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