Objective To investigate the mechanism of stromal vascular fraction-gel(SVF-gel)in the treatment of hypertrophic scar in rats.Methods SVF-gel was isolated and extracted from adipose tis-sue,and the rat hypertrophic scar model was established.A total of 12 scar model rats were divided into experimental group and control group(6 rats in each group)according to the principle of complete random control according to body weight and scar size,and 0.2 ml of SVF-gel prepared in advance was evenly in-jected into the scar,and the control group was injected with the same amount of normal saline.After 6 weeks of continuous injection,the area of the scar in the rats was observed.The scar tissue of rats was collected after operation,and the level of scar protein was detected.SVF-gel conditioned medium(SVF-CM)was prepared,and the effects of different concentrations of SVF-CM on fibroblast proliferation were observed by cell counting kit-8(CCK-8)experiment,and the optimal concentration and time were de-termined.Using the gene knockdown kit,the fibroblasts with transforming growth factor-β1(TGF-β1)gene knockdown were obtained,and following experimental groups were set up:group A[fibroblast(Fb)+NC small interfering RNA(siRNA)],group B(SVF-CM+NC siRNA),group C(SVF-CM+NC siRNA+periostin),group D(SVF-CM+TGF-β1 siRNA),group E(SVF-CM+TGF-β1 siRNA+Periostin).Western blotting was used to observe the expression of Periostin,TGF-β1,type Ⅰ and type Ⅲ collagen.Graphpad software was used to analyze the statistical results by t test.Results In vivo experiments,the scar area in the control group was greater than that in the experimental group[scar area:(21.18±2.53)mm2 vs.(7.49±1.31)mm2,t=10.75,P＜0.01;hematoxylin and eosin(HE)staining of the dermal scar area:(17.69±3.22)mm2 vs.(2.43±1.37)mm2,t=9.75,P＜0.01).Western blotting showed that the expression of Periostin and TGF-β1 was higher in the control group than in the experimental group(Periostin:0.91±0.06 vs.0.52±0.02,t=14.16,P＜0.01;TGF-β1:1.00±0.03 vs.0.52±0.06,t=15.46,P＜0.01).In vitro experiments,CCK-8 experiments showed that the proliferation ability of fibroblasts treated with 40％SVF-CM for 48 h was higher in the control group than in the experimental group[(100.00±6.95)％vs.(79.89±2.51)％,t=7.11,P＜0.01].Western blotting showed that af-ter TGF-β1 knockdown,the expression of Periostin,TGF-β1 protein,type Ⅰ and type Ⅲ collagen was higher in group B than in group D(Periostin:0.763±0.005 vs.0.602±0.004,t=44.24,P＜0.01;TGF-β1:0.694±0.026 vs.0.588±0.033,t=4.381,P＜0.05;Collagen Ⅰ:0.749±0.002 vs.0.472±0.007,t=63.51,P＜0.01;Collagen Ⅲ:0.752±0.019 vs.0.593±0.009,t=13.34,P＜0.01).When exogenous Periostin was added(group E),the expression of periostin and type Ⅰ and typeⅢ collagen was higher in group E than in group D(Periostin:0.826±0.010 vs.0.602±0.004,t=44.24,P＜0.01;Collagen Ⅰ:0.947±0.002 vs.0.472±0.007,t=109.40,P＜0.01;Collagen Ⅲ:0.924±0.019 vs.0.593±0.009,t=27.33,P＜0.01).Conclusion SVF-gel can inhibit TGF-β1/Smad signaling pathway,thereby reducing the expression of Periostin,inhibiting fibroblast proliferation,and reducing the production of collagen Ⅰ and Ⅲ to prevent the formation of hypertrophic scars.
NETL
NSTL
万方数据
