目的 探讨两种丹参活性成分——隐丹参酮及丹酚酸 F 抑制肺癌增殖、迁移的作用及其潜在机制。 方法 体外培养小鼠肺癌 Lewis 细胞（LLC），采用 CCK-8 实验计算隐丹参酮及丹酚酸 F 抗肺癌的半数抑制浓度 （IC50）并以 IC50为依据，将 LLC 细胞分为对照组与实验组，对照组给予等体积溶剂，实验组分别给予不同浓度 隐丹参酮（5、10 、15 μmol·L-1 ）或丹酚酸 F（20、40、60 μmol·L-1 ）处理，干预 48 h。采用集落形成实验检测细 胞增殖能力；细胞划痕实验检测细胞迁移能力；乳酸、丙酮酸、ATP 含量测试盒检测有氧糖酵解产物生成情 况；Western Blot 实验检测有氧糖酵解酶的蛋白表达情况。为了进一步研究有氧糖酵解在丹参活性成分抗肺癌 中的作用，使用己糖激酶 II（HKII）的抑制剂 2-脱氧-D-葡萄糖（2-DG）及 3-溴丙酮酸（3-BP）进行干预，并构建 HKII 过表达或敲低的 LLC 细胞株，再次评价隐丹参酮及丹酚酸 F 抑制细胞增殖与迁移的作用。结果 隐丹参 酮抑制 LLC 细胞 24、48、72 h 的 IC50 分别为 29.38 、9.56、8.12 μmol·L-1 。丹酚酸 F 抑制 LLC 细胞 24、48、 72 h 的 IC50分别为 62.93 、42.12、31.99 μmol·L-1 。与对照组比较，隐丹参酮或丹酚酸 F 给药后，LLC 细胞集落 数量明显减少（P＜0.05，P＜0.01）；细胞迁移能力明显被抑制（P＜0.01）。同时，隐丹参酮与丹酚酸 F 给药后， LLC 细胞的有氧糖酵解产物乳酸、丙酮酸、ATP 含量均下降（P＜0.05，P＜0.01）；有氧糖酵解关键酶如 HKII、 血小板型磷酸果糖激酶（PFKP）、乳酸脱氢酶（LDHA）的蛋白表达水平均明显下调（P＜0.05，P＜0.01），其中 HKII 变化较明显。HKII 抑制剂 2-DG 及 3-BP 处理后，隐丹参酮与丹酚酸 F 抑制 LLC 细胞增殖和迁移的作用 明显增强（P＜0.05，P＜0.01）。降低 HKII 转录水平明显增强了隐丹参酮和丹酚酸 F 对 LLC 细胞增殖和迁移的 抑制效果（P＜0.01）；而过表达 HKII 则促进了 LLC 细胞的增殖和迁移，拮抗了隐丹参酮与丹酚酸 F 的抗肺癌 作用（P＜0.05，P＜0.01）。结论 丹参活性成分隐丹参酮及丹酚酸 F 可抑制 LLC 细胞的增殖与迁移，且此作用 与下调糖酵解关键酶 HKII 进而抑制肺癌细胞有氧糖酵解有关。

Objective To investigate the inhibitory effects of two active components from Salviae Miltiorrhizae Radix et Rhizoma—cryptotanshinone and salvianolic acid F—on lung cancer proliferation and migration， as well as their underlying mechanisms. Methods Mouse Lewis lung carcinoma （LLC） cells were cultured in vitro. The half-maximal inhibitory concentrations （IC50） of cryptotanshinone and salvianolic acid F against lung cancer were calculated using the CCK-8 assay. Based on the IC50 values，LLC cells were divided into control and experimental groups. The control group was treated with an equal volume of solvent，while the experimental groups were treated with different concentrations of cryptotanshinone （5，10，15 μmol·L-1 ） or salvianolic acid F （20，40，60 μmol·L-1 ） for 48 hours. Colony formation assay was used to assess cell proliferation ability；scratch wound healing assay was employed to evaluate cell migration； lactate， pyruvate， and ATP content assay kits were used to measure aerobic glycolysis product generation； and Western Blot was performed to detect the protein expression of key aerobic glycolysis enzymes. To further investigate the role of aerobic glycolysis in the anti-lung cancer effects of Salviae Miltiorrhizae Radix et Rhizoma active components， hexokinase II （HKII） inhibitors 2-deoxy-D-glucose （2-DG） and 3-bromopyruvate （3-BP） were applied，and HKIIoverexpressing or HKII-knockdown LLC cell lines were constructed to reassess the inhibitory effects of cryptotanshinone and salvianolic acid F on cell proliferation and migration. Results The IC50 values of cryptotanshinone against LLC cells at 24 hours， 48 hours， and 72 hours were 29.38， 9.56， and 8.12 μmol·L-1 ， respectively. The IC50 values of salvianolic acid F against LLC cells at 24 hours，48 hours，and 72 hours were 62.93，42.12，and 31.99 μmol·L-1 ， respectively. Compared with the control group，cryptotanshinone and salvianolic acid F treatment significantly reduced the number of LLC cell colonies （P＜0.05， P＜0.01） and inhibited cell migration （P＜0.01）. Additionally， cryptotanshinone and salvianolic acid F treatment decreased the levels of aerobic glycolysis products （lactate，pyruvate， and ATP）（P＜0.05，P＜0.01） and downregulated the protein expression of key aerobic glycolysis enzymes，including HKII，platelet-type phosphofructokinase （PFKP），and lactate dehydrogenase （LDHA）（P＜0.05，P＜0.01），with the most significant change observed in HKII. After treatment with HKII inhibitors 2-DG and 3-BP， the inhibitory effects of cryptotanshinone and salvianolic acid F on LLC cell proliferation and migration were significantly enhanced （P＜0.05， P＜0.01）. Reducing HKII transcriptional levels markedly strengthened the inhibitory effects of cryptotanshinone and salvianolic acid F on LLC cell proliferation and migration （P＜0.01）， whereas HKII overexpression promoted LLC cell proliferation and migration， counteracting the anti-lung cancer effects of cryptotanshinone and salvianolic acid F （P＜0.05， P＜0.01）. Conclusion The active components cryptotanshinone and salvianolic acid F from Salviae Miltiorrhizae Radix et Rhizoma inhibit LLC cell proliferation and migration，and this effect is associated with the downregulation of the key glycolysis enzyme HKII，thereby suppressing aerobic glycolysis in lung cancer cells.

R285.5

国家自然科学基金项目（81903943）；广东省自然科学基金项目（2023A1515011343）。