HA1077对骨髓间质干细胞促愈作用的影响  

Effect of HA1077 on mesenchymal stem cells promoting wound healing

作　　者：白晓东[1] 刘贤华[2] 柳晓杰[1] 王佳哲[1] 付小兵[3] 

机构地区：[1]武警总医院烧伤整形科,北京100039 [2]武警总医院中心实验室,北京100039 [3]解放军总医院第一附属医院全军创面修复重点实验室

出　　处：《中华创伤杂志》2009年第7期663-667,共5页Chinese Journal of Trauma

摘　　要：目的探讨HA1077对骨髓间质干细胞（bone mesenchymal stemcells，BMSCs）促愈作用的影响及其机制。方法（1）体外实验：体外分离培养大鼠BMSCs，随机分为正常对照组、诱导组和HA1077组。对照组加入DMEM完全培养液；诱导组：70％DMEM完全培养液＋30％大鼠成纤维细胞上清液＋含8μg/L表皮细胞生长因子（EGF）的培养液；HA1077组：在诱导组培养液基础上加入终浓度为10μmol／L的HA1077。取培养7d的细胞，流式细胞检测CK19表达，细胞周期和核增殖抗原（PCNA）表达。（2）在体实验：按前述方法制备兔BMSCs。取新西兰大耳白兔8只，在背部制备6个直径为3cm的圆形皮肤缺损（每侧各3个），深至皮下。创面随机分为空白对照组（A组）：创面应用等渗盐水；EGF对照组（B组）：创面外用EGF，50 U／cm^2；BMSCs治疗组（C组）：除用EGF外，创面注射BMSCs细胞悬液1ml，2×10^6／ml；BMSCs＋HA1077治疗组（D组）：除应用c组措施外，创面注射0．5mlHA1077，浓度为20μmol／L。伤后3，7，14d计算愈合指数（WCD。结果（1）体外实验：诱导组CK19的阳性率为（11．13±2．14）％，HA1077组为（40．31±0．82）％，明显高于诱导组（P〈0．01）。HA1077组PCNA阳性率明显高于诱导组，细胞周期分析显示S期细胞数量最多。（2）在体实验：D组伤后14d的创面愈合指数为86．20±1．92，效果明显好于B组、C组和对照组。结论HA1077可以促进BMSCs细胞表达PCNA进入S期，进而分化表达CK19；在体表联合应用HA1077和BMSCs，可以明显促进创面愈合。Objective To observe the effect and mechanism of HA1077 on bone mesenchymal stem cells （BMSCs） promoting wound healing. Methods （ 1 ） In vitro study： rats＇ BMSCs were harvested and divided into three groups randomly, ie, control group, induction group and HAl077 group. In control group, culture medium was DMEM with 10% fetal bovine serum. In induction group, induction medium was DMEM which consisted of 10% FBS, 8μg/ml EGF and epithelium culture condition medium. In HA1077 group, culture condition medium was the induction medium plus 10 p.mol/L HA1077. At day 7 after replanting, CK19 expression, PCNA expression and cell cycle among three groups were clarified by flow cytometry. （2） In vivo study ： rabbit＇ s BMSCs were isolated. Six pieces of full skin loss wound were created on the back of a rabbit, with three pieces of wound on the half side of back. The wound was round and 3 cm in diameter. Eight New Zealand rabbits were randomly divided into four groups, ie, control group （ Group A）, EGF control group （ Group B） , BMSCs group （ Group C ） and BMSCs ＋ HA1077 group （Group D）. Wounds were treated by normal saline in Group A and by EGF （50 U/cm^2） in Group B. Besides EGF, 1 ml of BMSCs suspension （2 × 10^6/ml） was used for wound repair in Group C.0.5 ml HAl077 （201xmol/L）, BMSCs and EGF were used together for wound repair in Group D. At days 3, 7 and 14 after injury, wound closure index （WCI） was calculated. Results In vitro study：the positive rate of CK19 in the HA1077 group was （ 40. 31 ± 0. 82）% , higher than （11.13 ±2.14）% in the induction group （P 〈0.01）. The PCNA positive rate and the S phasecell quantity in HAl077 group were also higher than that in the induction group. In vivo study： WCI was （86.20 ± 1.92）% in Group D at day 14 after injury, obviously higher than that in other three groups. Conclusions HA1077 can promote BMSCs＇ entering into S phase, expressing PCNA and differentiating and expressing CK19. In vivo combination

关 键 词：间质干细胞 细胞分化 创面愈合 兔 HA1077 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]