7胰岛素干预对糖尿病大鼠骨折愈合及骨痂内细胞因子的影响

中国组织工程研究第20卷第29期2016–07–08出版Chinese Journal of Tissue Engineering ResearchJuly 8,2016Vol.20,No.29·研究原著·www.CRTER .org周强，男，1984年生，江西省丰城市人，汉族，上海交通大学医学院在职硕士，主治医师，主要从事创伤与关节置换方面研究。

通讯作者：蒋雷生，博士，主任医师，博士生导师，上海交通大学医学院附属新华医院骨科，上海市200092中图分类号:R318文献标识码:A 文章编号:2095-4344(2016)29-04269-08稿件接受：2016-04-24胰岛素干预对糖尿病大鼠骨折愈合及骨痂内细胞因子的影响周强1，陆骅1，王占朝2，张豪杰2，蒋雷生1(1上海交通大学医学院附属新华医院骨科，上海市200092；2上海交通大学医学院附属新华医院崇明分院骨科，上海市202150)引用本文：周强，陆骅，王占朝，张豪杰，蒋雷生.胰岛素干预对糖尿病大鼠骨折愈合及骨痂内细胞因子的影响[J].中国组织工程研究，2016，20(29):4269-4276.DOI:10.3969/j.issn.2095-4344.2016.29.002ORCID:0000-0001-8517-6854(周强)文章快速阅读：文题释义：细胞因子：是指主要由免疫细胞分泌的、能调节细胞功能的小分子多肽。

在免疫应答过程中，细胞因子对于细胞间相互作用、细胞的生长和分化有重要调节作用。

由于基因工程、细胞工程研究的飞速发展，不仅克隆了早先发现的生物活性肽的cDNA ，而且发现了许多新的细胞因子，并对各种细胞因子产生来源、分了子结构和基因、相应的受体、生物学功能以及与临床的关系等进行了大量的研究，成为当今基础免疫学和临床免疫学研究中一个活跃的领域。

胰岛素：胰岛素是由胰脏内的胰岛β细胞受内源性或外源性物质刺激而分泌的一种蛋白质激素。

胰岛素作为降糖药物被广泛作用于临床，是机体内降低血糖的最关键激素，研究还发现它同时具备促进骨组织再生的特性。

摘要背景：糖尿病是一种以高血糖为特征表现的内分泌代谢紊乱，从而损伤身体多个系统的全身性疾病，并且糖尿病患者骨折后的愈合情况一般都不理想。

目的：动态检测胰岛素治疗前后，糖尿病大鼠在骨折愈合过程中细胞因子的变化趋势。

方法：SD 大鼠120只，随机取90只静脉注射5%四氧嘧啶溶液建立糖尿病大鼠模型；另取30只静脉注射等量的生理盐水做对照。

糖尿病大鼠模型随机分为胰岛素治疗组和糖尿病组。

将3组大鼠右胫骨中段标准横断骨折，胰岛素治疗组每日胰岛素皮下注射1次，糖尿病组及对照组给予等容量的生理盐水。

次日检测大鼠血糖情况；骨折后第1，4，8周拍X 射线片观察大鼠骨折的愈合情况；造模后第4，6，8周测定每组大鼠离断后右胫骨骨痂的生物力学强度；免疫组织化学和原位杂交技术检测骨痂内多种细胞因子的表达情况。

结果与结论：①X 射线片结果显示：糖尿病组骨痂形成时间和骨折愈合速度明显低于胰岛素治疗组和对照组，而后2组差异无显著意义；造模后4，6，8周糖尿病组骨折处骨痂的机械强度均明显低于胰岛素治疗组和对照组，而后2组差异无显著意义；②骨折后1-4周：骨形态发生蛋白2、碱性成纤维细胞生长因子、转化生长因子、血管内皮生长因子在骨痂中广泛表达，糖尿病组中的表达明显低于和滞于胰岛素治疗组和对照组，而后2组差异无显著性意义；③结果表明：糖尿病大鼠骨折愈合过程中较正动态检测胰岛素治疗前后糖尿病大鼠在骨折愈合过程中细胞因子的变化趋势建模成功74只结果：糖尿病状态下会引起机体内骨吸收增加，骨形成减少，从而影响骨代谢并推迟骨折的愈合；用胰岛素干预治疗后，骨折部位细胞因子的水平提高，促进成骨细胞增殖和骨折愈合。

