Xu Yan-Wei, Geng Sheng-Nan, Wang Yue-Hua, DU Gang-Jun
Zhengzhou University of Industrial Technology Xinzheng 451150,China Luoyang Third People's Hospital Luoyang 471000,China.
Zhengzhou University of Industrial Technology Xinzheng 451150,China.
Zhongguo Zhong Yao Za Zhi. 2021 Mar;46(6):1480-1489. doi: 10.19540/j.cnki.cjcmm.20201027.402.
This study is to observe whether platycodin D has the guiding role in treatment of mouse lung cancer with doxorubicin and explore its guiding mechanism. In vitro, platycodin D and doxorubicin(alone or in combination) were added into Lewis lung cancer(LLC) cells to detect the cell proliferation and doxorubicin uptake. Cell morphological changes were analyzed by cell holographic analysis system; cell gap junctional intercellular communication(GJIC) was tested by fluorescent yellow tracer; lyso-tracker red was used to examine lysosomal function; LC-3 B(Light chain 3 beta)and P62(heat shock 90-like protein)staining were used to test auto-phagy and autophagic degradation respectively; and P-glycoprotein(P-gp) expression was examined by Western blot. In vivo, lung solid tumor was formed in mouse LLC cells via intravenous injection. Platycodin D and doxorubicin(alone or in combination) were used to treat tumor-bearing mice for four weeks, and then the tumor size was examined, mouse survival time was recorded, doxorubicin uptake in lung tissues was tested, and lung tissues were stained for observation by HE(hematoxylin-eosin) and immunohistochemistry. The results showed that platycodin D at the experimental concentration had no effect on LLC cell proliferation but decreased LLC cell volume, promoted the cells to uptake doxorubicin and enhanced the inhibitory action of doxorubicin on cell proliferation. Platycodin D could promote GJIC and lysosomal function, increase autophagy and autophagic degradation and suppress P-gp expression. Platycodin D at the experimental dose in this study had no effect on LLC lung solid tumors in mice, increased doxorubicin uptake in lung tissues and enhanced the therapeutic efficacy of doxorubicin on lung solid tumors. Platycodin D could improve the extracellular matrix deposition in lung solid tumors, decreased the lung mucin 5 AC secretion and pulmonary vessel permeability. In summary, platycodin D had the guiding role in treating mouse lung cancer with doxorubicin, and its guiding mechanism may be associated with the promotion of cell communication, lysosomal function, and improvement of extracellular environment.
本研究旨在观察桔梗皂苷D对阿霉素治疗小鼠肺癌是否具有导向作用，并探讨其导向机制。体外实验中，将桔梗皂苷D和阿霉素（单独或联合使用）加入Lewis肺癌（LLC）细胞中，检测细胞增殖和阿霉素摄取情况。通过细胞全息分析系统分析细胞形态变化；用荧光黄示踪剂检测细胞间隙连接细胞间通讯（GJIC）；用溶酶体示踪红检测溶酶体功能；用LC-3 B（轻链3β）和P62（热休克90样蛋白）染色分别检测自噬和自噬降解；用蛋白质免疫印迹法检测P-糖蛋白（P-gp）表达。体内实验中，通过静脉注射在小鼠LLC细胞中形成肺实体瘤。用桔梗皂苷D和阿霉素（单独或联合使用）治疗荷瘤小鼠4周，然后检查肿瘤大小，记录小鼠存活时间，检测肺组织中阿霉素摄取情况，并用苏木精-伊红（HE）染色和免疫组织化学染色观察肺组织。结果显示，实验浓度的桔梗皂苷D对LLC细胞增殖无影响，但可减小LLC细胞体积，促进细胞摄取阿霉素，并增强阿霉素对细胞增殖的抑制作用。桔梗皂苷D可促进GJIC和溶酶体功能，增加自噬和自噬降解，并抑制P-gp表达。本研究中实验剂量的桔梗皂苷D对小鼠LLC肺实体瘤无影响，但可增加肺组织中阿霉素摄取，增强阿霉素对肺实体瘤的治疗效果。桔梗皂苷D可改善肺实体瘤细胞外基质沉积，减少肺黏蛋白5 AC分泌和肺血管通透性。综上所述，桔梗皂苷D对阿霉素治疗小鼠肺癌具有导向作用，其导向机制可能与促进细胞通讯、溶酶体功能及改善细胞外环境有关。
