Allicin Inhibits The Proliferation,Migration And Angiogenesis Of Vascular Endothelial Cells In Epidural Fibrosis

Objective: To investigate the effects of allicin on the proliferation,migration and angiogenesis of Rat vascular endothelial cells(RVES),and to explore the influencing mechanism of allicin on epidural fibrosis.Methods: According to the results of preliminary experiments,RVECs were divided into control group(0mg/L),low concentration group(25mg/L),medium concentration group(50mg/L)and high concentration group(100mg/L).The morphology,viability,migration rate,cell cycle,apoptosis rate and cell lumen formation ability were measured using fluorescence microscope,AO/EB double staining Annexin V-FITC double staining,PI/RNase staining,scratch assay and Transwell experiments test.Western Blot was used to measure the protein expression level of JAK2,STAT3,p-STAT3,PCNA,Bax and Bcl-2 protein.The experimental rats were grouped by random number method,and 36 adult male SD rats were divided into three groups,namely sham operation group,control group and allicin group.The rats in each group were treated according to the group.On the 28 th day after the operation,the rats were killed by overdose anesthesia.The epidural scars of each group were observed.The rat spinal canal area specimens and tissue sections were made.The sections were stained with HE staining and Masson staining.The number of fibroblasts,the number of blood vessels and the density of collagen in each group of stained sections were analyzed under an optical microscope.The animal hindlimb motor function score was used to evaluate the safety of the drug.One-way analysis of variance was used among multiple groups.Results:(1)compared with control group,the vitality,cell migration and lumen formation ability of rat vascular endothelial cells were decreased with allicin concentration increasing(p<0.01).(2)compared with the control group,the G1 phase and S phase in each rat vascular endothelial cells groups were decreased(p < 0.01),while in G2 phase was increased(p<0.01),cells were block in G2 / M phase.In addition,the apoptosis rate of vascular endothelial cells in allicin groups was increased with allicin concentration increasing(p<0.01).(3)Compared with the control group,the protein expression levels of JAK2,STAT3,p-STAT3,PCNA and Bcl-2 in rats vascular endothelial cells on allicin groups were down-regulated(p<0.01),while the protein expression of Bax was up-regulated(p<0.01),the ratio of p-STAT3/STAT3 was decreased(p<0.01),and the ratio of Bax/Bcl-2 was increased(p<0.01).(4)there were no death,infection or abnormal gait occurred in all the experimental animals.Laminectomy was not performed in the sham group,but for the two groups of rats underwent laminectomy,dense scar tissue firmly adhered to the dura could be seen in the epidural area of the control group.In contrast,there was a clear gap between the epidural scar and the dura in the allicin group,and the number of capillaries,collagen density and fibroblast density were significantly reduced(p<0.01).Conclusions: Allicin can inhibit the proliferation,migration and lumen formation of rat vascular endothelial cells,promote the apoptosis of rat vascular endothelial cells and inhibit the rat vascular endothelial cells in the G2/ M phase,the mechanism of which may be related to the inhibition of the JAK2/ STAT3 signaling pathway.In addition,animal studies showed that allicin effectively reduced collagen,vascular density,and fibroblast count in scar tissue,and improved epidural fibrosis after laminectomy.