Effect of Zuojin Formula on proliferation and apoptosis of human GES-1 cells infected by Helicobacter pylori / 中草药

ABSTRACT

Objective: To study the effect of Zuojin Formula on proliferation and apoptosis of human normal gastric epithelial cells (GES-1) cells infected by Helicobacter pylori. Methods: GES-1 cells were infected by H. pylori at different multiplicity of infection (1∶1, 50∶1, 100∶1, 200∶1, 300∶1) for 12, 24, 48 h; GES-1 cells infected by H. pylori were treated with different concentrations (0.5, 1.0, 2.0, 4.0 μg/mL) of Zuojin Formula, and the cells were harvested after 12, 24, and 48 h. The proliferation activity of cells was detected by CCK-8, and the apoptosis rate of GES-1 cells was measured by Anexin V-FITC apoptosis detecting kit and flow cytometry. Moreover, the expression of Caspase-3 was detected by Western blot and the morphological changes of apoptotic cells were detected by Hochest staining. Results: After infected by H. pylori, the cell viability showed a descending trend while the apoptotic rate showed tendency to ascend, with the increasing of multiplicity of infection and infection time. GES-1 cells were treated with multiplicity of infection of H. pylori at 100∶1 for 12, 24, 48 h, and the cell viability decreased to (80.57 ± 1.21)%, (70.04 ± 3.21)%, and (67.74 ± 2.91)%, while the apoptotic rate increased to (23.74 ± 1.71)%, (53.60 ± 1.87)%, and (70.67 ± 2.87)%. After co-cultured with 1.0 μg/mL Zuojin Formula for 24 h, the cell viability increased to (97.67 ± 1.04)%, and the apoptotic rate decreased to (31.04 ± 1.02)%, and the difference was statistically significant compared with the corresponding model group (P < 0.01). The results of Western blotting showed that the protein expression of Caspase-3 of all treatment groups was obviously decreased. Based on the morphological point of view, this result was further verified. Conclusion: GES-1 infected by H. pylori NCTC11637 can inhibit cell proliferation and induce apoptosis, while ZuoJinFang could protect GES-1 cells from cell damage.