胰岛素样生长因子I和单纯疱疹病毒胸苷激酶基因共转染优化创面愈合的研究  

Study on cotransfection of genes of insulin-like growth factor I and herpes simplex virus thymidine kinase for optimization of wound healing

作　　者：杨磊[1] 王甲汉[1] 高建华[2] 

机构地区：[1]南方医科大学南方医院烧伤科,广州510515 [2]南方医科大学南方医院整形科,广州510515

出　　处：《中华烧伤杂志》2010年第3期202-206,共5页Chinese Journal of Burns

基　　金：广东省自然科学基金（9151051501000067）;南方医院院长基金（20088024）

摘　　要：目的 了解胰岛素样生长因子I（IGF-I）和单纯疱疹病毒胸苷激酶（HSV-tk）基因共转染对创面愈合的影响.方法 将30只雄性Wistar大鼠造成30%TBSAⅢ度烫伤,按随机数字表法分为A组（注射4.6 Pμg pcDNA3.1/IGF-I＋脂质体2000＋生理盐水）;B组（注射3.6μg pcDNA3.1/HSV-tk＋脂质体2000＋生理盐水）;C1组和c2组（均注射2.3μg pcDNA3.1/IGF.I＋1.8 μg pcDNA3.1/HSV-tk＋脂质体2000＋生理盐水）;D组（注射3.0 μg pcDNA3.1＋脂质体2000＋生理盐水）.每组6只大鼠,均于伤后即刻及7、14、21、28 d于大鼠左后背部创缘皮下注射上述混合物,其中C2组大鼠另于伤后29、30、31、32 d皮下注射丙氧鸟苷（2.5 mg/100 g）.称量烫伤大鼠体质量,计算创面愈合率.伤后35 d,免疫组织化学染色检测IGF-I基因在创面局部及肝脏组织的表达,放射免疫方法检测血清中IGF-I水平,RT-PCR检测HSV-tk基因在创面局部的表达,透射电镜观察C1、C2组Fb凋亡情况.数据采用单因素方差分析、Turkey法处理.结果 A、C1和C2组大鼠体质量于伤后7～35 d呈现增加趋势,与B、D组比较,差异具有统计学意义（F值分别为2.764、4.519、5.009、13.449、5.877,P值均小于0.05）;该3组大鼠创面愈合率高于B、D组（F值分别为5.286、100.880、152.380、127.850、147.750,P值均小于0.05）.A、C1和C2组大鼠创面组织Fb中IGF-I出现阳性表达,各组大鼠肝脏组织中未出现IGF-I阳性表达.各组大鼠血清中IGF-I含量为（1185±170）～（1270±130）ng/mL,差异无统计学意义（F=0.355,P=0.838）.B、C1、C2组大鼠创面组织中HSV-tk基因呈阳性表达.透射电镜显示C2组大鼠Fb出现凋亡,C1组大鼠未见此现象.结论 利用脂质体将pcDNA3.1/IGF-I peDNA3.1/HSV-tk基因转染于烫伤大鼠创面周围,可促进创面愈合,对瘢痕增生有一定抑制作用.Objective To study the effect of cotransfection of genes of insulin-like growth factor I （IGF-I） and herpes simplex virus thymidine kinase （HSV-tk） on wound healing. Methods Thirty male Wistar rats were inflicted with 30% TBSA full-thickness scald. They were then divided into A group （4. 6 μg pcDNA3.1/IGF- I ＋ Lipofectmine 2000 ＋ saline） , B group （3.6 μg pcDNA3. 1/HSV-tk ＋ Lipo-fectmine 2000 ＋ saline） , C1 group and C2 group （2. 3 μg pcDNA3.1/IGF-I ＋ 1. 8 μg pcDNA3. 1/HSV-tk ＋ Lipofectmine 2000 ＋ saline） , and D group （3.0 μg pcDNA3. 1 ＋ Lipofectmine 2000 ＋ saline） according to the random number table, with 6 rats in each group. The above-mentioned mixtures were subcutaneously injected into left back of each rat the moment after injury and on post scald day （PSD） 7, 14, 21 , and 28. Gancyclovir （2.5 mg/100 g） was hypodermically injected into rats in C2 group on PSD 29, 30, 31, 32. Changes in body weight of rats were measured. Wound healing rates were calculated. On PSD 35, the expressions of IGF- I gene in local wound and liver tissue were determined with immunohistochemical staining. The serum expression of IGF- I was determined with radioimmunoassay. Expression of HSV-tk gene in local wound was determined with RT-PCR. Apoptosis of fibroblast in C1 and C2 groups was observed under transmission electron microscope. Data were processed with one-way analysis of variance and Turkey method. Results Body weight of rats in A, C1, and C2 groups increased from PSD 7 through 35, and the difference between former three groups and B, D groups was statistically significant （with F value respectively 2.764, 4.519, 5.009, 13.449, 5.877, P values all below 0.05）. Wound healing rates of rats in A, C1, and C2 groups were higher than those in B, D groups （with F value respectively 5.286, 100. 880, 152.380, 127.850, 147. 750, P values all below 0.05）. IGF-I gene was positively expressed in wound fi-broblast in A, C1 and C2 groups, but negatively in liver tissues of all

关 键 词：烧伤 胰岛素样生长因子I 单纯疱疹病毒属 创面愈合 基因治疗 

分 类 号：R[医药卫生]