The Research On Regulatory Mechanism Of Bangci Electroacupuncture In Promoting Wound Healing For Rats With Bedsore Through P38MAPK Signaling Pathway

Objective:To investigate the effects of Bangci ElectroAcupuncture(EA)on local skin blood flow perfusion in.rats with bedsore,to clarify the effects of EA in promoting wound healing;To investigate the effects of key factors of p38MAPK signaling pathway in wound tissue in rats with bedsore and to discuss the mechanism of EA in wound healing.Methods:fixing SD female rats with device on the leg near the knee joint bone and with ischemia-reperfusion injury the bedsore model was set up,which is established by forming of bedsore Ⅱ phase.SD rats were randomly divided into 5 groups:normal group,model group,the ElectricAcupuncture group,model inhabitor group and EA inhibitor group.Then the rats in any of a group according to the intervention timepoints,1d,3d,5d,7d and 9d were randomly divided into 5 sub-groups,which amount to 6.Normal group received the same amount of normal saline as SB203580;Model group received conventional iodine therapy after normal saline;after the model was built,in EA group the intervention was conducted after normal saline.Model inhibitor group received SB203580(5mg/kg)as p38 inhibitor,and then conventional iodine therapy.EA inhibitor group received SB203580 as p38MAPK inhibitor,conventional iodine therapy concurrently,and then Electric Acupuncture therapy at last.The method of EA is:to evaluate model of rats on the second day after the model and on the third day after the model successfully was established,the rats were fixed on the control console.Two needles were punctured in the skin,one vertically in the wound center and the other obliquely in the normal skin 0.5 cm away from the wounded skin edge,without any needling techniques,then micro-current electrical stimulation apparatus was connected.We put the positive pole on the handle which is on the wound center and the negative pole on the handle which is on the wounded skin edg with 500μA,0.5Hz for 30min,once a day.The planimeter was used to measure the rate of wound healing of the rats on different timepoint,the healing time was investigated too.The quantity of blood perfusion in bedsore organization was monitored by PeriCam PSI system at different timepoints for different groups.Immunohistochemistry method was used to detect the expression of p3 8MAPK and protein HSP27 in skin tissue near the wound for different groups,and the The number of CD34 microvascular on different timepoints.Western Blot method was used to detect the expression of p38MAPK and protein HSP27.Realtime-PCR method was used to detect the expression of p3 8mRNA and HSP27mRNA in skin tissue at different groups on different timepoints.Results:First part of the research:The results show that there were significant difference between Bangci EA group and model group on the wound healing rate,EA group was better than model group.There were significant difference between Bangci EA group and model group on the healing time.After application of specific inhibitor SB203580,the healing time was extended and the rate of wound healing declined.The second part of research:The results of the quantity of blood perfusion show that compared with normal group,the other four groups were significant better(P<0.05)at 1d;there were no Significant difference among them at 9d(P>0.05).Quantity of blood perfusion of EA group and model group at ld,3d,5d,7d,9d show a downward trend.The treatment markedly increased quantity of blood perfusion in EA group at every timepoint(P<0.05).The same situation hadn’t happened in the model group(P>0.05)After subcutaneous injection of inhibitor SB203580,there were no significant difference after the treatment either for EA group or the model group.The third part of research:results of immunohistochemical method show that the number of positive cells of p38MAPK and HSP27 in model group and EA group increased significantly compared with normal group at Id.At timepoints of 3d,5d,7d and 9d,the positive cells of p38MAPK and HSP27 in EA group decreased gradually;At timepoints of 3d,the positive cells of p38MAPK and HSP27 in model group achieved its peak,and then decreased at 5d,7d and 9d;the peak for model group achieved at 3d and the peak for EA group at 1d.After subcutaneous injection of inhibitor SB203580,p38MAPK and HSP27 protein expression decreased in either EA inhibitor group or model inhabitor group compared with EA group and model group,the The number of CD34 microvascular achieved its peak at 5d in EA group and at 7d in model group.The fourth part of the research:results of Western Blot show that the number of p38MAPK and HSP27 in model group and EA group increased significantly compared with normal group at ld(P<0.05).At timepoints of 3d,5d,7d and 9d,the positive cells of p38MAPK and HSP27 in EA group decreased gradually;At timepoints of 3d,the positive cells of p38MAPK and HSP27 in model group achieved its peak,and then decreased at 5d,7d and 9d;the peak for model group achieved at3d and the peak for EA group at Id.After subcutaneous injection of inhibitor SB203580,p38MAPK and HSP27 protein expression decreased in either EA inhibitor group or model inhabitor group compared with EA group and model group.Realtime-PCR results:results of immunohistochemical method show that the number of p38MAPKmRNA and HSP27mRNA in model group and EA group increased significantly compared with normal group.At timepoints of 3d,5d,7d and 9d,the positive cells of p38MAPKmRNA and HSP27mRNA in EA group decreased gradually;At timepoints of 3d,the positive cells of p38MAPKmRNA and HSP27mRNA in model group achieved its peak,and then decreased at 5d,7d and 9d;the peak for model group achieved at 3d and the peak for EA group at 1d.After subcutaneous injection of inhibitor SB203580,p38MAPKmRNA and HSP27mRNA protein expression decreased in either EA inhibitor group or model inhabitor group compared with EA group and model group.Conclusion:1 EA effectively promoted the wound-healing for bedsore of rats and the healing time equired was shorten.2 EA effectively markedly increased quantity of blood perfusion for bedsore of rats and involved in restoration of wounds.3 p38MAPK signal transduction pathways involved in and promote the wound-healing for bedsore of rats,with the application of p38MAPK signal transduction pathway inhibitor SB203580,wound healing time will be extended.4 EA can activate p38MAPK signaling pathways,promote p38MAPK and HSP27 protein and gene expression,and thus play a role in promoting wound healing;application of specific inhibitor SB203580,the reaction was inhibited remarkablely.5 p38MAPK signal pathway is the important target of Bangci electro-acupuncture and is a possible Mechanism for wound-healing for bedsore.