| Objective: to discuss skin regeneration medical technology(MEBT/MEBO)promoting chronic ulcer healing mechanism of skin wound in rat from p38 MAPK signaling pathway.Methods:(1)the 40 SPF SD male rats which were 10~12 weeks old,the weight between 220g~250g,adaptability to feed a week were modeled in chronic ulcer model.In accordance with the principle of random sampling as blank group,10 rats blank group rats for full-thickness skin excision,only no longer drug intervention.Injection of hydrocortisone acetate to form the remaining 30 rats chronic wound model,were randomly divided into model group,MEBO group and the Beifuji group,each group of 10 only.All groups were using drug treatment on the second day.Before switching 2 times a day,all groups use 1/5000 Nitrofurazone solution to clean the wound.In 3 days,7 days,14 days of three time points,observing the wound healing time in each group,as well as calculating the healing rate.(2)as well as the requirement of 80 rats,in accordance with the principle of random sampling 20 rats as blank group,which for full-thickness skin excision,only no longer drug intervention.Remaining 60 injection of hydrocortisone acetate formation in rats with chronic wound model,were randomly divided into model group,MEBO group and the Beifuji group,each group of 20.All groups were using drug treatment on the second day.Before switching 2 times a day,all groups use 1/5000 Nitrofurazone solution to clean the wound.Five rats in each group were choosed to extract the tissue samples in the 3d,7d,14 d at random.Part of the tissue samples were fixed with 10% Formalin for pathological,the other part of the tissue samples were put in liquid nitrogen tank immediately,quickly moved to-80 ?cryogenic refrigerator later.Treating specimens were collected for subsequent Western blotting,Rt-pcr and ELISA methods.Results:(1)wound general situation: observed the growth of granulation tissue after making models,the recovery of blank group wound was best,MEBO group and the Beifuji complex economic group were significantly better than model group;(2)wound healing time: the healing time of the blank group was shortest(13.40±0.74d),which was a significant difference with the model group(P < 0.01),and model group was needed for the longest time(22.19±2.09d),which was a significant difference with the other three groups(F=141.900,P < 0.01);(3)wound healing rate: in 3d,7d,14 d,the healing rate of the blank group was significantly higher than the model group(P<0.05),as well as the MEBO group and Beifuji group compared with the model group,in 3d,14 d,there was no difference between the MEBO group and Beifuji group(P>0.05),but in 7d,the difference was statistically significant(P<0.05);(4)Western blotting:(1)in the whole process of wound healing,protein expression on the third day after making model preparation was the highest,followed by gradually decreased(P<0.01).the levels of p38,MK2 total quantity of protein expression in each group at 3d,7d,14 d the same time point,when there was no statistically significant difference between groups(P>0.05);(2)in the early trauma,the protein expression level of p-p38,p-MK2 in model group,MEBO group,the Beifuji group wound tissue were no statistically significant difference(P>0.05);at 7d,the differences between groups began to clear,the amount of protein expression in the blank group and MEBO group began a significant reduction,model group decline was not obvious,the difference between the each two groups was statistically significant(P<0.05);at 14 d,compared with the other three groups,model group was still in the high level expression(P<0.05);(5)Rt-pcr: in the process of the whole experiment,compared with the blank group,the quantity of p38 m RNA,MK2 m RNA expression in MEBO group,the Beifuji group and model group at 3d were the most,then continued to fall;at 3d,there was no statistically significant difference between each other(P>0.05),and at 7d,MEBO group and Beifuji group complex expression decreased obviously,the deference between the two group was statistically significant(P<0.05);at 14 d,the expression of MEBO group,Beifuji group complexly volume continued to decline,the two group has no statistically significant difference(P>0.05),but compared with the model group,the deference was statistically significant(P < 0.05),the model group was still in the high expression.(6)ELISA: in the process of the experiment,the quantity of TNF-?,IL-6 levels in the chronic wound tissue were declining over time,but with MEBO group had the greatest reduction,the model group decreased the most slowly.At 3d,the expression quantity of TNF-?,IL-6 in model group,MEBO group and Beifuji group were higher than that of blank group(P<0.05);at 14 d,the quantity of expression in the model group was still higher than the other three groups as well as at 7d(P<0.05).Conclusions:(1)MEBT/MEBO can obviously shorten the chronic wound healing time,improve the healing rate.(2)MEBT/MEBO can reduce the quantity of p-p38,p-MK2,p38 m RNA,MK2 m RNA expression in the chronic wound tissue.(3)MEBT/MEBO can reduce the guantity of TNF-?,IL-6 in the chronic wound tissue,and reduce local inflammation of the wound.On the whole: MEBT/MEBO can promoting the chronic wound healing,one of its mechanisms may by lowering the level of p-p38,p-MK2 protein expression through p38 MAPK signaling pathway which was excessive activated,blocking part related to signal transduction,reducing inflammation reaction,regulating the cellular immunity and promoting the process of rational proliferation of granulation tissue. |
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