Frontiers in pharmacology · 2026
Combination of polygonatum sibiricum and hedyotis diffusa induces pyroptosis in non-small cell lung cancer cells via the TLR4/NLRP3/Caspase-1/GSDMD pathway: an in vitro and in vivo study using medicated serum and herbal extract.
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Mao Lei, Jiang Lingying, Zhou Ye, Xu Shouhang, Lou Tingting, Lu Weiyong, Zhu Gaoyuan, Chen Jianwu
Аннотация
OBJECTIVE: To study the effects of Polygonatum-Hedyotis diffusa on non-small cell lung cancer and the mechanism of action.
METHODS: The CCK-8 assay was used to investigate the effects of different ratios of Polygonatum-Hedyotis diffusa containing serum on the proliferation of A549 cells, and the optimal ratios were screened out for subsequent experiments. The scratch and invasion assays were used to evaluate the inhibition of migration and invasion of A549 cells by Polygonatum-Hedyotis diffusa containing serum. Flow cytometry was used to analyze the effect of Polygonatum-Hedyotis diffusa containing serum on the apoptosis of A549 cells, and Western blot assay was used to analyze the protein expression levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18. Finally, mice were injected subcutaneously with A549 cells in vivo to form solid tumors to further validate the in vivo inhibitory effect of Polygonatum-Hedyotis diffusa on non-small-cell lung cancer, and Western blot experiments were used to re-validate the levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 protein expression in tumor cells.
RESULT: CCK-8 experiments were used to demonstrate that the 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum had the best inhibitory effect on A549 cells. Scratch and invasion experiments verified that the 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum inhibited the migration and invasion ability of A549. Flow cytometry results illustrated that 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum induced apoptosis in A549 cells, and Western blot experiments verified that the expression levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 proteins were up-regulated. In vivo animal experiments further validated Polygonatum-Hedyotis diffusa for the treatment of non-small cell lung cancer, and Western blot verified the upregulation of the protein expression levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 in tumor cells.
CONCLUSION: 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum can inhibit the proliferation of A549 cells through cellular pyroptosis, providing a new idea for the treatment of non-small cell lung cancer. The upregulation of these proteins suggests the involvement of the pathway, but causal evidence is not provided in this study.
METHODS: The CCK-8 assay was used to investigate the effects of different ratios of Polygonatum-Hedyotis diffusa containing serum on the proliferation of A549 cells, and the optimal ratios were screened out for subsequent experiments. The scratch and invasion assays were used to evaluate the inhibition of migration and invasion of A549 cells by Polygonatum-Hedyotis diffusa containing serum. Flow cytometry was used to analyze the effect of Polygonatum-Hedyotis diffusa containing serum on the apoptosis of A549 cells, and Western blot assay was used to analyze the protein expression levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18. Finally, mice were injected subcutaneously with A549 cells in vivo to form solid tumors to further validate the in vivo inhibitory effect of Polygonatum-Hedyotis diffusa on non-small-cell lung cancer, and Western blot experiments were used to re-validate the levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 protein expression in tumor cells.
RESULT: CCK-8 experiments were used to demonstrate that the 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum had the best inhibitory effect on A549 cells. Scratch and invasion experiments verified that the 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum inhibited the migration and invasion ability of A549. Flow cytometry results illustrated that 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum induced apoptosis in A549 cells, and Western blot experiments verified that the expression levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 proteins were up-regulated. In vivo animal experiments further validated Polygonatum-Hedyotis diffusa for the treatment of non-small cell lung cancer, and Western blot verified the upregulation of the protein expression levels of TLR4, NLRP3, Caspase-1, GSDMD, IL-1β, and IL-18 in tumor cells.
CONCLUSION: 3:1 ratio of Polygonatum-Hedyotis diffusa containing serum can inhibit the proliferation of A549 cells through cellular pyroptosis, providing a new idea for the treatment of non-small cell lung cancer. The upregulation of these proteins suggests the involvement of the pathway, but causal evidence is not provided in this study.