Observed The Yanxuesan Multiplexing The Recipe Fiber Cells With Serum Pharmacology Method

ObjectiveThe experiment was performed for evaluating the effect of the TCM decoction named YanXueSan (YXS) on proliferation and apoptosis of NIH/3T3 cell by serum pharmacological method in vitro and compare to control group. To investigate the mechanisms of YXS which was composed according to TCM method of QuYuHuaTan(QYHT) in inhibiting proliferative vitreoretinopathy(PVR).To prove the effect of TCM method of QYHT by using cell test. To provide the scientific accordance and theory indication for the clinical using and developing medicine to prevent PVR. Methods1. Cell cultured: NIH/3T3 cell (mouse embryo fibroblast) was cultured in vitro by useing cell culture technology.2. Decoction prepared: YXS was prepared and preserved in 0℃ after being poached, alcoholized and settled, being concentrated to 2g/ml and filtrated, and being got ride of bacteria.3. Serum prepared: The control serum and medicated serum each was prepared after given the normal rabbits of 0.9%natriumchloride solution and YXS through gastrotavage for 3 days by serum pharmacological method.4. Groups separated: The log phase growth NIH/3T3 cell were divided into 10 groups: medicated serum20%,10%,5%concentration groups. Control serum20%,10%, 5%concentration groups. TCM decoction YXS 25mg/ml, 12. 5mg/ml, 6. 25mg/ml concentration groups. control group.5. Test: The growth cell morphological changes, collagen production and apoptosis of NIH/3T3 cell were tested by methyl thiazolyl tetrazolium (MTT) colorimetry, fluorescence dye, hydroxyproline reagcuat and flow cytometry each. Results1. Fibroblast proliferation inhibited: The proliferation effect of NIH/3T3 cell was inhibited significantly and dose-dependently after exposed to 3 different concentration YXS and medicated serum. When compared with the control group , YXS and 20%,10% medicated serum had significant difference (P < 0.01). 5% medicated serum had no significant difference (P>0.05).The control serum group could not inhibit NIH/3T3 cell proliferation (P > 0.05).2. Collagen production reduced: The soluble collagen production in culture medium was reduced significantly and dose-dependently after exposed to 3 different concentration YXS and medicated serum. When compared with the control group , YXS and 20%,10% medicated serum had significant difference (P < 0.01). 5% medicated serum had no significant difference (P>0.05). The control serum group could not reduce the soluble collagen production in culture medium (P > 0.05).3. Cell cycle controlled: The growth of NIH/3T3 cell was controlled significantly and does-dependently in G0/G1 stage after exposed to 3 different concentration YXSand medicated serum. When compared with the control group , YXS and 20%,10% medicated serum had significant difference (P < 0.01). 5% medicated serum had no significant difference (P > 0.05). The control serum group could not inhibit NIH/3T3 cell cycle (P > 0.05).4. Cell apoptosis premoted: The typical apoptosis characteristic morphological changes were found after exposed to YXS and medicated serum. The apoptosis rate increased significantly and dose-dependently. When compared with the control group , YXS and 20%,10% medicated serum had significant difference (P<0.01,P<0.05). 5% medicated serum had no significant difference (P > 0.05).The control serum group could not induce NIH/3T3 cell to apoptosis (P > 0.05). ConclusionYXS could prevent PVR by inhibiting NIH/3T3 cell proliferation, reducing cell collagen production, controlling cell cycle in G0/G1 stage, and inducing cell to apoptosis.