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Effects of Periplaneta americana Extract YS-F on the Proliferation and Apoptosis of Human Non-small Cell
Lung Cancer A549 Cells
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NI Lianli ，YAN Shuang ，XIAO Huai ，WU Xiumei ，HE Miao ，LI Yue （1. Key Lab of Entomological
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Biopharmaceutical R&D，Dali University，Yunnan Dali 671000，China；2. National-local Joint Engineering
Research Center of Entomoceutics，Dali University，Yunnan Dali 671000，China）
ABSTRACT OBJECTIVE：To investigate the effects of Periplaneta americana extract on the proliferation and apoptosis of
human non-small cell lung cancer A549 cells as well as its possible mechanism. METHODS：The dry bodies of P. americana were
soaked with 90％ ethanol and eluted with gradient water-methanol by polyamide column chromatography. The 20％，30％，40％，
50％，60％，70％，80％，90％ methanol elution sites（YS-A-H）were obtained. MTT method was used to screen the active site，
and the inhibition rate of different doses of active site was detected. Flow cytometry was adopted to detect cell apoptosis，cell cycle
and mitochondrial membrane potential of cells after treated with different doses of active site. RESULTS： Half inhibition
concentrations of YS-A-H were（95.25±8.42），（129.93±7.24），（221.28±12.68），（275.39±14.87），（276.76±16.32），（31.90±
5.34），（163.15±6.97），（122.81±8.36）μg/mL，respectively. YS-F had the strongest activity. After treated with 3，9，27，81 μg/mL
YS-F for 24，48，72 h，cell proliferation inhibitory rate was increased significantly at different time points；after treated for 48，72
h，that was significantly higher than same group after treated for 24 h；after 72 h treatment，that was significantly higher than same
group after 48 h treatment（P＜0.01）. There was no significant effect of 24 h treatment of 3 μg/mL YS-F and 72 h treatment of 9
μg/mL YS-F on the percentage of cells in the late stage of necrosis，24 h treatment of 3 μg/mL YS-F on the percentage of cells in
G2/M phase and 48 h treatment of 3 μ g/mL YS-F on the reduction rate of mitochondrial membrane potential （P＞0.05）. The
percentage of cells in the early stage of apoptosis，the late stage of apoptosis and the early stage of necrosis，the late stage of
necrosis，as well as the percentage of cells in the Sub-G0/G1 and S phase at each time point were significantly increased in other
different doses groups，while the percentage of cells in G0/G1 and G2/M phase was decreased significantly（P＜0.01）. In each dose
group，the percentage of cells in the early stage of apoptosis，the late stage of apoptosis and the early stage of necrosis，the late
stage of necrosis（except for the percentage of cells in the late stage of necrosis treated with YS-F 9 μg/mL for 72 h）and the
percentage of cells in Sub-G0/G1 phase，G2/M phase（except for YS-F 27，81 μg/mL for 48 h）after treated for 48，72 h were
significantly higher than same group after 24 h of treatment；the percentage of cells in G0/G1 phase，S phase and G2/M phase
（except for YS-F 9 μg/mL for 48 h）after treated for 48，72 h were significantly lower than same group after 24 h of treatment
（P＜0.01）；the percentage of cells in the early stage of apoptosis，the late stage of apoptosis and the early stage of necrosis，the
late stage of necrosis（except for the percentage of cells in the late stage of apoptosis and early stage of necrosis when treated with
YS-F 27 μg/mL for 72 h，the percentage of cells in the late stage of necrosis when treated with YS-F 3，9 μg/mL for 72 h were
decreased significantly）and the percentage of cells in S phase（except for YS-F 3 μg/mL for 72 h）and Sub-G0/G1 phase after
treated for 72 h were significantly higher than same group after 48 h of treatment，while the percentage of cells in G0/G1 and G2/M
phase were significantly lower than same group after 48 h of treatment（P＜0.01）. After treated with YS-F 9，27，81 μg/mL for 48
h，the reduction rate of cell mitochondrial membrane potential was increased significantly；YS-F 27，81 μ g/mL groups were
significantly higher than YS-F 9 μg/mL group，and YS-F 81 μg/mL group was significantly higher than YS-F 27 μg/mL group.
CONCLUSIONS：YS-F can inhibit the proliferation and promote the apoptosis of A549 cells by preventing cell transformation
from S phase to G2/M phase，and reducing mitochondrial membrane potential，in time-dependent or dose-dependent manner.
KEYWORDS Periplaneta americana；Non-small cell lung cancer cell；A549 cells；Proliferation；Apoptosis；Cell cycle；
Mitochondrial membrane potential

美洲大蠊（Periplaneta americana L.）属昆虫纲蜚蠊 CHEN PP等 研究发现，由美洲大蠊提取物制得的康复
[8]
[1]
科昆虫 ，最早记载于《神农本草经》，“主血瘀症坚，寒 新液可通过激活内质网应激和自噬来促进胃癌细胞的
热，破积聚，喉咽闭，内寒无子”。现代药理研究表明， 凋亡。本课题组参考上述文献，以美洲大蠊干燥虫体乙
[2]
美洲大蠊具有抗肿瘤、抑菌、修复受损组织等作用 [3-5] ；其 醇提取物为对象，经不同体积分数的甲醇梯度洗脱、冷
提取物对呼吸系统肿瘤细胞具有明显的抑制作用 。张 冻干燥后，制得各相应洗脱部位（以下简称“YS”）；在筛
[6]
丹等 报道，美洲大蠊多肽提取物可通过上调人肝癌细 选活性部位的基础上，考察其对人非小细胞肺癌 A549
[7]
胞 SMMC-7721 中促凋亡蛋白 Bax 的表达、下调抗凋亡 细胞增殖、细胞周期、线粒体凋亡等的影响，以期为美洲
蛋白Bcl-2的表达，从而发挥促进肝癌细胞凋亡的作用； 大蠊的进一步开发利用提供实验依据。

·402 · China Pharmacy 2020 Vol. 31 No. 4 中国药房 2020年第31卷第4期