药用植物的抗增殖和凋亡活性
Antiproliferative and Apoptotic Activities of the Medicinal Plant .
作者信息
Molina-Romo Edna D, Garibay-Escobar Adriana, Valenzuela-Antelo Olivia, Ruiz-Bustos Eduardo, Martínez Javier Hernández, Vélazquez Carlos, Rascón-Valenzuela Luisa A, Robles-Zepeda Ramón Enrique
机构信息
Department of Chemical- Biological Sciences, University of Sonora, Hermosillo, Sonora, Mexico.
Unit of Support Services in Analitycal Resolution, Veracruzana University, Xico, Veracruz, Mexico.
出版信息
Pharmacognosy Res. 2018 Jan-Mar;10(1):55-59. doi: 10.4103/pr.pr_38_17.
BACKGROUND
is a spiny shrub found in Northwest Mexico desert, with traditional medicinal use to treat several diseases including cancer.
OBJECTIVE
The aims of the present study were to evaluate the antiproliferative and apoptotic activities of the aerial parts of this plant.
MATERIALS AND METHODS
The methanol extract and its fractions were prepared using several solvents. The antiproliferative activity was evaluated by the (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium) (MTT) assay on HeLa, A549, RAW 264.7, M12.C3.F6, and L-929 cell lines, and the apoptotic activity using Annexin V and (5,50,6,60-tetra-chloro-1,10,3,30-tetra-ethylbenzimidazol-carbocyanine iodide) staining. The most active fraction was further separated by column chromatography.
RESULTS
The most active fraction was hexane with an IC of 90.4 μg/mL against RAW 264.7, 94 μg/mL against M12.C3.F6, 165.5 μg/mL against HeLa and 187.7 μg/mL against A549 cell lines. In apoptotic activity assays the methanol extract and its -hexane fraction were found to induce mitochondrial depolarization in HeLa cells (83 and 87% respectively), and both induced the externalization of the phosphatidylserine increasing the percentage of cells in early apoptosis from 1.4% in untreated control cells, to 1.9% and 3.5% for methanol extract and -hexane fraction-treated cells, respectively, statistically different for the total percentage of apoptotic cells ( < 0.05).
CONCLUSIONS
These results show that has antiproliferative and apoptotic activities and confirms its use in traditional medicine.
SUMMARY
The methanol extract and its fractions using several solvents were evaluated in the antiproliferative activity by the MTT assay on HeLa, A549, RAW 264.7, M12.C3.F6, and L-929 cell lines, and the apoptotic activity using Annexin V and (5,50,6,60-tetra-chloro-1,10,3,30-tetra-ethylbenzimidazol-carbocyanine iodide) staining. The most active fraction against cell lines was hexane. In apoptotic activity assays, the methanol extract and its -hexane fraction were found to induce mitochondrial depolarization. This results we showed that has antiproliferative and apoptotic activities . DMEM: Dulbecco's modified eagle's medium, DMSO: Dimethyl sulfoxide, MTT: (3-4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium), JC-1: (5,50,6,60-tetra-chloro-1,10,3,30-tetra-ethylbenzimidazol-carbocyanine iodide), FBS: Fetal bovine serum, CAPE: Caffeic acid phenethyl ester, PBS: Phosphate-buffered saline.
背景
[植物名称]是一种生长在墨西哥西北部沙漠的多刺灌木，在传统医学中用于治疗包括癌症在内的多种疾病。
目的
本研究旨在评估该植物地上部分的抗增殖和凋亡活性。
材料与方法
使用多种溶剂制备甲醇提取物及其馏分。通过（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐）（MTT）法在HeLa、A549、RAW 264.7、M12.C3.F6和L-929细胞系上评估抗增殖活性，并使用膜联蛋白V和（5,5′,6,6′-四氯-1,1′,3,3′-四乙基苯并咪唑羰花青碘化物）染色评估凋亡活性。活性最强的馏分通过柱色谱进一步分离。
结果
活性最强的馏分是己烷，对RAW 264.7细胞系的IC50为90.4 μg/mL，对M12.C3.F6细胞系为94 μg/mL，对HeLa细胞系为165.5 μg/mL，对A549细胞系为187.7 μg/mL。在凋亡活性测定中，发现甲醇提取物及其己烷馏分可诱导HeLa细胞线粒体去极化（分别为83%和87%），且两者均诱导磷脂酰丝氨酸外化，使早期凋亡细胞百分比从未处理对照细胞的1.4%分别增加到甲醇提取物处理细胞的1.9%和己烷馏分处理细胞的3.5%，凋亡细胞总数有统计学差异（P<0.05）。
结论
这些结果表明[植物名称]具有抗增殖和凋亡活性，并证实了其在传统医学中的应用。
总结
使用多种溶剂的甲醇提取物及其馏分通过MTT法在HeLa、A549、RAW 264.7、M12.C3.F6和L-929细胞系上评估抗增殖活性，并使用膜联蛋白V和（5,5′,6,6′-四氯-1,1′,3,3′-四乙基苯并咪唑羰花青碘化物）染色评估凋亡活性。对细胞系活性最强的馏分是己烷。在凋亡活性测定中，发现甲醇提取物及其己烷馏分可诱导线粒体去极化。我们的结果表明[植物名称]具有抗增殖和凋亡活性。DMEM：杜尔贝科改良伊格尔培养基，DMSO：二甲基亚砜，MTT：（3-4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐），JC-1：（5,5′,6,6′-四氯-1,1′,3,3′-四乙基苯并咪唑羰花青碘化物），FBS：胎牛血清，CAPE：咖啡酸苯乙酯，PBS：磷酸盐缓冲盐水。
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