Effects of allicin on the biological behavior of vascular endothelial cells and epidural scar / 中华骨科杂志

ABSTRACT

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 (0 mg/L), low concentration group (25 mg/L), medium concentration group (50 mg/L) and high concentration group (100 mg/L). The morphology, viability, migration rate, cell cycle, apoptosis rate and cell lumen formation ability were measured using fluorescence microscope, AnnexinV-FITC double staining, PI/RN-asestaining, 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. Using random number method, 36 adult male SD rats were divided into sham operation group, saline group and allicin group, with 12 rats in each group. Hematoxylin-eosin (HE) staining, Masson staining and immunohistochemical staining were used to analysis the epidural fibrosis in each group.Results:With the increase of concentration of allicin, cell viability, cell migration and lumen formation ability significantly lower than that of control group ( P<0.05). With the increase of allicin concentration, the percentage of cells in the G1 and S phases gradually decreased ( P<0.05), the percentage of cells in the G2 phase and the apoptosis rate gradually increased ( P<0.05), and the cells were blocked in the G2/M phase. With the increase of allicin concentration, the protein expression levels of JAK2, STAT3, p-STAT3, PCNA and Bcl-2 were gradually down-regulated ( P<0.05), while the protein expression level of Bax was gradually up-regulated ( P<0.05), the ratio of p-STAT3/STAT3 was decreased ( P<0.05), and the ratio of Bax/Bcl-2 was increased ( P<0.05). There was no death, infection or abnormal gait in all the experimental animals. Dense scar tissue could be observed in the extradural area of the sham operation group and the epidural area of the control group, but there was obvious space between the epidural scar and the dura mater in the allicin group, and the density of collagen, the number of blood vessels, and the protein density of p-STAT3 were significantly lower than those in the control group ( P<0.05). Conclusion:Allicin inhibits angiogenesis and the severity of epidural scar after laminectomy, and the mechanism may be through inhibiting the JAK2/STAT3 signaling pathway of vascular endothelial cells.