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鹿茸多肽通过 miR-613-HDAC6 通路对阿尔茨海默病细胞模型的影响。

Effects of velvet antler polypeptides on Alzheimer's disease cell model via miR-613 HDAC6 pathway.

机构信息

College of Integration of Chinese and Western Medicine, Hebei University of Chinese Medicine, Shijiazhuang, China.

College of Pharmacy, Henan University of Traditional Chinese Medicine, Zhengzhou, China.

出版信息

Pak J Pharm Sci. 2020 May;33(3(Special)):1427-1433.

Abstract

To study the effect of velvet antler polypeptides (VAP) on Alzheimer's disease (AD) cell model, Aβ was used to induce SK-N-SH cells to obtain AD cell model. The MDA, SOD, GSH-Px levels were determined using relevant kits. Flow cytometry was conducted to detect apoptosis, Western blot was employed to measure Bcl-2, Bax, HDAC6 protein expression, and qPCR was used to assay microRNA (miR)-613 and HDAC6 mRNA levels. Target Scan prediction combined with dual luciferase reporting experiments was conducted to analyze the targeting relationship between miR-613 and HDAC6. miR-613 was transfected in SK-N-SH cells; Alternatively, anti-miR-613 was transfected, followed by Aβ and 80 mg/L of VAP. The AD model cells showed increased MDA content, apoptosis rate, Bax protein expression, HDAC6 mRNA and protein expression, but lower SOD, GSH-Px activities, Bcl-2 protein level, and miR-613 expression (p<0.05). VAP reduced MDA content, apoptosis rate, Bax protein expression, HDAC6 mRNA and protein expression, but enhanced SOD, GSH-Px activities, Bcl-2 protein level, and miR-613 expression (p<0.05). Over-expression of miR-613 increased SOD, GSH-Px activities, and Bcl-2 protein expression in AD model cells, but reduced HDAC6 protein levels, MDA content, apoptosis rate, and Bax protein levels (p<0.05). VAP may regulate Aβ-induced apoptosis so as to treat Alzheimer's disease.

摘要

为研究鹿茸多肽(VAP)对阿尔茨海默病(AD)细胞模型的影响,采用β淀粉样蛋白(Aβ)诱导 SK-N-SH 细胞构建 AD 细胞模型。采用相关试剂盒测定 MDA、SOD、GSH-Px 水平,流式细胞术检测细胞凋亡,Western blot 检测 Bcl-2、Bax、HDAC6 蛋白表达,qPCR 检测 microRNA(miR)-613 和 HDAC6 mRNA 水平。采用靶基因预测软件 Target Scan 结合双荧光素酶报告实验分析 miR-613 与 HDAC6 的靶向关系。转染 SK-N-SH 细胞 miR-613;或转染 anti-miR-613,再加入 Aβ和 80mg/L 的 VAP。AD 模型细胞 MDA 含量、凋亡率、Bax 蛋白表达、HDAC6 mRNA 和蛋白表达增加,SOD、GSH-Px 活性、Bcl-2 蛋白水平、miR-613 表达降低(p<0.05)。VAP 降低 MDA 含量、凋亡率、Bax 蛋白表达、HDAC6 mRNA 和蛋白表达,提高 SOD、GSH-Px 活性、Bcl-2 蛋白水平、miR-613 表达(p<0.05)。过表达 miR-613 增加 AD 模型细胞 SOD、GSH-Px 活性、Bcl-2 蛋白表达,降低 HDAC6 蛋白水平、MDA 含量、凋亡率、Bax 蛋白水平(p<0.05)。VAP 可能通过调节 Aβ 诱导的细胞凋亡发挥治疗 AD 的作用。

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