目的 考察人参皂苷Rh₂对人非小细胞肺癌A549及H460细胞增殖的影响，并进一步探讨其作用机制。方法 人参皂苷Rh₂处理A549及H460细胞，采用CCK-8法及克隆形成实验检测药物对细胞增殖的作用；观察药物对细胞形态的影响，利用Annexin V-FITC/碘化丙啶（PI）双染法检测细胞凋亡情况；利用乳酸检测试剂盒检测药物对细胞糖酵解产物即乳酸生成的影响；采用Western blotting法检测药物对线粒体凋亡蛋白[B淋巴细胞瘤2（B-cell lymphoma 2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、半胱氨酸蛋白酶-3（Caspase-3）]，糖酵解调节通路信号传导与转录激活因子3（signal transducer and activator of transcription 3，STAT3）/c-myc及糖酵解途径关键酶丙酮酸激酶M2（pyruvate kinase M2，PKM2）、乳酸脱氢酶A（lactate dehydrogenase A，LDHA）表达的影响。上调c-myc表达后，观察人参皂苷Rh₂对A549及H460细胞乳酸水平的影响。结果 人参皂苷Rh₂可明显抑制A549及H460细胞的增殖能力及克隆形成能力（P<0.05、0.01、0.001），呈剂量和时间相关性。人参皂苷Rh₂明显改变A549及H460细胞形态并诱导细胞凋亡（P<0.05），显著降低乳酸生成量（P<0.05、0.01）。人参皂苷Rh₂可显著下调Bcl-2/Bax、PKM2、LDHA蛋白表达水平（P<0.05、0.01），上调Caspase-3蛋白表达水平（P<0.05），并抑制STAT3/c-myc通路活性（P<0.05、0.01）。上调c-myc表达后，人参皂苷Rh₂对A549及H460细胞乳酸水平的抑制作用减弱（P<0.05、0.01）。结论 人参皂苷Rh₂显著抑制A549及H460细胞的增殖，其机制可能是通过调控线粒体凋亡蛋白、抑制STAT3/c-myc通路及糖酵解关键酶的表达干扰糖酵解，最终导致细胞凋亡。

Objective To study the effect and mechanism of ginsenoside Rh₂ on proliferation of human non-small cell lung cancer A549 and H460 cells. Methods A549 and H460 cells were treated with ginsenoside Rh₂, CCK-8 assay and colony formation experiment were used to detect the effect of ginsenoside Rh₂ on cell proliferation; Effect of ginsenoside Rh₂ on cell morphology was observed; Cell apoptosis were detected by Annexin V-FITC/PI dye method; Level of glycolytic products (lactic acid) was tested by lactic acid detection kit. Protein expression levels of mitochondrial apoptotic protein B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), Caspase-3 and glycolysis regulation pathway signal transducer and activator of transcription 3 (STAT3)/c-myc and glycolysis key enzymes pyruvate kinase M2 (PKM2) and lactate dehydrogenase A (LDHA) were detected by Western blotting. Effect of ginsenoside Rh₂ on lactate level in A549 and H460 cells after up-regulating c-myc expression were observed. Results Ginsenoside Rh₂ significantly inhibited the proliferation and colony formation ability of A549 and H460 cells in a concentration and time-dependent way (P < 0.05, 0.01, 0.001). Ginsenoside Rh₂ altered cell morphology and significantly induced cell apoptosis (P < 0.05) and reduced the levels of lactic acid (P < 0.05, 0.01). Ginsenoside Rh₂ down-regulated Bcl-2/Bax, PKM2 and LDHA protein expressions, up-regulated Caspase-3 expression (P < 0.05) and inhibited STAT3/c-myc pathway activity (P < 0.05, 0.01). The inhibitory effect of ginsenoside Rh₂ on lactate levels in A549 and H460 cells was abolished after up-regulating c-myc expression. Conclusion Ginsenoside Rh₂ has a significant inhibitory effect on proliferation of A549 and H460 cells. The mechanism may be through regulating mitochondrial apoptotic proteins, inhibiting STAT3/c-myc pathway and reducing glycolysis key enzyme expression to interfere glycolysis, and then leading to cells apoptosis.

R285.5

国家自然科学基金资助项目（82103343）；吉林省中医药科技项目（2021095）；吉林省卫生计生委科研项目（2018J021）；吉林省科技厅项目（202002063JC）