Effects on Recombinant Human Interleukin-10 of IL-2, IFN-γ, GCs, T Cell Apoptosis in Transplantation of the Rabbit Skin
|School||Zunyi Medical College,|
|Keywords||recombination human interleukin-10 skin graft cytokines T cell apoptosis glucocorticoids|
Objective:To observe the rabbit skin graft rejection and the change of IL-2,IFNγ,GCs,T cell apoptosis in peripheral blood after the injection of recombination human interleukin-10, to explore the mechanism of recombination human interleukin-10 in transplantation of the rabbits skin..Methods:(1)36 rabbits were divided into two groups which were receptors and donors, each donor rabbit preserve skin for one receptor,skin transplants on the back. They were randomly divided into six groups:rhIL-10 group (low-dose 5μg/kg/d, high-dose 10μg/kg/d), CsA positive control group (low-dose 5mg/kg/d, high-doses of 10mg/kg/d), rhIL-10 + CsA joint group (5μg/kg/d + 5mg/kg/d), NS negative control group (1ml/d).And each group have 3 rabbits.All groups were intramuscular injected drugs 10 days started from one day before the surgery of skin transplantation. (2)Observation the situation of grafted skin after the operation, record rejection time and survival time of the grafted skin; (3) Each group of rabbits’peripheral blood were collected at 1 day before surgey and 1thday,4thday,7thday,14thday,21thday,28thday after the operation.Use enzyme-linked immunosobent assay(ELISA) to test the chrange of IL-2, IFN-γ, GCS in peripheral blood and analyzed by flow cytometry(FCM) to test the change of the T cell apoptosis rate after surgey 1thday,4thday,7thday,14thday.Results:(1) General condition of rabbits after the skin graft:the rabbits of high-dose of CsA group had a poor diet and mental state and hair loss phenonmenon; however,the other groups didn’t appear such phenomenon. (2) Skin graft beginning rejection time and survivial time:Compared with the NS negative group, skin of the all medicine-consumption group of rabbits began to appear rejection time and survivial time are delayed (p<0.05). Compared with the CsA positive control group, skin of rhIL-10 group with corresponding doses’survivial time have no significant difference (p>0.05). rhIL-10 + CsA group have no significant difference (p>0.05). Compared with the low dose of IL-10 group, rhIL- 10 + CsA group’s skin began to rejection reaction time and live time are delayed (p<0.05). (3)The result of IL-2 and INF-γtest indicate that,Compared with the NS negative group, the other groups of IL-2 and INF-γ’gamma level is in different degree lower (p<0.05). Compared with the CsA positive control group, rhIL-10 with corresponding doses’IL-2 and INF-γhave no significant difference (p> 0.05). Compared with the high-dose CsA positive control group, rhIL-10+CsA group of serum IL-2 and INF-γ’slevel with no obvious difference (p> 0.05). Compared with the low-dose of IL-10 group and low-dose of CsA group, rhIL-10+CsA group of serum IL-2 and INF-γ’s level have significant difference (p<0.05). (4)T cell apoptosis rate and glucocorticoids tests showed that, Compared with the NS negative group, The other groups of T cell apoptosis rate and glucocorticoids’level is higher in different degree (p<0.05). Compared with the CsA positive control group, T cell apoptosis rate and glucocorticoids’ level of rhIL-10 with corresponding doses have no significant difference (p>0.05).Conclusion:(1) This experiments showed that the rhIL-10 inhibited the rejection on allogeneic skin graft, delayed the time of skin graft rejection, extended the survival time of grafted skin,and had a synergistic effect with CsA. (2) The mechanism of rhIL-10 may be inhibit IL-2 and INF-γ, promoting T cell apoptosis and glucocorticoids secretion to realize immunosuppressive and inducing immune tolerance.