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应用显微镜和光谱学研究牛膝草叶的抗癌潜力。

Application of microscopy and spectroscopy in investigating anti-cancer potential of Achyranthes aspera L. leaves.

机构信息

Department of Biological Sciences, University of Veterinary and Animal Sciences, Lahore, Pakistan.

Department of Botany, Govt. Queen Mary Graduate College, Lahore, Pakistan.

出版信息

Microsc Res Tech. 2024 May;87(5):1031-1043. doi: 10.1002/jemt.24495. Epub 2024 Jan 11.

Abstract

The genus Achyranthes belong to the family Amaranthaceae which constitutes an important group of herbs and shrubs with immense medicinal value. The present research work was conducted to investigate the anticancer potential of Achyranthes aspera L. leaves by focusing on the antioxidant, aniproliferative and antimitotic activities of leaf extracts. Plant extraction was carried out by soxhelt method with different solvents. Phytochemical characterization of the plants extracts using chemical methods identified the presence of cardiac glycosides, saponins, coumarins, proteins, tannins, flavonoids and triterpenes. Alkaloid was present in methanolic and ethanolic extract. High performance liquid chromatography showed presence of different concentration of myricetin, quercetin and kaempferol in different extracts with the highest concentration of myricetin (84.53 μg/mL) in n-butanolic extract. The extracts were then tested for antioxidant activity using 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging assay by spectrophotometric method. In DPPH radical scavenging assay, antioxidant activity of A. aspera ranged between 79.78 ± 0.034% and 58.63 ± 0.069%. Highest antioxidant activity was observed for methanolic extract and lowest for acetone. Antimitotic activity was determined by using Allium cepa assay in which microscopic investigation was carried out to observe normal and abnormal phases of mitosis. In this assay, n-butanolic extract had highest antimitotic activity with minimum mitotic index at 2 mg/mL (57 ± 0.0351%). The plant extracts also caused chromosomal and mitotic aberrations which were clearly observed under 40× and 100× magnification of compound microscope. Antiproliferative activity was determined by using yeast cell model in which light microscope with hemocytometer was used for cell counting. In case of Antiproliferative activity, the ethyl acetate extract of A. aspera had highest antiproliferative activity with lowest cell viability (22.14 ± 0.076%) at highest extract concentration (2 mg/mL) while methanol extract of A. aspera had highest antiproliferative activity with lower cell viability (24.24 ± 0.057%) at lowest extract concentration (0.25 mg/mL). The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent. RESEARCH HIGHLIGHTS: Achyranthes aspera L. leaves have various phytochemicals which contribute to its medicinal properties Various extracts of the leaves of A. aspera L. possess antioxidant, antimitotic and antiproliferative potential The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent.

摘要

牛膝属属于苋科,是一类具有重要药用价值的草本和灌木植物。本研究旨在探讨牛膝的抗癌潜力,重点研究其叶片提取物的抗氧化、抗增殖和抗有丝分裂活性。采用索氏提取法,用不同溶剂对植物进行提取。利用化学方法对植物提取物进行植物化学鉴定,确定存在强心苷、皂苷、香豆素、蛋白质、单宁、类黄酮和三萜。甲醇和乙醇提取物中含有生物碱。高效液相色谱法显示,不同提取物中存在不同浓度的杨梅素、槲皮素和山奈酚,其中正丁醇提取物中杨梅素的浓度最高(84.53μg/mL)。然后通过分光光度法测定 2,2-二苯基-1-苦基肼(DPPH)自由基清除试验,测试提取物的抗氧化活性。在 DPPH 自由基清除试验中,牛膝的抗氧化活性在 79.78±0.034%至 58.63±0.069%之间。甲醇提取物的抗氧化活性最高,丙酮提取物的抗氧化活性最低。采用洋葱大蒜鳞茎实验测定抗有丝分裂活性,通过显微镜观察有丝分裂的正常和异常阶段。在该实验中,正丁醇提取物的抗有丝分裂活性最高,在 2mg/mL 时最低有丝分裂指数为 57±0.0351%。植物提取物还导致染色体和有丝分裂异常,在 40×和 100×的复式显微镜下可以清楚地观察到这些异常。通过酵母细胞模型测定细胞增殖抑制活性,使用带血细胞计数器的光学显微镜进行细胞计数。在细胞增殖抑制活性方面,牛膝的乙酸乙酯提取物在最高提取物浓度(2mg/mL)时具有最高的增殖抑制活性和最低的细胞活力(22.14±0.076%),而牛膝的甲醇提取物在最低提取物浓度(0.25mg/mL)时具有最高的增殖抑制活性和较低的细胞活力(24.24±0.057%)。研究结果表明,牛膝叶片提取物具有作为抗癌药物来源的巨大潜力。研究亮点:牛膝属植物含有多种具有药用特性的植物化学物质。牛膝属植物叶片的各种提取物具有抗氧化、抗有丝分裂和抗增殖的潜力。本研究结果表明,牛膝叶片提取物具有作为抗癌药物来源的巨大潜力。

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