LCMS 法测定及对 L6 和 SK-N-MC 细胞系的细胞毒性。

LCMS Determination and Cytotoxicity of on L6 and SK-N-MC Cell Lines.

机构信息

Department of Pharmacy, Immunology Laboratory, Annamalai University, Annamalai Nagar, 608002, Tamilnadu, India.

Centre for Cell Biology and Drug Discovery, Faculty of Engineering and Technology, Annamalai University, Annamalai Nagar, 608002, Tamilnadu, India.

出版信息

Anticancer Agents Med Chem. 2023;23(12):1376-1387. doi: 10.2174/1871520623666230320144607.

DOI:10.2174/1871520623666230320144607

PMID:36959155

Abstract

OBJECTIVE

The present study aimed to investigate the cytotoxic effect of various extracts derived from Linn. leaves on rat L6 and human SK-N-MC neuroblastoma cell lines and determine the secondary metabolites responsible for the cytotoxicity of .

METHODS

Successive solvent extraction of leaves was carried out using the Soxhlet apparatus with solvents such as petroleum ether, chloroform, ethyl acetate, and ethanol. HPTLC fingerprinting and LC-MS studies were performed to assess the presence of secondary metabolites, such as flavonoids and phenols, in the ethyl acetate extract. Furthermore, the cytotoxic effect of extracts was tested on rat skeletal muscle cell line L6 and human neuroblastoma cell line SK-N-MC using MTT assay.

RESULTS

The total phenolic content of ethyl acetate and ethanol extracts of were 72.67 and 60.73 mg, respectively, of GAE/g dry weight of the extract. The total flavonoid content of ethyl acetate and ethanol extract of were 107.33 and 40.66 mg of Quercetin equivalents/g dry weight of the extract. LCMS analysis demonstrated that the flavonoids in specific Naringenin, Diosmetin, Glycitin, and Genistein might play a prominent role in the cytotoxicity of . The cytotoxicity study revealed that the extracts of were non-toxic to rat L6 myotubes, and the IC values of the various extracts, such as APPE, APCH, APEA, and APET, were >100 μg/ml. The extracts exhibited cytotoxic activity against human neuroblastoma SK-N-MC cells, and the IC values of APPE, APCH, APEA, APET, and the standard drug "Cisplatin" were >100, >100, 64.88, >100, and 3.72 μg/ml, respectively.

CONCLUSION

It was concluded from the study that the extracts of were cytotoxic to neuroblastoma cell lines but non-toxic to normal cell lines. HPTLC and LC-MS studies confirmed that flavonoids in the ethyl acetate extract could be responsible for the biological activity.

摘要

目的

本研究旨在探讨 Linn. 叶片的不同提取物对大鼠 L6 和人 SK-N-MC 神经母细胞瘤细胞系的细胞毒性作用，并确定导致毒性的次生代谢物。

方法

采用索氏提取器用石油醚、氯仿、乙酸乙酯和乙醇等溶剂对 Linn. 叶片进行连续溶剂提取。采用 HPTLC 指纹图谱和 LC-MS 研究评估乙酸乙酯提取物中黄酮类和酚类等次生代谢物的存在。此外，采用 MTT 测定法检测提取物对大鼠骨骼肌细胞系 L6 和人神经母细胞瘤细胞系 SK-N-MC 的细胞毒性作用。

结果

Linn. 叶片的乙酸乙酯和乙醇提取物的总酚含量分别为 72.67 和 60.73 mg GAE/g 提取物干重。乙酸乙酯和乙醇提取物中 Linn. 的总黄酮含量分别为 107.33 和 40.66 mg 槲皮素当量/g 提取物干重。LCMS 分析表明，特定的柚皮苷、香叶木素、大豆苷元和染料木黄酮类化合物可能在 Linn. 的细胞毒性中发挥重要作用。细胞毒性研究表明， Linn. 提取物对大鼠 L6 肌管无毒，各种提取物（如 APPE、APCH、APEA 和 APET）的 IC 值均>100μg/ml。提取物对人神经母细胞瘤 SK-N-MC 细胞表现出细胞毒性活性，APPE、APCH、APEA、APET 和标准药物“顺铂”的 IC 值分别为>100、>100、64.88、>100 和 3.72μg/ml。