| 碱制法炮制可显著增强斑蝥的抗肿瘤作用 |
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| 引用本文： | 李 娴,李姗姗,庞金龙,黄富豪,郭 滨,刘 浩.碱制法炮制可显著增强斑蝥的抗肿瘤作用[J].南方医科大学学报,2020,40(9):1332-1339. |
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| 作者姓名： | 李 娴 李姗姗 庞金龙 黄富豪 郭 滨 刘 浩 |
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| 摘 要： | 目的 研究碱制法炮制前后斑蝥对人肺癌A549细胞增殖、迁移、侵袭、凋亡的影响。方法 提取生品斑蝥中的斑蝥素和碱制斑蝥中的斑蝥酸钠。以人非小细胞肺癌A549细胞为研究对象，生斑蝥与碱制斑蝥处理细胞后，采用CCK-8法检测药物对细胞增殖活性的影响，划痕实验观察细胞迁移作用，Transwell实验观察细胞侵袭作用，WB观察药物对A549细胞MMP1、MMP2蛋白表达的影响，ELISA法检测细胞炎症因子IFN-γ、IL-1β和TNF-α水平，AnnexinV/PI染色观察细胞凋亡情况。结果 生斑蝥与碱制斑蝥均可使A549细胞活力明显降低；生斑蝥与碱制斑蝥均能抑制A549细胞迁移，且生斑蝥高浓度组划痕愈合程度最低，与生斑蝥高浓度组相比，差异具有显著性（P<0.01）；碱制斑蝥较生斑蝥抑制A549细胞侵袭作用更明显，差异具有显著性（P<0.01）；且生斑蝥与碱制斑蝥均可抑制 MMP1和MMP2蛋白的表达，显著上调炎症因子IFN-γ水平，但对IL-1β、TNF-α含量则没
有显著影响，Annexin V/PI双染荧光拍照发现生斑蝥组与碱制斑蝥组红绿荧光强度均增加，且碱制斑蝥组荧光增加更明显。结论 碱制法炮制后斑蝥的抗癌作用显著提高，具有抑制A549细胞增殖，侵袭和迁移的作用，其机制可能与下调MMP1和MMP2的表达，促进凋亡及上调炎症因子IFN-γ水平有关。
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| 关 键 词： | 碱制斑蝥 凋亡 炎症因子 迁移 侵袭 |
Alkaline processing of cantharidin can significanty improve the antitumor activity of
cantharidin |
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| Abstract: | Objective To assess the changes in the effects of cantharides after alkaline processing on proliferation, migration,
invasion, and apoptosis of human lung cancer A549 cells. Methods Human non-small cell lung cancer A549 cells were treated
with cantharis extract (CTE) from raw cantharides and alkali processed cantharis extract (ACE). The proliferation of the cells
was detected with CCK-8 assay, and the cell migration and invasion were assessed using wound healing assay and Transwell
assay, respectively. The expressions of MMP1 and MMP2 in the cells were detected using Western blotting, the contents of IFN-γ,
IL-1β and TNF-α were measured with ELISA, and cell apoptosis was analyzed with annexinV/PI fluorescent staining. Results
Both CTE and ACE significantly reduced the viability and inhibited the migration of A549 cells, and high-dose ACE produced
a significantly stronger inhibitory effect on cell migration than high-dose CTE (P<0.01). ACE showed more potent inhibitory
effect than CTE on the invasion of A549 cells (P<0.01). Both CTE and ACE inhibited the expressions of MMP1 and MMP2 and
up-regulated the level of IFN-γ without significantly affecting the levels of IL-1β and TNF-α. Annexin V/PI staining showed
that both CTE and ACE caused apoptosis of A549 cells, but ACE had a stronger proapoptotic effect. Conclusion Processing
with sodium hydroxide can significantly improve the antitumor activity of cantharides, which inhibits the proliferation,
migration and invasion of A549 cells possibly by down-regulating the expressions of MMP1 and MMP2, promoting apoptosis
and increasing the level of IFN-γ. |
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| Keywords: | alkaline processed cantharis apoptosis inflammatory factors migration invasion |
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