| Objective:Using the ance inflammatory rat model as experimental subjects to observe the gross changes,histopathological features and the expression of TNF-? and MMP-2,and explore the effect of h AMSC supernatant on the inflammatory model of acne rat.Methods:A total of 49 SD rats,weighing 250 ± 30 g,were randomly divided into 7 groups(A,B,C,D,E,F and G),each group of 7.Group A was blank control group,Group B was saline injected group,group C was acne model group,group D was acne model untreated group,group E was isotretinoin erythromycin gel treatment group,group F was h AMSC supernatant rubbing treatment group,Group G was injected with h AMSC supernatant treatment;group A was remained unhandled and sacrificed 7 days later;group B was injected 50?L saline into the central of two ears of SD rats,a total of 1 time and sacrificed7 days later;group C,D,E,F,G were injected 50?L propionibacterium acnes into the central of two ears of SD rats,a total of 1 time,established acne model.Group C was sacrificed after 7 days of modeling.Group D was sacrificed after observed 4 weeks.E,F,G was sacrificed after treatment.Two days after the modeling,the auricular swelled part was inoculated on the blood plate by aseptic needles and incubated in anaerobic tank for 48 hours to observe the colony morphology.The aseptic needles were used to pick the colony,smear and observe the bacterial morphology under a light microscope.During the experiment,seven groups of animals were fed normally,and observe the changes after the injection.The rats in each group were killed after the experiment.The tissue of auricle was stained with HE to observe the pathological changes of auricle tissue and the expression of TNF-? and MMP-2 protein in auricle tissue was detected by immunohistochemical method.Results:1.Microbiological examination: Microbiological examination of swollen auricular tissue with a sterile needle after modeling was confirmed as Propionibacterium acnes infection.2.Modeling results to judge: 1)Gross observation revealed that there was no significant changes were observed in group A;in group B the ears of the rats became swelling,24 hours later,the swelling subsided;while in group C the ears swelling and visible papules.2)The results of histopathology showed that there was no obvious inflammatory cells were found in group A and B;in group C,hyperkeratotis,inflammatory cells infiltrated,part of the hair follicle area expanded,the structure of subcutaneous tissue disappeared.3)Immunohistochemical results of TNF-? and MMP-2 showed that weak positive protein was seen in group A and B,strong positive expression of protein was observed in group C,and positive protein was up-regulated in group C compared with group A and B,has statistical significane(P <0.05).3.Acne model untreated and treated group judgment: 1)Gross observation revealed that the auricular papules increased significantly in group D,and in group E,F and G,the auricular papules basically subsided.2)The results of histopathology showed that the results of group D were the same as group C;the sebaceous follicle had narrowed with a few inflammatory cells scattered in E,F and G group.3)Immunohistochemical results of TNF-?and MMP-2 showed that there was no significant difference between the D group and the C group(P> 0.05).The positive expression of protein in group E,F and G was down-regulated compared with group C and D,has statistical significane(P <0.05).There was no significant difference between the E,F,G group and the A,B group(P> 0.05).There was no significant difference between the E,F and G groups(P > 0.05).Conclusion:1.h AMSCs culture supernatant can reduce auricular inflammation in rats,and the mechanism may be that IL-10 and TIMP can inhibit the expression of TNF-? and MMP-2 in the supernatant,thereby reducing the inflammatory response.2.There is no significant difference in curative effect between the treatment group and the injection group in the supernatant of culture supernatant of h AMSCs culture. |
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