Abstract:  Objective To observe and analyze the inhibitory effects of gentiopicroside (GPS) on the proliferation and migration abilities of human Tenon's capsule fibroblasts (HTCF) cultured in vitro. Design Experiment study. Participants Human Tenon's capsule fibroblasts. Methods Human Tenon's capsular tissue was obtained during the strabismus correction surgery at our hospital. HTCF were identified through immunofluorescence staining, and the well-cultivated third to fifth generation HTCF were utilized in the experiment. The drug groups were established with GPS concentrations of 5, 10, 20, 35, 45 and 50 μg/mL, respectively, and the negative control group with a GPS concentration of 0 μg/mL (NC group) was also set. CCK-8 assay was employed to measure the cell proliferation capabilities of HTCF in the various groups at 24, 48, 72 and 96 h. Cell scratch assay was used to examine the cell migration abilities of HTCF in various groups at 6, 12, 24 and 48 h. Main Outcome Measures The cell identification, cell proliferation amount and cell migration amount. Results Cultured cells grew well. The positive immunoreactivity for Vimentin and fibronectin (FN1) was observed. Specifically, Vimentin manifested as uniform red fluorescence and fibronectin as uniform green fluorescence within the cytoplasm, respectively, while the DAPI stains the nucleus with blue fluorescence. CCK-8 assay showed that significant differences in cell proliferation ability among HTCF incubated for 48, 72 and 96 h with various concentrations of GPS (F48h=14.947, F72h =25.683, F96h =131.392, all P＜0.001). The cell proliferation ability of HTCF in the GPS drug groups with concentrations of 35, 45 and 50 μg/mL was significantly reduced at 48, 72 and 96 h, there were significant differences compared with the NC group (all P＜0.01). The scratch assay results indicated that, in contrast to the NC group, no significant differences were found in the cell migration abilities of all GPS drug groups at 6, 12 and 24 h (all P＞0.05). At 30 h, the HTCF in the NC group, as well as those in the GPS drug groups with concentrations of 5, 10 and 20 μg/mL, had closed. Nevertheless, when compared with the NC group, the cell migration abilities in the GPS drug groups with concentrations of 35, 45 and 50 μg/mL were significantly decreased at 30 h, and all these differences were statistically significant (all P<0.001). Conclusion GPS at a concentration of 35 μg/mL significantly inhibited the proliferation and migration of HTCF. (Ophthalmol CHN, 2025, 34: 110-115)

Key words: gentiopicroside, human Tenon's capsule fibroblasts