The Potential Protective Effect Of Procyanidins On Excessive Proliferation And Migration Of VSMCs Induced By PM2.5

Objective:In this study,the VSMCs of rats were used as an evaluation model for toxicity studies.The purpose of this study was to observe the potential protective effects of procyanidins on excessive proliferation and migration of VSMCs induced by PM2.5.Methods:Healthy male SD rats were selected with weighing 100-120 g,and their thoracic and abdominal aorta were isolated under sterile environment,and primary VSMCs were cultured by tissue patch method.This experiment was divided into 5 groups: control group,PM2.5 groups(final concentration of 100?g/m L),and low,medium,high procyanidins group(final concentration of 5?mol/m L,10?mol/m L and 20?mol/m L).The ROS fluorescence intensity of different groups were observed with DCFH-DA.The cell proliferation of each group was detected by CCK-8 kits and Ed U cell proliferation kits.The cell migration of each group was tested with Transwell chamber and cell scratch wound assay.Western blot was used to detect the expression levels of MMP9,PCNA,Akt,p-Akt,GSK-3?,p-GSK3? and ?-actin in VSMCs.The m RNA levels of MMP9,PCNA and ?-actin were tested by RT-PCR.Results:1.Compared with the control group,the ROS fluorescence intensity of PM2.5 group was significantly increased.The ROS fluorescence intensity in the different procyanidins was decreased compared with PM2.5 group.2.According to the results of CCK-8 kits and Ed U cell proliferation kits,compared with the control group,the cell proliferation of PM2.5 group was significantly increased.The cell proliferation of different procyanidins groups was decreased compared with PM2.5 group.3.The results of Transwell chamber and cell scratch wound assay showed that,both in the longitudinal and transverse directions,the number and distance of cell migration in the PM2.5 group were significantly increased compared with the control group,while the number and distance of cell migration in the different procyanidins were reduced than the PM2.5 group.4.The changes in PCNA and MMP9 protein and m RNA levels as the indicators of cell proliferation and migration: compared with the control group,the expressions of PCNA and MMP9 were increased significantly in PM2.5 group.Compared with PM2.5 group,the expressions of PCNA and MMP9 were decreased in the different procyanidins groups.5.The activity of the PI3K/Akt signaling pathway was main through PM2.5,which could cause Akt to phosphorylate,and up-regulate the level of p-Akt.The up-regulated of p-Akt can negatively regulate the phosphorylation of GSK-3?,and the level of p-GSK3? decreased.Compared with PM2.5 group,the levels of p-Akt were down-regulated and the levels of p-GSK3? were up-regulated in the different procyanidins groups,which were statistically significant(P<0.05).Conclusions:procyanidins can potential protective the excessive proliferation and migration of VSMCs induced by PM2.5,and the protective mechanism may be related to the PI3K/Akt signaling pathway activated by procyanidins.