百草枯诱导的SH-SY5Y细胞中核连蛋白2基因的表达变化

发布时间：2018-05-27 10:41

本文选题：核连蛋白(NUCB) + PQ　；参考：《中国老年学杂志》2017年13期

【摘要】：目的检测核连蛋白(NUCB)2在百草枯(PQ)诱导的SH-SY5Y细胞中的表达变化,分析NUCB2在PQ引起的SH-SY5Y细胞凋亡中的作用机制。方法采用MTT法进行SH-SY5Y细胞毒性分析。选择PQ作用的最佳浓度和时间点,进一步收取经PQ处理后的SH-SY5Y总RNA,首先采用RT-PCR方法观察NUCB2的表达趋势,然后采用real-time PCR方法进行NUCB2基因进行表达变化检测,采用ΔΔCt法进行定量比较分析。结果 MTT方法在酶标仪490 nm波长下,分析得到PQ的最佳作用浓度和时间是0.5 mmol/L作用24 h,此时细胞活力较对照组低约41.67%;采用ΔΔCt法进行定量比较分析,发现NUCB2在mRNA水平的表达量明显降低(P=0.001 6);Western印迹检测发现SH-SY5Y细胞经PQ诱导后引起了caspase-3酶原形式的表达量降低(P0.05)。结论 NUCB2基因在经PQ诱导后的SH-SY5Y细胞中表达量明显降低,分析其可能在神经细胞的凋亡机制中发挥重要作用。
[Abstract]:Objective to detect the expression of Nu CB2 in SH-SY5Y cells induced by paraquat (PQ) and to analyze the mechanism of NUCB2 in PQ induced apoptosis of SH-SY5Y cells. Methods MTT assay was used to analyze the cytotoxicity of SH-SY5Y. The optimal concentration and time point of PQ action were selected to collect the total SH-SY5Y after PQ treatment. The expression trend of NUCB2 was observed by RT-PCR method, and then the expression change of NUCB2 gene was detected by real-time PCR method. 螖 Ct method was used for quantitative comparative analysis. Results under the wavelength of 490nm, the optimal concentration and time of MTT was 0.5 mmol/L for 24 h, and the cell viability was about 41.67% lower than that of the control group, and the 螖 Ct method was used for quantitative analysis. It was found that the expression of NUCB2 in mRNA level was significantly decreased. Western blot analysis showed that the expression of caspase-3 prozyme in SH-SY5Y cells induced by PQ was decreased by P0. 05%. Conclusion the expression of NUCB2 gene in SH-SY5Y cells induced by PQ was significantly decreased, which may play an important role in the mechanism of neuronal apoptosis.
【作者单位】：河北北方学院法医教研室;河北北方学院;中国医科大学法医学院法医血清教研室;
【基金】：河北省教育厅青年指导项目(Z2017027)
【分类号】：R742.5

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