0.05);Bax protein expression level was significantly increased, Bcl-2 protein expression level was significantly decreased(P < 0.05). Compared with control group, LBP can significantly increase cell viability(P < 0.05);reduce the cell apoptosis rate(P < 0.05);the expression of TNF-α, NF-κB, Bax and Caspase-9 were effectively down regulated;induce an increase of I-κBα mRNA leve and Bcl-2 protein level(P < 0.05). Conclusion LBP have significantly protective effects on oxygen-glucose deprivation, such protective effects may be mediated by its action on the downstream TNF-α, NF-κB, Bax, Caspase-9, and up-regulation of Bcl-2 and I-κBα.
作者
马琳
张蓓
胡秀娟
左娣
和祯泉
张春
牛建国
MA Lin;ZHANG Bei;HU Xiujuan;ZUO Di;HE Zhengjuan;ZHANG Chun;NIU Jianguo(Ningxia Key Laboratory of Cerebrocranial Diseases,Ningxia Medical University,Yinchuan 750004,China;942 Hospital of People's Liberation Army,Yinchuan 750004,China;Genneral Hospital of Ningxia Medical University,Yinchuan 750004,China)
出处
《药物评价研究》
CAS
2020年第4期683-687,共5页
Drug Evaluation Research
基金
宁夏自治区“十三五”重大科技项目(2016BZ07)
宁夏自然科学基金项目(NZ13073)
宁夏高等学校科学项目(NGY2015083)
宁夏医科大学校级项目(XY201713)。
关键词
枸杞多糖
缺糖缺氧损伤
凋亡
神经保护
Lycium barbarum polysaccharides
oxygen-glucose deprivation
apoptosis
neuroprotection
