目的 研究忍冬藤Lonicera japonica提取物对骨肉瘤细胞增殖和凋亡的作用及机制。方法 采用超高效液相色谱-飞行时间质谱联用技术（UPLC-Q-TOF/MS）表征忍冬藤提取物的化学成分；采用MTT法检测忍冬藤提取物对人骨肉瘤细胞HOS、143B以及人肾皮质近曲小管上皮细胞HK-2存活率的影响；采用荧光显微镜观察忍冬藤提取物对HOS、143B细胞凋亡的影响；采用Annexin V/PI双染法检测忍冬藤提取物对HOS细胞凋亡的影响；采用JC-1染色法检测忍冬藤提取物对HOS细胞线粒体膜电位的影响；采用Western blotting法检测忍冬藤提取物对HOS细胞B细胞淋巴瘤2相关X蛋白（bcl-2 associated X，Bax）、B细胞淋巴瘤2（B cell lymphoma-2，Bcl-2）、活化的半胱氨酸蛋白酶-3（cleaved Caspase-3）、活化的多聚腺苷二磷酸核糖聚合酶（cleaved poly ADP-ribose polymerases，cleaved PARP）、cleaved Caspase-9和细胞色素C（cytochrome-c，Cyt-C）蛋白表达的影响。结果 忍冬藤提取物中鉴定出有机酸类、三萜皂苷类、黄酮类、环烯醚萜类和氨基酸类5种类型的36种化学成分。忍冬藤提取物抑制HOS和143B细胞存活率，显著诱导HOS细胞凋亡（P<0.01），Caspase-3抑制剂Z-VAD-FMK显著抑制忍冬藤提取物诱导的HOS细胞凋亡（P<0.05、0.01）；忍冬藤提取物显著提高HOS细胞Caspase-3活性（P<0.01），显著改变HOS细胞线粒体膜电位（P<0.05、0.01），显著上调HOS细胞Bax、cleaved Caspase-3、cleaved PARP、cleaved Caspase-9、Cyt-C和Bax/Bcl-2蛋白表达水平（P<0.05、0.01），显著下调Bcl-2蛋白表达水平（P<0.05、0.01）。结论 忍冬藤提取物能够抑制骨肉瘤细胞增殖，并通过线粒体凋亡途径诱导细胞凋亡，其机制可能与下调Bcl-2蛋白表达，上调Bax、cleaved Caspase-3、cleaved PARP、cleaved Caspase-9和Cyt-C蛋白表达，激活Caspase级联反应有关。

Objective To investigate the proliferation and apoptosis effects and mechanism of Lonicera japonica extract on human osteosarcoma cells. Methods Chemical constituents of L. japonica extract were characterized by UPLC-Q-TOF/MS. MTT method was used to detect the effect of L. japonica extract on survival rate of human osteosarcoma cells HOS and 143B and human renal cortex proximal tubule epithelial cells HK-2; Fluorescence microscope was used to observe the effect of L. japonica extract on apoptosis of HOS and 143B cells; Annexin V/PI double staining method was used to detect the effect of L. japonica extract on apoptosis of HOS cells; JC-1 staining method was used to detect the effect of L. japonica extract on mitochondrial membrane potential of HOS cells; Western blotting method was used to detect the effect of L. japonica extract on expressions of B cell lymphoma 2 associated X protein (Bax), B cell lymphoma 2 (b-cell lymphoma-2, Bcl-2), activated cysteine protease-3 (cleaved Caspase-3), cleaved poly ADP-ribose polymerase (cleaved PARP), cleaved Caspase-9 and cytochrome-c (Cyt-C) in HOS cells. Results A total of 36 chemical components of five types, including organic acids, triterpene saponins, flavonoids, iridoids and amino acids, were identified in L. japonica extract. L. japonica extract inhibited the survival rate of HOS and 143B cells, and significantly induced HOS cells apoptosis (P < 0.01). Caspase-3 inhibitor Z-VAD-FMK significantly inhibited HOS cells apoptosis induced by L. japonica extract (P < 0.05, 0.01); L. japonica extract significantly increased Caspase-3 activity in HOS cells (P < 0.01), changed mitochondrial membrane potential of HOS cells (P < 0.05, 0.01), increased expressions of Bax, cleaved Caspase-3, cleaved PARP, cleaved Caspase-9, Cyt-C and Bax/Bcl-2 in HOS cells (P < 0.05, 0.01), down-regulated Bcl-2 expression (P < 0.05, 0.01). Conclusion L. japonica extract can inhibit the proliferation of osteosarcoma cells and induce apoptosis through mitochondrial apoptosis. The mechanism may be related to down-regulation of Bcl-2 expression, up-regulation of Bax, cleaved Caspase-3, cleaved PARP, cleaved Caspase-9 and Cyt-C expression, and activation of Caspase cascade reaction.

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国家中医管理局名医验方评价与转化重点研究室开放课题（NZYJDMF-2020001）；江苏省自然科学基金资助项目（BK20191498）；江苏省卫生健康委医学科研项目重点项目（ZDB2020020）；浦口区科技发展社会事业项目（S2020-14）；江苏卫生健康职业学院校级课题重点项目（JKA201916）