中药材2023，Vol.46Issue(7)：1760-1765,6.DOI:10.13863/j.issn1001-4454.2023.07.031
岩黄连总生物碱抑制M2型巨噬细胞抗小鼠肝癌实验研究
Experimental Study of Corydalis saxicola Total Alkaloids Inhibiting M2-Type Macrophages Against Mice Liver Cancer
摘要
Abstract
Objective:To investigate the inhibitory effect of total alkaloids of Corydalis saxicola on M2-type macrophages against solid tumor growth of mice H22 hepatoma cells by negatively regulating PI3K/AKT/mTOR signaling pathway.Methods:Mouse H22 hepa-toma cell solid tumor model was established.After giving high,medium and low doses(100,50,25 mg/kg)of total alkaloids of Coryda-lis saxicola for 10 days,tumor,spleep and thymus weight was measured,organ coefficient was calculated,hematoxylin-eosin(HE)stai-ning was performed to observe tumor histomathological changes.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect mRNA expression of CD206,a marker of M2-type macrophage in tumor tissue.RAW264.7 cells were cul-tured in vitro,induced M2 polarization by IL-4 and IL-13(20 ng/mL),and treated with high,medium and low concentrations of Coryda-lis saxicola total alkaloids(25,12.5,6.25 μg/mL)for 24 hours.CD206 expression of M2-type macrophage marker was detected by flow cytometry.The mRNA expressions of CD206,Arg-1 and IL-10 were detected by RT-qPCR.The protein expressions of PI3Kp110,p-AKT and p-mTOR in RAW264.7 cells were detected by Western Blot.Results:Animal experiments showed that high and medium doses of total alkaloids of Corydalis saxicola inhibited the growth of H22 hepatocellular carcinoma solid tumor and significantly increased spleen coefficient and thymus coefficient(P<0.05),in HE stained sections,the number of tumor cell infiltration decreased,CD206 mRNA ex-pression in necrotic tumor tissues was significantly decreased(P<0.05 or P<0.01).Cell experiment results showed that total alkaloids in high and medium concentration groups could significantly reduce the positive expression of CD206 in M2-type macrophage markers,the mRNA expressions of CD206,Arg-1 and IL-10 and the protein expression of PI3Kp110,p-AKT and p-mTOR in PI3K/AKT/mTOR signaling pathway(P<0.05 or P<0.01).Conclusion:The total alkaloids of Corydalis saxicola can inhibit the M2-type polarization of macrophages by negatively regulating PI3K/AKT/mTOR signaling pathway,and play an anti-growth role in the growth of mouse H22 liv-er cancer cell solid tumor.关键词
岩黄连总生物碱/肝癌/M2型巨噬细胞/PI3K/AKT/mTOR信号通路Key words
Total alkaloids of Corydalis saxicola Bunting/Hepatocellular carcinoma/M2 Macrophages/PI3K/AKT/mTOR signal-ling pathway分类
医药卫生引用本文复制引用
苏倩,陈好然,陆洁,黄丽贞,梁健钦,张文涛..岩黄连总生物碱抑制M2型巨噬细胞抗小鼠肝癌实验研究[J].中药材,2023,46(7):1760-1765,6.基金项目
广西高校中青年教师科研基础能力提升项目(2022KY0306) （2022KY0306）
国家自然科学基金资助项目(81960872) （81960872）
广西高校中药药理重点实验室资助项目(05J15185) （05J15185）
