Influence of matrine on angiogenesis factors of glioma C6 cells
Objective To explore the influence of matrine on angiogenesis factors of glioma C6 cells under hypoxia.Methods The glioma C6 cells were cultured to logarithmic phase,and were added 0.8,1.0,1.2,1.4,1.6,1.8 and 1.6 mg/mL matrine solution.The inhibiting rate of glioma C6 cells were detected by MTT test.The glioma C6 cells in ogarithmie phase were randomly divided into CoCl2 normal group,normal oxygen group,CoCl2 matrine group,and normal oxygen matrine group.CoCl2 normal group and CoCl2 matrine group were cultured by CoCl2 nutrient solution,and normal oxygen group and nomal oxygen matrine group were cultured by DMEM.Inverted microscope was adopted to observe the cytomorphological changes of all groups.Western blot was adopted to detect the expressions of angiogenesis factors such as hypoxia inducible factor-1alpha (HIF-1α),vascular endothelial growth factor (VEGF) and tissue inhibitor of metalloproteinase 3 (TIMP-3).Results After being cultured with CoCl2 nutrient solution,matrine had obvious inhibiting effect on glioma C6 cells,and the inhibiting rate increased obviously with the increase of matrine concentrations (24 h:R2 =0.9897,IC50 =1.468 g/L).The cells were long and narrow,fusiform-shaped and adherent growing in normal oxygen group,in three-dimensional distribution in CoCl2 normal group,turned small and round,and finally pieces in normal oxygen matrine group,and slightly turned small and round and finally pieces in CoCl2 matrine group.The levels of HIF 1α (1.660±0.171,1.331±0.112),VEGF (1.161±0.020,0.932±0.016) and TIMP 3 (1.464±0.047,1.785±0.043) in CoC12 normal group and CoC12 matrine group were significantly higher than those in normal oxygen group (1.092±0.082,0.784±0.042,0.603±0.052) and normal oxygen matrine group (0.734±0.080,0.637±0.022,1.203±0.069) (P＜0.05),the levels of HIF-1α and VEGF were significantly lower and the level of TIMP-3 was significantly higher in CoC12 matrine group than those in CoC12 normal group (P＜0.05),and the levels of HIF-1α and VEGF were significantly lower and the level of TIMP-3 was significantly higher in normal oxygen matrine group than those in normal oxygen group (P＜0.05).Conclusion Under hypoxia,matrine could inhibit the proliferation of glioma C6 cells,down-regulate VEGF,and up-regulate TIMP-3,which may be correlated with the inhibition of angiogenesis.
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