The Role Of Protein Kinase C-alpha In Recombinant Angiostatin Inhibition On The Rabbit Ears Wound Healing

Objective To research the roles of PKC-a in recombinant human angiostatin(K1-3) inhibition on wound healing and angiogenesis in the ventral skin of rabbit ears.Methods The wound healing model was built on the ventral skin of rabbit ears.The wounds were respectively treaed with PBS, recombinant human angiostatin, PMA, G56983, recombinant human angiostatin combined with PMA or Go6983once a day until epithelization(13-16days).Wound images were collected to calculate wound healing rates with Image-Pro Plus6.0software on3d,7d and11d.And wound healing time was recorded.Microvessel density(MVD) and the expression of PKC-α in wound healing process was detected by immunohistochemical staining with vascular endothelial cell-specific marker CD34and PKC-α polyclonal antibody.The wound healing time, the wound healing rates, the MVD and the expression of PKC-α were expressed with Mean±tandard difference (x±s),which were assessed by one-way ANOVA when the homogeneity of variance between groups and their further multiple comparison by LSD(least significant different) method, using the Kruskal-Wallis Test when heterogeneity of variance.Results1.The wound healing time in rh-AS group and Go6983group was longer than control group but PMA group was shortened compared with control group(P＜0.05).The wound healing time in rh-AS group was longer than rh-AS+PMA group,but shorter compared with rh-AS+Go6983group(P＜0.05).The wound healing time in PMA group was shorter in rh-AS+PMA group(P＜0.05).And the wound healing time in Go6983group was shorter compared with rh-AS+Go6983group(P＜0.05).2.There were differences on wound healing rates in every group at different time:Wound healing time on the3rd day after injury in every group was no significant difference(P＞0.05).On the7th and11th day after injury,the wound healing rates in rh-AS group and Go6983group were lower than control group,but PMA group was higher than control group(P＜0.05).Rh-AS group had lower wound healing rates than rh-AS+PMA group,and higher compared with rh-AS+Go6983group(P＜0.05).The wound healing rates in PMA group and Go6983group was higher than in rh-AS+PMA group and rh-AS+Go6983group,respectively(P＜0.05).3.Microvessel density:The microvessel density of rh-AS group, Go6983group and rh-AS+Go6983group was lower than control group on the any day after injury (P<0.05) whereas that of PMA group was higher than the other groups(P＜0.05). The microvessel density in every group declined with the wound epithelialization, but that in PMA group was higher than other groups(P＜0.05).4.The expression levels of PKC-α:PKC-α was widely expressed in all kinds of cells the wound such as fibroblasts,endothelial cells,epidermal cells. The expression of PKC-a and the proportion of positive cells(fibroblasts,endothelial cells,epidermal cells) increased in every group with the wound healing progress.On the same time, the expression levels of PKC-a in rh-AS group was no significant difference with control group(P＞0.05).The proportion of positive cells in PMA group or rh-AS+PMA group was higher than control group and rh-AS group.The expression levels of PKC-a in Go6983group and rh-AS+Go6983group was lower compared with control group or rh-AS group(P＜0.05).Conclusions1. The angiostatin might inhibit the angiogenesis in the wound healing and delays wound healing.2. PKC-a might not mediate the inhibition effects of angiostatin on the wound healing and angiogenesis.3. PMA might be conducive to the wound healing whereas Go6983might delay wound healing.