Introduction

Sirtuin1 (SIRT1), one of NAD⁺-dependent protein deacetylases, is proved to be neuroprotective in aging diseases, but its effect on neuronal apoptosis has not been clarified. To investigate the role of SIRT1 in inhibiting neuronal apoptosis, SIRT1 was interfered or overexpressed in cortical neurons.

Methods

We exerted overloading laminar shear stress with 10 dyn/cm² for 4, 8, and 12 h on neurons to cause cortical neuronal apoptosis, and the apoptosis percentage was tested by TUNEL assay. The adenovirus plasmids containing SIRT1 RNA interference or SIRT1 wild type gene were transfected into neurons before shear stress loading. SIRT1 mRNA and protein level were tested by Real-time PCR, immunofluorescence and western blots assay.

Results

SIRT1 was primarily expressed in nucleus of cortical neurons, and its mRNA level was significantly increased after 4 h stimulation. SIRT1 RNAi cortical neurons had higher TUNEL positive cells, while SIRT1 overexpression significantly decreased the percentage of died cells induced by shear stress compared to control group.

Conclusions

SIRT1 plays a neuroprotective role in shear stress induced apoptosis and could be as potential pharmacological targets against neuronal degeneration in future.

中文翻译：

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SIRT1 抑制大鼠皮质神经元中高剪切应力诱导的细胞凋亡

介绍

Sirtuin1 (SIRT1) 是一种 NAD ⁺依赖性蛋白脱乙酰酶，已被证明在衰老疾病中具有神经保护作用，但其对神经元凋亡的影响尚未阐明。为了研究 SIRT1 在抑制神经元凋亡中的作用，SIRT1 在皮质神经元中受到干扰或过表达。

方法

对神经元施加10 dyn/cm ²的超负荷层流剪切应力4、8、12 h，引起皮层神经元凋亡，采用TUNEL法检测凋亡百分比。在剪切应力加载之前，将含有 SIRT1 RNA 干扰或 SIRT1 野生型基因的腺病毒质粒转染到神经元中。通过实时荧光定量PCR、免疫荧光和蛋白质印迹测定检测SIRT1 mRNA和蛋白质水平。

结果

SIRT1主要在皮层神经元细胞核中表达，刺激4 h后其mRNA水平显着升高。与对照组相比，SIRT1 RNAi 皮质神经元具有较高的 TUNEL 阳性细胞，而 SIRT1 过表达显着降低了剪切应力诱导的死亡细胞百分比。

结论

SIRT1在剪切应力诱导的细胞凋亡中发挥神经保护作用，未来可能成为对抗神经元变性的潜在药理学靶点。