| Objective:To establish the model of SD rats full-thickness skin autograft. Observing the concentration of substance p(SP) and the numbers and distribution of epidermal stem cells in the skin during the take process after full-thickness skin autograft,observing the change of wound surface at the same time. To analyze the relationship between the change of SP and nerve fibers'wound-repair process during the process after skin autograft, and the relationship between SP,ESCs and wound healing, then try to study the role of SP in the take process of full-thickness. These results provided theoretical basis and experiment evidence for improving the quality of full-thickness skin autograft in practices.Methods:Sixty-three masculine SD rats, which were two to three months old, were randomly divided into seven groups: group D0(control group of normal skin), group D1(one day after operation), group D3(three days after operation), group D7(seven days after operation), group D14(fourteen days after operation), group D21(twenty-one days after operation), group D30(thirty days after operation). From group D1 to group D30, each rat was made full-thickness skin autograft model on back about 1.5cm×1.5??quare. At the different six times after operation, samples were cutted in each rat. In the group D0, we cut the full-thickness skin tissue about 1.0cm×1.0cm square from the back of each rat but do not establish the model of transplantation. 1.the concentration of SP by using radio immune analysis. 2.By the method of S-P immunohistochemistry, integrinβ1 and P63 were employed as the first antibody to demonstrate the epidermal stem cells. we compared the difference of the results between group D0 and other groups, and the difference of the results between each experiment group. The data were analyzed by SPSS13.0.Results:1. The full-thickness skin autograft on each rat was ideal; And there were not obvious infections and necroses in naked eye. But there were significant differences among different groups from optical observation.2. The results of radio immune analysis were: the concentration of SP reduced at the beginning. About the time of three days after operation, the concentration reached the minium in a month. Then the concentration began to raise. About thirty days after operation, the concentration still not reached the level that before operation. The concentration of SP in the D0 group was (20.88±1.25)pg/mg; the concentration of SP in the D1 group was(10.82±0.99)pg/mg; the concentration of SP in the D3 group was(4.08±1.18)pg/mg; the concentration of SP in the D7 group was (7.50±1.05)pg/mg; the concentration of SP in the D14 group was (11.80±1.39)pg/mg; the concentration of SP in the D21 group was(15.19±0.80)pg/mg; the concentration of SP in the D30 group was(16.76±1.33)pg/mg. Through comparing the difference between group D0 and other groups, we found that the results showed significantly statistical disparity(P<0.05). Thr?? comparing the difference between each experiment group, we found that the difference between group D1 and group D14 did not show significantly statistical disparity(P=0.078﹥0.05),but the difference between other experiment groups showed significantly statistical disparity(P<0.05).3. The results S-P immunohistochemistry were: The positive cells ratio(PCR%) of the two biomarks of epidermal stem cell reduced at the beginning. About the time of three days after operation, the PCR reached the minimum. Then the PCR began to raise; About seven days after operation, the PCR reached or overtook that before operation; Around twenty-one days, the PCR reached the maximum; But after then, the PCR began to reduce again; We still found the PCR above the level before operation on one month after operation. The PCR of P63 is higher than that of integrinβ1 on the immunohistochemistry samples which came from the same sample of each group. (1) integrinβ1: The PCR of integrinβ1 in group D0 were(18.11±1.05)%; The PCR of integrinβ1 in group D1 were (15.67±0.87)%; The PCR of integrinβ1 in group D3 were(8.22±1.20)%;The PCR of integrinβ1 in group D7 were (17.67±1.22)%; The PCR of integrinβ1 in group D14 were(19.89±1.17)%; The PCR of integrinβ1 in group D21 were(26.44±1.01)%; The PCR of integrinβ1 in group D30 were(20.11±1.27)%. Through comparing the difference between group D0 and other groups, we found that the difference between group D0 and group D7 did not show significantly statistical disparity(P=0.905﹥0.05),but the difference between group D0 and other experiment groups showed significantly statistical disparity(P<0.05). Through comparing the difference between each experiment group, we found that the difference between group D14 and group D30 did not show significantly statistical disparity(P=0.676﹥0.05),but the difference between other experiment groups showed significantly statistical disparity(P<0.05).(2)P63: The PCR of P63 in group D0 were (28.22±0.97)%; The PCR of P63 in group D1 were(25.56±1.13)%; The PCR of P63 in group D3 were(12.89±1.05)%; The PCR of P63 in group D7 were(28.78±1.09)%; The PCR of P63 in group D14 were(30.67±1.32)%; The PCR of P63 in group D21 were (37.78±1.39)%??e PCR of P63 in group D30 were(30.11±1.45)%. Through comparing the difference between group D0 and other groups, we found that the difference between group D0 and group D7 did not show significantly statistical disparity(P=0.838﹥0.05),but the difference between group D0 and other experiment groups showed significantly statistical disparity(P<0.05). Through comparing the difference between each experiment group, we found that the difference between group D14 and group D30 did not show significantly statistical disparity(P=0.336﹥0.05),but the difference between other experiment groups showed significantly statistical disparity(P<0.05).4. The positive cells were founded on the basal layer of epidermis, and at the folliculus pili bulge in the immunohistochemistry samples on the group D0. While on the group D1, we founded positive cells in all layers of epidermis. With the time going on, the degree became more and more serious. Beginning from group D21, we found the distribution of positive cells went back to the pattern before operation. Conclutions:1. During the take process of full-thickness skin autograft, the concentration of SP reduced at the beginning. And with the establishment of sensory nerve, the concentration of SP raised gradually. The change rules of substance p responsed the wound-repair process of sensory nerve.2. During the take process of full-thickness skin autograft, the ratio of epidermal stem cells reduced at the beginning. With the establishment of blood circulation and sensory nerve, the ratio of epidermal stem cells raised gradually.3. The distribution of epidermal stem cells during the take process of full-thickness skin autograft became disorder(found in all layers of epidermis) gradually; From the twenty-first day, it trended to the rule of normal (concentrated mainly in the basal layer of epidermis).4. According to the change rules of substance p and epidermal stem cells during the take process of full-thickness skin autograft, and with the change of wound surface, we can concluted that SP and epidermal stem cells have closed relation with wound healing. And substance p is a important substance duiring the take process of full-thickness skin autograft.
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