The Effects Of Gypenosides To The Proliferation Of Human Pterygium Fibroblasts

Objective:Pterygium is a common, multiple ocular surface disease and easy to recur. Gypenosides (Gyp) is the major components of gynostemma, a kind of cucurbitaceae. Many years of pharmacological researches from home and abroad showed that Gyp had anti-tumor and anti-oxidant effects, and also played roles in reducing lipids and blood glucose, preventing fibration in liver and kidney, and so on. These findings above empower Gyp to be a potential prevention and therapy method of pterygium. However the exact mechanism is unknown. The aim of this work was to study the effects of Gyp to the proliferation of human pterygium fibroblasts (HPF) initially.Methods: The tissues from pterygium surgerys were cultivated in vitro. After the primary culture and subculture, the third to sixth passage of HPFs were used for immunohistochemical staining with anti-vimentin antibodies, combining with morphological features, pterygium fibroblasts can be identified. HPFs were incubated with Gyp (25, 50, 100, 200, 400μg/ml) for 24, 48, 72 and 96 hours respectively. The negative control group without drugs and the control group with only culture solution were installed at the same time. The MTT method was used to evaluate the cell proliferation inhibitory rate and assay the biologic activities of Gyp in different time and different doses. Cell climbing films were processed with Gyp (0, 25, 50, 100, 200, 400μg/ml) for 24 hours, and immunocyte chemical method was used to analyze expressions of proliferating cell nuclear antigen (PCNA).Results: (1) The expression of vimentin in cultured cells in vitro was positive, pterygium fibroblasts were identified. (2) MTT results showed that Gyp can inhibit the proliferation of pterygium fibroblasts significantly when its concentration was not less than 25μg/ml (p<0.05). The proliferation inhibitory rate of Gyp on HPFs increased as incubation time (24-72 hours) and concentration (25-400μg/ml) increasing. When the intervention time was more than 72 hours, no significant increase in inhibition was found. (3) When the concentration ranged in 25-400μg/ml, Gyp can inhibit the expression of PCNA in fibroblast in a concentration-dependent manner (p<0.05).Conclusions: The inhibition effect of Gyp on HPFs was significant in vitro, and in a dose-and time-dependent manner when in a certain range of concentration and time.