运用化学计量学法高通量筛选人参抗老年痴呆的氨基酸成分

High-throughput Screening of Ginseng for Anti-Alzheimer′s Amino Acid Composition Using Chemometrics

应用Ellman改良法和MTT法对人参水提物超滤分级后的<1 kD,1~3 kD,3~10 kD,>10 kD及总提物共5个组分进行体外乙酰胆碱脂酶(AChE)抑制活性和神经母细胞瘤细胞(SH-SY5Y)保护活性筛选,对吉林省内12个样地的人参水提物<1 kD活性组分进行氨基酸检测,获得各样品的氨基酸共有峰图谱数据,并测定各样品的AChE抑制率和SH-SY5Y细胞存活率,采用主成分分析(PCA)、偏最小二乘法(PLS)和灰色关联度分析(GRA)对所获得的氨基酸共有峰数据进行处理后,可有效区分人参的不同产地,并筛选出其中潜在抗AChE活性和SH-SY5Y细胞保护活性的氨基酸成分。试验结果显示,苏氨酸(Thr)和亮氨酸(Leu)在氨基酸共有峰中对AChE的抑制活性最佳,苏氨酸(Thr)、丙氨酸(Ala)及亮氨酸(Leu)对SH-SY5Y损伤细胞具有较好的保护作用。揭示了人参水提物<1 kD组分具有良好的AChE体外抑制活性和SH-SY5Y细胞损伤保护作用。因此,苏氨酸(Thr)和亮氨酸(Leu)可能具有潜在的抗阿尔茨海默病(AD)活性,该成分为寻找抗AD寡肽类药物提供物质基础。此研究方法为简单、高通量的从中药中筛选抗AD的活性物质提供借鉴。

Ellman modified method and MTT method were used to screen the active fraction of AChE inhibition and neuroblastoma cell(SH-SY5 Y) protection from 5 groups(<1 kD, 1-3 kD, 3-10 kD and >10 kD, and total extract) in vitro after the ultrafiltration in ginseng water extract. Amino acid analyzer was used to detect amino acid of <1 kD group in water extract of 12 sample sites in Jilin Province to get the common peak spectra data of amino acids in each sample and to determine AChE inhibitory activity and SH-SY5 Y protection of each sample. Different geographical origins of ginseng could be distinguished effectively, and potential AChE inhibition components and SH-SY5 Y cell protection components could be screened out though principal component analysis(PCA), partial least squares(PLS) and grey relational degree analysis(GRA) with the data of the total amino acid peaks being processed. The experimental results showed that threonine(Thr) and leucine(Leu) have the best inhibitory activity against AChE in the common amino acid peak, threonine(Thr), alanine(Ala) and leucine(Leu) SH-SY5 Y damaged cells have better protection. The results showed that the <1 kD group in water extract of ginseng reflected good AChE inhibition and SH-SY5 Y cell protection activity. Therefore, threonine(Thr) and leucine(Leu) may have potential anti-Alzheimer′s disease activity, which provides a material basis for the search for anti-Alzheimer′s disease oligopeptide drugs. In addition, this method also provides reference for simple and high-throughput screening of anti-Alzheimer′s disease substances from Chinese medicine.