自体富血小板凝胶联合脂肪干细胞促进糖尿病足大鼠创面修复的机制研究  

Study on Mechanism of Autologous Platelet-rich Plasma Gel Combined with Adipose-derived Stem Cells on Promoting Wound Repair of Rats with Diabetic Foot

作　　者：冯召岚[1] 盛健健 董爱武[1] 李江雄[1] 曹玲玲[1] FENG Zhaolan;SHENG Jianjian;DONG Aiwu;LI Jiangxiong;CAO Lingling(Jiujiang No.1 People’s Hospital,Jiangxi Province,Jiujiang 332000,China;不详)

机构地区：[1]江西省九江市第一人民医院,江西九江332000

出　　处：《中国医学创新》2022年第23期24-30,共7页Medical Innovation of China

基　　金：九江市科技计划项目(S2021ZDYFN090)。

摘　　要：目的:分析自体富血小板(PRP)凝胶联合脂肪干细胞(ADSCs)干预糖尿病足大鼠对磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路的影响。方法:取78只清洁级雄性SD大鼠,随机选取15只为正常组,其余63只采用高糖高脂饲料喂养联合链脲佐菌素(STZ)腹腔注射诱导糖尿病大鼠模型,并在每只实验大鼠后足左侧做了一个5 mm×5 mm正方形皮肤全层缺损创面。共60只大鼠建模成功,根据创面处理方式分为4组:模型对照组、PRP组、ADSCs组、PRP+ADSCs组各15只。分别于创伤后3、7、10、14 d时,每组随机取3只大鼠,观察创面愈合情况,通过免疫组化法比较各组创面组织血小板-内皮细胞黏附分子(CD31)阳性细胞分布情况及PI3K、Akt蛋白表达情况;采用实时荧光定量PCR(RT-PCT)检测各组干预3、7、10、14 d时PI3K、Akt mRNA相对表达水平。结果:PRP+ADSCs组、正常组、ADSCs组、PRP组大鼠创面完全愈合时间均显著早于模型对照组(P<0.05),且PRP+ADSCs组与正常组的完全愈合时间最早,而两组间比较差异无统计学意义(P>0.05),各组大鼠创口愈合率的组间及时间点比较差异有统计学意义(P<0.05);免疫组化结果显示,干预3、7、10 d时PRP+ADSCs组、正常组、ADSCs组、PRP组的CD31细胞阳性率及PI3K、Akt表达OD值均显著高于模型对照组(P<0.05),且干预3、7、10 d时PRP+ADSCs组与正常组的上述指标显著高于其他组(P<0.05);RT-PCR分析显示,当将模型对照组设为单位一时,PRP+ADSCs组、正常组、ADSCs组、PRP组大鼠在创面干预3、7、10 d时,分别出现不同程度的PI3K、Akt mRNA表达上调。结论:自体PRP凝胶联合ADSCs干预可促进糖尿病足大鼠新生血管生成及创口愈合,这可能与PI3K/Akt信号通路的激活相关。Objective:To analyze the effects of autologous platelet-rich plasma(PRP)gel combined with adipose-derived stem cells(ADSCs)on phosphoinositide-3-kinase(PI3K)/protein kinase B(Akt)signaling pathways in rats with diabetic foot.Method:A total of 78 clean male SD rats were taken,among which 15 rats were randomly selected as normal group and the remaining 63 rats were fed with high-glucose and high-fat diet combined with intraperitoneal injection of Streptozotocin(STZ)to induce diabetic rat models.A 5 mm×5 mm square full-thickness skin defect wound was made on the left side of the posterior foot of each experimental rat.A total of 60 rats were successfully modeled,and they were divided into four groups according to the wound treatment method,including model control group,PRP group,ADSCs group and PRP+ADSCs group,with 15 rats in each group.At 3,7,10 and 14 d after trauma,3 rats were randomly selected from each group to observe the wound healing,and the distribution of platelet-endothelial cell adhesion molecule(CD31)positive cells and the protein expressions of PI3K and Akt in wound tissues of each group were compared by immunohistochemistry.Real-time fluorescent quantitative PCR(RT-PCT)was used to detect the relative mRNA expression levels of PI3K and Akt in each group at 3,7,10 d and 14 d of intervention.Result:The wound complete healing time of rats in PRP+ADSCs group,normal group,ADSCs group and PRP group were significantly earlier than that in model control group(P<0.05),and the PRP+ADSCs group and the normal group had the shortest complete healing time,the difference between groups was not statistically significant(P>0.05),and there was a statistically significant difference in the wound healing rate between the groups and at different time points in each group(P<0.05).Immunohistochemistry showed that the positive rate of CD31 cells and the expressions of PI3K and Akt in PRP+ADSCs group,normal group,ADSCs group and PRP group were significantly higher than those in model control group at 3,7 and 10 d of intervent

关 键 词：PI3K/AKT信号通路 糖尿病足 大鼠 免疫组化 实时荧光定量PCR 

分 类 号：R587.2[医药卫生—内分泌]