锂剂促进自噬保护受损神经细胞的研究

Study of the neural protective effect of lithium on enhancement of autophagy in vitro

目的:研究锂剂能否通过促进细胞自噬发挥神经细胞保护作用。方法:将SH-SY5Y神经细胞分为4组,包括对照组(正常换液),模型组(培养液含有200μmol/L H2O2的培养液),锂剂组(培养液含200μmol/L H2O2及1.0 mmol/L Li Cl的培养液),3-MA干预组(培养液含200μmol/L H2O2、1.0 mmol/L LiCl及5 mmol/L 3-MA的培养液)。培养6 h后进行MTT检测及Beclin 1、LC3b免疫组化染色,评估细胞存活情况及自噬水平。结果:3-MA干预组细胞存活率较模型组降低(P<0.05),锂剂组细胞存活率较模型组提高(P<0.05)。经过3-MA干预后,细胞存活率较对照组、模型组及锂剂组均有所降低(P<0.05)。经过H2O2处理后,细胞Beclin 1、LC3b染色面积增大,染色加深。经锂剂处理后细胞Beclin 1、LC3b染色面积进一步增大,染色更深。3-MA干预组细胞Beclin 1、LC3b染色面积较对照组大、染色深,但细胞染色面积较模型组和锂剂组均小,且染色更为浅淡。结论:锂剂能够促进受损神经细胞存活,促进自噬作用是锂剂发挥神经保护作用的机制之一。

Objective:To investigate whether lithium can exert neuroprotective effects by promoting autophagy.Methods:SH-SY5Y cells were divided into 4 groups including control group(handled with normal culture solution),model group(handled with 200μmol/L H2O2),lithium group(handled with 200μmol/L H2O2 and 1.0 mmol/L LiCl medium),3-MA group(handled with 200μmol/L H2O2,1.0 mmol/L LiCl and 5 mmol/L 3-MA).After 6 hours of culture,MTT assay and immunohistochemical staining of Beclin 1 and LC3b were performed to evaluate cell survival and autophagy.Results:The cell survival rate of lithium group was significantly high than that of the model group(P<0.05),while the 3-MA group was lower(P<0.05).After 3-MA intervention,the cell survival rate was lower than that of control group,model group and lithium group(P<0.05).After H2O2 treatment,the staining area of Beclin 1 and LC3b was increased and the staining was deeper,and after LiCl handling,the staining area of Beclin 1 and LC3b was further increased and the staining was more deeper.The staining area of Beclin 1 and LC3b in 3-MA group was larger than that in control group,but was smaller than that in model group and lithium group,and the staining was lighter.Conclusion:Lithium can promote the survival of damaged nerve cells,and inducing autophagy is probably one of the neuroprotective mechanisms of lithium.