The Effect Of Verapamil, Nimodipine, Total Saponins Of Panax Notoginseng On The Proliferation Of Rats Lens Epithelial Cells And The Synthesis Of Extracelluar Matrix

Objective: The purpose of these experiments was to study the effect of the calcium channel blockers-verapamil,nimodipine and total saponins of panax notoginseng on the proliferation of rat lens epithelial cells and the synthesis of extracelluar matrix and to provide basic methods and experimental basis for the mechanism of medication treatment of the after cataract .Methods: Eighty-new-born rats lens were obtained and cultured with digest and tissue culture method and 2-3 passage RLECs were placed in 96-well flat bottomed tissue culture plates and incubated for 24 hours. Different concentrations of Ver, Nim and Pns were added and the proliferation characteristics of the lens were measured by a rapid colorimetric assay(MTT assay) after separate culture for 24 hours,48 hours and 72 hours. The effect of expression of the three medications on the ECM-laminin and IV collagen was observed with immunocytochemical staining after being separately cultured for 24 hours ,48 hours and 48 hours. The section of eye balls of six adult SD rats and six new-born rats were obtained and the expression of lens IV collagen was observed with immunocytochemical staining.Results: 1. Rats lens epithelial proliferation rate was decreased respectively with the increase of the concentration of Ver,Nim,Pns and showed all remarkably decrease in 72 hours culture. No significant difference of rats lens epithelial proliferation rate was observed in Ver,Nim after culturing separately for 48 hours and 72 hours, while significant difference observed after culturing for 24 hours and 72 hours.Obvious difference was seen in using Pns after cuturing for 24 hours ,48 hours and 72 hours.2. With the increased concentration of Ver, Nim and Pns,the expression of rats lens epithelial laminin decreased and obvious difference was observed after culturing separately for 48 hours , 72 hours and 24 hours.3.In these experiments, no expression of IV collagen was found in both culture and rats eyes section of immunocytochemical staining and there is the expression of IV collage in the corneal tissue with section of rats eye balls.Conclusion: these experiments confirmed that Ver, Nim and Pns could inhibit the effects on RLECs, and it would be enhanced greatly as the concentration and the time increased. There was no significant difference among these three medications and they had the inhibitory effect on synthesis of epithelial cell laminin and no expression of the IV collagen was found.