脐带血富血小板血浆促进糖尿病大鼠难愈合创口修复作用研究  

Experimental study on effects of cord blood PRP promoting repairs of refractory wounds in diabetic rats

作　　者：杜为 张莉 张凯悦 赵晶磊 李宗金[1] 韩俊领 DU Wei;ZHANG Li;ZHANG Kaiyue;ZHAO Jinglei;LI Zongjin;HAN Junling(School of Medicine,Nankai University,Tianjin 300071,China;Union Stem Cell&Gene Engineering Co.,LTD,Tianjin 300384,China;Tianjin Enterprises Key Laboratory of Blood Cell Therapy Technology,Tianjin 300384,China;National Stem Cell Product Industrialization Base,Tianjin 300384,China)

机构地区：[1]南开大学医学院,天津300071 [2]协和干细胞基因工程有限公司,天津300384 [3]天津市血液细胞治疗技术企业重点实验室,天津300384 [4]国家干细胞产品产业化基地,天津300384

出　　处：《药物评价研究》2021年第4期715-720,共6页Drug Evaluation Research

基　　金：天津市科技计划创新平台专项(18PTSYJC00070);天津市博士后择优资助计划项目(TJQYBSH2018030)。

摘　　要：目的探讨脐带血富血小板血浆(CB-PRP)对糖尿病性难愈合创口修复的促进作用。方法ELISA试剂盒法检测CB-PRP和外周血PRP(PB-PRP)的血管内皮生长因子(VEGF)和血小板衍生生长因子BB(PDGF-BB)的含量;SD大鼠随机分为对照组、模型组、PB-PRP组和CB-PRP组,除对照组外,按65 mg/kg剂量1次性ip链脲佐菌素(STZ)溶液诱发糖尿病模型;所有大鼠均建立全层皮肤缺损创伤模型。测定创伤再上皮化速率;术后10 d,采集组织学标本,进行苏木精伊红(HE)染色、天狼星红染色和CD31免疫荧光染色。结果CB-PRP中VEGF、PDGF-BB含量高于PB-PRP,其中VEGF差异显著(P<0.05)。术后10、14、21 d,PB-PRP和CB-PRP组创口再上皮化速率显著快于模型组(P<0.05),术后21 d时,CB-PRP组的再上皮化率(92.8%)显著快于PB-PRP组(84.6%)(P<0.05)。HE染色结果显示,与模型组比较,PB-PRP和CB-PRP组肉芽组织生长更为旺盛,可见胶原纤维存在较多,新生的毛细血管较多。天狼猩红染色结果显示,PB-PRP和CB-PRP组创面增生区胶原纤维丰富,以Ⅰ型胶原为主,其间见Ⅲ型胶原。CD31免疫荧光染色结果显示,与模型组比较,CB-PRP和PB-PRP组毛细血管数量显著增加(P<0.05);与PB-PRP组比较,CB-PRP组的毛细血管数量显著增加(P<0.05)。结论CB-PRP具有促进糖尿病性难愈合皮肤创口修复的作用,有望成为组织工程治疗的新方法。Objective To investigate the promoting effect of cord blood-derived PRP(CB-PRP)on the diabetic refractory wound repair.Methods We detected the content of Vascular Endothelial Growth Factor(VEGF)and Platelet-Derived Growth Factor BB(PDGF-BB)in CB-PRP and peripheral blood-derived PRP(PB-PRP)by ELISA kit method.SD rats were randomly divided into control group,model group,PB-PRP group and CB-PRP group.Except for the control group,diabetic model was induced by once ip streptozotocin(STZ)solution at a dose of 65 mg/kg.A full-thickness skin defect model was established with all rats.The effects of refractory skin wound healing were compared including rate of re-epithelialization and histological examination.Hematoxylin eosin(HE)staining,Sirius red staining and CD31 immunofluorescence staining were performed respectively to detect the wound healing process in histology.Results The level of VEGF in CB-PRP group was significantly higher than that of PB-PRP(P<0.05).CBPRP group showed higher level of PDGF-BB than PB-PRP,but there was no statistically significant difference between two groups.The re-epithelization rate of PB-PRP and CB-PRP group was significantly faster than that of model group at 10,14 and 21 days after operation(P<0.05).At 21 days after operation,the re-epithelization rate of CB-PRP group(92.8%)was significantly faster than that of PB-PRP group(84.6%)(P<0.05).HE staining results showed that compared with model group,the granulation tissue of PB-PRP and CB-PRP groups grew more vigorously,and there were more collagen fibers and new capillaries.The results of Sirius red staining showed that there were abundant collagen fibers in the proliferative area of the wounds in PB-PRP and CB-PRP groups,mainly type Ⅰ collagen,with type Ⅲ collagen in between.CD31 immunofluorescence staining results showed that,compared with model group,the number of capillaries in CB-PRP and PB-PRP groups was significantly increased(P<0.05);Compared with PBPRP group,the number of capillaries in CB-PRP group was significantly increased

关 键 词：脐带血 富血小板血浆 生长因子 糖尿病 难愈合创口 修复 再上皮化 

分 类 号：R965[医药卫生—药理学]