基于网络药理学和细胞实验探讨小檗碱治疗肺癌的作用机制

来源:用户上传      作者:邹宇翔 柳鑫 李为

　　[摘要]目的探究小檗碱对人非小细胞肺腺癌细胞A549的增殖抑制和凋亡作用机制。方法采用在线数据库获取小檗碱和肺癌的交集靶点，构建蛋白互作（PPI）网络分析，对靶点进行基因本体功能（GO）及京都基因和基因组百科全书（KEGG）通路富集分析。以人非小细胞肺腺癌细胞A549为受试对象开展实验验证，采用细胞计数试剂法（CCK-8）筛选小檗碱对细胞增殖抑制作用时间和浓度;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法（TUNEL）检测细胞凋亡;蛋白质印迹法（Western blotting）检测p53、Bax和Bcl-2蛋白表达水平。结果网络药理学分析得到交集靶点92个，富集到的信号通路主要包括p53和凋亡（Apotosis）等信号通路。实验验证发现，不同浓度的小檗碱均能不同程度地抑制肺腺癌A549细胞的增殖（P<0.01）;干预24 h时，IC50值为8.51 μg/ ml，且小檗碱能显著上{p53和Bax蛋白，下调Bcl-2蛋白的表达水平（P<0.01）。结论小檗碱可以作为一种潜在的抗肿瘤药物进一步研究。
　　[关键词]小檗碱;肺癌;网络药理学;细胞凋亡
　　[中图分类号]R285[文献标识码]A[文章编号]2095-0616（2022）13-0086-05
　　To explore the mechanism of berberine in the treatment of lung cancer based on network pharmacology and cell experiment
　　ZOU Yuxiang1LIU Xin2LI Wei1
　　l.Department of Pharmacy，Tongji Hospital，Tongji Medical College，Huazhong University of Science and Technology，Hubei，Wuhan 430030，China;2. Department of Pharmacy，Wuhan Fourth Hospital，Hubei，Wuhan 430033，China
　　[Abstract] Objective To explore the mechanism of berberine on the proliferation inhibition and apoptosis of huaman non-small cell lung adenocarcinoma CELL A549. Methods The intersection targets of berberine and lung cancer were obtained from online database，protein interaction （PPI）network analysis was constructed，and gene body function （GO）and Kyoto Encyclopedia of Genes and Genomes （KEGG）pathway enrichment analysis were performed on the targets. Cell counting reagent （CCK-8）was used to screen the time and concentration of berberine inhibiting cell proliferation in huaman non-small cell lung adenocarcinoma CELL A549. Deoxyribonucleotide terminal transferase mediated tdt-mediated dutp nick end labeling （TUNEL）was used to detect cell apoptosis. The expression levels of p53，Bax and Bcl-2 proteins were detected by Western blotting. Results The network pharmacology analysis showed 92 intersecting targets，and the enriched signaling pathways mainly included p53 and Apoptosis signaling pathways. The experimental verification found that different concentrations of berberine can inhibit the proliferation of A549 cells to different degrees （P<0.01），and the IC50 value was 8.51 μg/ml at 24 h;Western blotting results show that berberine can up-regulate the expression levels of p53 and Bax proteins，and down-regulate Bcl-2 proteins （P<0.01）. Conclusion Berberine can can be used as a potential anti-tumor drug for further research.
　　[Key words] Berberine;Lung cancer;Network pharmacology;Apoptosis
　　肺癌是世界重大公共卫生问题，其发病率与病死率均占恶性肿瘤的首位，全球范围内的肺癌中有80%～85%被归为非小细胞肺癌，临床治疗主要采用手术、化学疗法和放射疗法等[1]。化学药物和分子靶向药物的临床应用大大延长了晚期肺癌患者的生存期并改善其生活质量[2]，然而长期使用会产生耐药性问题，因此，寻找新的治疗肺癌有效药物成为科学研究的重要方向[3-4]。小檗碱是中药黄连发挥临床疗效的重要活性成分。研究发现，小檗碱具有抗肿瘤、抗菌和调节血糖等作用[5-8]，然而其治疗肺癌的机制尚不清楚。本研究采用网络药理学探究其治疗肺癌的潜在作用机制并进行初步的实验验证，以期为临床使用提供依据。

nlc202208181137

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