摘要
目的探讨脂肪源性干细胞(ADSCs)通过抑制炎症对兔耳增生性瘢痕的治疗效果。方法选取20只清洁级新西兰大白兔为实验对象,取其中2只获取脂肪源性干细胞,将其余18只随机分为对照组(n=9)与治疗组(n=9)。建模成功后,对照组每点注射0.3 ml生理盐水;治疗组从瘢痕四周正常皮肤约0.4 cm处刺入至瘢痕中央,每点注射0.3 ml(1×10;个)ADSCs。分别于术后1、3、5周将每组3只动物处死取材,应用酶联免疫吸附实验(ELISA)检测白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的含量;应用实时荧光定量PCR检测Ⅰ型胶原(COLⅠ)及α-平滑肌肌动蛋白(α-SMA)的表达水平;通过HE染色检测并观察组织形态。结果与相应的同型对照比较,ADSCs表面抗原CD73(99.9%)和CD90(99.9%)呈阳性表达,而CD34(2.09%)和CD45(1.36%)呈阴性表达,证明脂肪源性干细胞提取成功。ELISA检测结果显示,与对照组相应时间点比较,ADSCs作用1、3、5周时,组织中IL-6的表达均显著下调,TNF-α的表达均被抑制,差异有统计学意义(P<0.05)。实时荧光定量PCR实验结果显示,与对照组相应时间点比较,ADSCs作用1、3、5周时,组织中COLⅠ和α-SMA的表达均显著下调,差异有统计学意义(P<0.05)。HE染色结果显示,ADSCs的添加可显著降低组织中炎性细胞聚集的现象,对照组胶原错乱排列的现象比较严重。结论ADSCs能够通过下调增生性瘢痕中炎症因子的表达,抑制COLⅠ和α-SMA的含量,进而发挥抑制增生性瘢痕的作用。
Objective To investigate the effect of adipose-derived stem cells(ADSCs)on rabbit ear hypertrophic scar by inhibiting inflammation.Methods Twenty clean New Zealand white rabbits were selected as experimental subjects.Fat-derived stem cells were obtained from 2 of them,and the other 18 rabbits were randomly divided into control group(n=9)and treatment group(n=9).After successful modeling,the control group was injected with 0.3 ml normal saline at each point.In the treatment group,0.3 ml ADSCs(1×10~5 PCS)were injected at each point from 0.4 centimeters of normal skin around the scar to the center of the scar.The levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were determined by enzyme-linked immunosorbent assay(ELISA).The expression levels of type collagenⅠ(COLⅠ)andα-smooth muscle actin(α-SMA)were detected by real-time fluorescence quantitative PCR.Tissue morphology was observed by HE staining.Results Compared with the corresponding homotype control,the ADSCs surface antigens CD73(99.9%)and CD90(99.9%)were positive,while CD34(2.09%)and CD45(1.36%)were negative,indicating that adipogenic stem cells were successfully extracted.ELISA results showed that compared with the control group at the corresponding time points,the expression of IL-6 was significantly down-regulated and the expression of TNF-αwas inhibited at 1,3 and 5 weeks after ADSCs treatment,with statistical significance(P<0.05).The results of real-time fluorescence quantitative PCR showed that COLⅠandα-SMA expression were significantly down-regulated at 1,3 and 5 weeks after ADSCs treatment compared with the corresponding time points in the control group(P<0.05).HE staining results showed that the addition of ADSCs could significantly reduce the aggregation of inflammatory cells in the tissues,while the disorder of collagen arrangement was more serious in the control group.Conclusion ADSCs can inhibit hypertrophic scar by down regulating the expression of inflammatory factors and inhibiting the contents of COLⅠandα-SMA.
作者
杨玲玲
黄悦
王洪一
白泽明
韩子阳
刘芮嫡
陶凯
YANG Ling-ling;HUANG Yue;WANG Hong-yi;BAI Ze-ming;HAN Zi-yang;LIU Rui-di;TAO Kai(Department of Burns and Plastic Surgery,General Hospital of Northern Theater Command,Shenyang 110016,China)
出处
《临床军医杂志》
CAS
2022年第5期503-506,509,共5页
Clinical Journal of Medical Officers
基金
国家自然科学基金面上项目(81971845)
国家自然科学基金青年基金(82002047)
关键词
增生性瘢痕
脂肪源性干细胞
炎症
创面愈合
Hypertrophic scar
Adipose-derived stem cells
Inflammation
Wound healing
