Effect of acacetin on inhibition of apoptosis in Helicobacter pylori-infected gastric epithelial cell line

Figure 1 Significant morphological alterations in GES-1 cells following exposure to Helicobacter pylori for 24 h and 48 h. A: 24 h; B: 48 h. × 40 magnification.

Figure 2 Helicobacter pylori infection reduces the viability of GES-1 gastric epithelial cells. A and B: Stimulation of cells for 24 h or 48 h at specified ratios of cells to Helicobacter pylori (H. pylori); C: Viability of GES-1 cells treated with different ratios of H. pylori over various time periods. The viability of the control group (GES-1 cells not stimulated with H. pylori) was set at 100%. Significant differences were observed: ^aP < 0.05, ^bP < 0.01, ^cP < 0.001, ^dP < 0.0001 vs control group.

Figure 3 Acacetin inhibits the decline in cell viability and lactate dehydrogenase release induced by Helicobacter pylori infection. A: Cells were pre-treated with specified concentrations of acacetin extract for 2 h, followed by stimulation with Helicobacter pylori (H. pylori) [multiplicity of infection (MOI) = 1:100] for 24 h; B: After pretreatment with specified concentrations of acacetin extract for 2 h and subsequent stimulation with H. pylori (MOI = 1:100) for 24 h, the amount of lactate dehydrogenase released into the cell culture supernatant was measured. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001, ^dP < 0.0001 vs control group; ^eP < 0.05, ^fP < 0.01, ^gP < 0.001, ^hP < 0.0001 vs H. pylori group.

Figure 4 Migration analysis was performed through a wound healing assay. A: Images of the wound region captured at four distinct time intervals (0, 12, 24, and 48 h). × 40 magnification; B: The area of wound closure was analyzed and converted into migration rates using ImageJ software. HP: Helicobacter pylori.

Figure 5 Acacetin extract inhibits Helicobacter pylori-induced apoptosis in GES-1 cells. A and B: Protein levels of cleaved caspase-3, Bcl-2, and Bax were analyzed by immunoblotting following pre-treatment with acacia extract (10 µmol/L, 20 µmol/L) and subsequent stimulation with Helicobacter pylori(H. pylori) (cell to H. pylori ratio of 1:100) for 24 h. GAPDH served as an internal control, with the ratio in the blank control group set to 1; C and D: Apoptosis levels in the blank group, H. pylori-infected group, and H. pylori plus acacetin extract group were measured by flow cytometry; E: Representative images of cells stained with TUNEL and DAPI. × 40 magnification; F: Percentage of apoptotic cells quantified as the apoptosis index.