Mechanism Study of Xiaoyan Decoction in the Treatment of Non-small Cell Lung Cancer Through Glycometabolic Reprogramming

Abstract [Objective] To investigate the effects of Xiaoyan Decoction ( XYT) on autophagy and glucose metabolism reprogramming in Non-small Cell Lung Cancer(NSCLC) cells and to explore its mechanism of action. [Methods] ① Screen the half maximal inhibitory concentration (IC50) of XYT-containing serum using Cell Counting Kit 8 (CCK8); ② After pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) and the activator Rapamycin (RAPA) and intervention with XYT, transmission electron microscopy was used to observe the overall autophagy status of A549 cells, and Western Blot was used to detect changes in Sequestosome1(P62) and Microtubule-associated protein 1 light chain 3(LC3); ③ The kit was used to detect glucose uptake and lactate production, Seahorse was used to assess cellular energy metabolism, and Western Blot was used to assess the expression of Hypoxia inducible factor-1α(HIF-1α) protein in A549 cells after XYT drug-containing serum intervention alone and in combination with RAPA intervention. [Results] ①XYT-containing serum can slow down cell proliferation in a concentration-dependent manner, with a 30% concentration of XYT-containing serum for 24 hours being the optimal concentration and duration of intervention. ②Following XYT intervention, the number of autophagosomes—a characteristic double-membrane structure of autophagy—significantly increased in A549 cells. Western blot analysis revealed a significant downregulation of P62 expression and a significant upregulation of LC3-II/LC3-I protein expression. ③After XYT intervention, glucose uptake and lactate production in A549 cells were significantly reduced, glycolytic rate was significantly slowed, and maximum glycolytic capacity was also significantly reduced. Western blot analysis showed a significant downregulation of HIF-1α expression. Additionally, Oxygen consumption rate(OCR) assay results indicated that O₂ consumption significantly increased after XYT intervention, with parameters such as basal respiration and maximal respiration significantly up-regulated. These effects were further enhanced by co-treatment with autophagy modulators 3-MA/RAPA. [Conclusion] XYT can inhibit A549 cell proliferation, upregulate cellular autophagy levels, regulate HIF-1α expression, shift the metabolic phenotype from glycolysis to aerobic oxidation, reprogram glucose metabolism, and thereby exert an inhibitory effect on cell proliferation.