摘要
利用不同方法将梅花鹿鹿茸提取物(Pilose anlter extracts,PAEs)分成不同组分,用1-甲基-4-苯基吡啶离子(MPP^(+))诱导的PC12细胞损伤模型,筛选并确定PAEs具有神经保护作用的活性组分。提取PAEs,利用超滤离心、热处理的方法对PAEs进行处理,获得PAEs、PAEsH(热处理)、PAEs_(30)(超滤离心分子质量>30 kDa组分),分别观察其对MPP^(+)诱导的PC12细胞损伤保护作用,并进一步追踪分离。选择活性最强组分PAEs_(30)进一步加热、酶解处理后得到PAEs_(30-H)(热处理)与PAEs_(30-E)(酶解),分别观察其对MPP^(+)诱导的PC12细胞损伤保护作用,并进一步追踪分离,最终确定有效活性组分。结果表明:与MPP^(+)损伤的PC12细胞模型相比,质量浓度>0.5 mg/mL的PAEs、PAEs_(30)可显著抑制MPP^(+)对PC12细胞的损伤,其中PAEs_(30)组分活性最强。对PAEs_(30)进行热处理及酶解处理后得到的各组分对MPP^(+)损伤的PC12细胞均有明显的神经保护作用,质量浓度≥0.125 mg/mL均可显著提高细胞的相对生存率。可见,PAEs中具有MPP^(+)诱导神经细胞损伤修复作用的活性成分,初步鉴定为对热稳定、分子质量为10 kDa或35 kDa的蛋白类成分,这为后续的质量标准制定及提取工艺提供理论依据。
The sika deer pilose anlter extracts(PAEs)were divided into different components by different methods.The PC12 cell injury model induced by 1-methyl-4-phenylpyridine ion(MPP^(+))was screened to determine the neuroprotective effects of PAEs.The active components lay the foundation for the next step of new drug development.PAEs were extracted by a patented method in our laboratory,and treated separately by ultrafiltration centrifugation and heat treatment to obtain different components of PAEs,PAEsH(heat treatment),and PAEs_(30)(ultrafiltration centrifugal molecular weight>30 kDa components).Their protective effects on MPP^(+)-induced PC12 cell damage were observed,and separation was further tracked.The most active component PAEs_(30) was selected for further heat treatment and enzymatic hydrolysis to obtain PAEs_(30-H)(heat treatment)and PAEs_(30-E)(enzymatic hydrolysis).Their protective effects on MPP^(+)-induced PC12 cell damage were observed,and tracking of separation was further carried out to determine the effective active component.Compared with the MPP^(+)injured PC12 cell model,PAEs,and PAEs_(30) at concentrations above 0.5 mg/mL could significantly inhibit MPP^(+)damage of PC12 cells,with PAEs_(30) component having the strongest activity.After heat treatment and enzymolysis treatment of PAEs_(30)(PAEs_(30-H) and PAEs_(30-E)),each component had a significant neuroprotective effect on MPP^(+)injured PC12 cells,and concentrations of 0.125 mg/mL and above could significantly improve cell survival rate.PAEs repaired the damage of MPP^(+)-induced nerve cell.The main active components were initially identified as protein components with thermal stability and molecular weight of 10 kDa or 35 kDa.The research provides experimental data for subsequent quality standard formulation and extraction process.
作者
赵鑫
陈思
李超华
刘洋
孙娅楠
王毅
何忠梅
ZHAO Xin;CHEN Si;LI Chaohua;LIU Yang;SUN Yanan;WANG Yi**;HE Zhongmei(College of Chinese Medicinal Materials,Jilin Agricultural University,Changchun 130118,China;Experimental Research Center,China Academy of Chinese Medical Science,Beijing 100700,China;College of Bioengineering and Food Science,Hubei University of Technology,Wuhan 430068,China)
出处
《吉林农业大学学报》
CSCD
北大核心
2024年第6期1040-1048,共9页
Journal of Jilin Agricultural University
基金
国家自然科学基金面上项目(81603285)
北京自然科学青年基金面上项目(7184230,7192141)
中央级公益性科研院所基本科研业务费专项(ZZ13-YQ-077)。
关键词
鹿茸提取物
活性追踪分离
神经细胞损伤修复
超滤离心
pilose anlter extract
activity tracking separation
repairation of neuronal damage
ultrafiltration centrifugation