90只建立大鼠糖尿病模型胰岛素治疗组37只糖尿病组37只对照组30只右胫骨横断骨折实验血糖检测相关细胞因子原位杂交检测相关细胞因子免疫组化检测生物力学强度检测SD 大鼠120只Zhou Qiang,Master, Attending physician,Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200092,ChinaCorresponding author: Jiang Lei-sheng,M.D., Chief physician,Doctoral supervisor,Department of Orthopedics,Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200092,ChinaAbstract 常大鼠，骨痂形成速度和生物力学强度降低，骨折断处组织细胞的生长因子表达水平明显滞后和下降，运用胰岛素干预治疗后可显著提高骨折部位细胞因子的水平，促进成骨细胞增殖和骨折愈合。

关键词：组织构建；骨组织工程；糖尿病；胰岛素；骨痂；骨折愈合；细胞因子主题词：糖尿病；胰岛素；骨折愈合；细胞因子；组织工程Insulin effects on fracture healing and cytokines in the osteotylus in experimental diabetic ratsZhou Qiang1,Lu Hua1,Wang Zhan-chao2,Zhang Hao-jie2,Jiang Lei-sheng1(1Department of Orthopedics, Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai200092,China; 2Department of Orthopedics,Chongming Branch,Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai202150,China)AbstractBACKGROUND:Fracture healing in diabetic patients is usually unsatisfactory because of hormones and metabolic disorder,and an eventual multiple organ dysfunction resulting from high blood glucose. OBJECTIVE:To dynamically observe the changes of cytokines during the fracture healing process in diabetic rats before and after insulin treatment.METHODS:A total of120Sprague-Dawley rats were included in this study.Of them,90rats intravenously injected with5%tetraoxypyrimidine to induce rat models of diabetes were randomized into insulin treatment and diabetes groups,respectively.The remaining30rats were intravenously injected with equal volume of saline and selected as control group.The next day,blood glucose was determined.Healing at1, 4,and8weeks after fracture were observed by the X-ray film.Biomechanical strength of the injured right tibia was measured at4,6,and8weeks after modeling.Cytokines in the osteotylus were determined by immunohistochemical staining and in situ hybridization technique.RESULTS AND CONCLUSION:The X-ray films showed that the speed of fracture healing in the diabetes group was slower than insulin treatment and control groups.Biomechanical strength of the osteotylus in the diabetes group was significantly decreased compared with the insulin treatment and control groups. However,there were no significant differences in above-mentioned parameters between the control and insulin treatment groups.Bone morphogenetic protein2,basic fibroblast growth factor,transforming growth factor-beta,and vascular endothelial growth factor were widely expressed in the osteotylus and their expressions in diabetes group were significantly lower and slower than those in the control and insulin treatment groups.There was no statistical difference between control and insulin treatment groups.These results indicate that osteotylus formation speed,biomechanical strength,and growth factor expressions at the fracture site in diabetes rats were decreased compared with normal rats.Insulin treatment can enhance cytokine levels at the fracture site,thereby promoting the osteoblast proliferation and fracture healing. Subject headings:Diabetes Mellitus;Insulin;Fracture Healing;Cytokines;Tissue EngineeringCite this article:Zhou Q,Lu H,Wang ZC,Zhang HJ,Jiang LS.Insulin effects on fracture healing and cytokines in the osteotylus in experimental diabetic rats.Zhongguo Zuzhi Gongcheng Yanjiu.2016;20(29):4269-4276.0引言Introduction糖尿病可引发人体多系统的慢性损伤并导致多种相关的骨关节疾病，愈合情况一般较不理想[1-2]。