Fig 2: Isorhamnetin induces GBC cell apoptosis. (A) The stained NOZ and GBC-SD cells exposed to isorhamnetin (0, 40, 60, and 80 μM). (B) Apoptotic cell proportions are expressed as the mean ± SD. (C) The apoptotic nuclear morphology alterations were assessed using Hoechst 33342 staining via a fluorescence microscope. (D) The expression and (E) statistic results of cleaved PARP, PARP, BAX, cleaved caspases 9, caspases 9, BCL-2, cleaved caspases 3, and caspases 3 in NOZ and GBC-SD after isorhamnetin treatment. *p < 0.05, **p < 0.01 relative to the control group.

Fig 3: Isorhamnetin inhibits GBC cell proliferation and metastasis via the PI3K/AKT pathway. After pretreatment with SC79 for 1 h, the expression levels of (A) MMP-2, Slug, MMP-9, E-cadherin, N-cadherin, TWIST1, (B) cleaved PARP, PARP, BAX, cleaved caspases 9, caspases 9, BCL-2, cleaved caspases 3, caspases 3, (C) cyclin B1, CDK1, p27, p53, p-PI3K, p-AKT1, PIK3CA, and AKT were detected and shown with statistic results. *p < 0.05, **p < 0.01, ***p < 0.001 relative to the control group.

Fig 4: miR-93-5p inhibited autophagy and apoptosis of IL-1β-treated chondrocytes by targeting ADAMTS9 to activate the PI3K/AKT/mTOR pathway. Note: (a) MDC staining was used to detect the autophagy of control, IL-1β, IL-1β + miR-93-5p inhibitor, IL-1β + si-ADAMTS9, and IL-1β + miR-93-5p inhibitor + si-ADAMTS9 group in chondrocytes. (b) Flow cytometry was used to detect the apoptosis of control, IL-1β, IL-1β + miR-93-5p inhibitor, IL-1β + si-ADAMTS9, and IL-1β + miR-93-5p inhibitor + si-ADAMTS9 group in chondrocytes. (c) Western blot assay was used to detect the expression of PI3K, P-mTOR, P-AKT, Bcl-2/Bax, and LC3B-I/II expression of control, IL-1β, IL-1β + miR-93-5p inhibitor, IL-1β + si-ADAMTS9, and IL-1β + miR-93-5p inhibitor + si-ADAMTS9 group.