木葡聚糖基温敏水凝胶预防大鼠腹壁-肠壁再粘连及其对局部TGF-β1和CTGF表达的影响

doi:10.13481/j.1671-587x.20180204

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (02): 223-228.doi: 10.13481/j.1671-587x.20180204

木葡聚糖基温敏水凝胶预防大鼠腹壁-肠壁再粘连及其对局部TGF-β1和CTGF表达的影响

孙红¹, 李硕峰¹, 崔菁¹, 张二帅², 李景武³, 熊艳杰⁴, 马丽桃¹, 姚芳莲², 车鹏程¹

1. 华北理工大学基础医学院河北省慢性病重点实验室唐山市慢性病临床基础研究重点实验室, 河北唐山 063210;
2. 天津大学化工学院高分子科学与工程系, 天津 300350;
3. 河北省唐山市人民医院腹盆外科, 河北唐山 063001;
4. 华北理工大学附属医院病理科, 河北唐山 063000

收稿日期:2017-06-30 出版日期:2018-03-28 发布日期:2018-03-30
通讯作者: 车鹏程,教授,硕士研究生导师(Tel:0315-8805157,E-mail:xsun1@hotmail.com) E-mail:xsun1@hotmail.com
作者简介:孙红(1972-),女,河北省唐山市人,教授,医学博士,主要从事组织工程和再生医学方面的研究。
基金资助:
国家自然科学基金资助课题（81101448）；河北省科技厅自然科学基金资助课题（H2017209005）；河北省人事厅引进留学人员资助项目资助课题（C201400560）；河北省唐山市科技创新团队培养计划资助课题（冀唐社函[2017]12号）

Preventive effect of MXG hycrogel on recurrentadhesion between abdominal wall and cecum and its influence in expressions of TGF-β1 and CTGF in rats

SUN Hong¹, LI Shuofeng¹, CUI Jing¹, ZHANG Ershuai², LI Jingwu³, XIONG Yanjie⁴, MA Litao¹, YAO Fanglian², CHE Pengcheng¹

1. Key Laboratory for Chronic Diseases of Hebei Province, Key Laboratory for Preclinical and Basic Research on Chronic Diseases of Tangshan City, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063210, China;
2. Department of Polymer Science and Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China;
3. Department ofAbdominal and Pelvic Surgery, Tangshan People's Hospital, Tangshan 063000, China;
4. Department of Pathology, Affiliated Hospital, North China University of Science and Technology, Tangshan 063000, China

Received:2017-06-30 Online:2018-03-28 Published:2018-03-30

摘要/Abstract

摘要： 目的：评价木葡聚糖基（mXG）温敏水凝胶对L929鼠成纤维细胞的抗黏附性能，建立粘连松解术后再次粘连动物模型，探讨mXG温敏水凝胶对粘连松解术后再次粘连的预防作用及对转化生长因子β1（TGF-β1）和结缔组织生长因子（CTGF）表达的影响。方法：将L929鼠成纤维细胞接种于mXG温敏水凝胶表面，荧光显色检测其在凝胶表面的黏附性能。制备粘连松解术后再次粘连大鼠模型。48只SD大鼠二次损伤完成后随机分成3组，分别为再粘连组，将1 mL生理盐水注射于大鼠腹壁和盲肠的创面；商业膜组，将2 cm×3 cm大小的壳聚糖防粘连膜覆盖于腹壁创面；mXG温敏水凝胶组，将1mL4% mXG温敏水凝胶注射至腹壁和盲肠的创面，待水凝胶完全形成（3 min）后关腹；每组16只。二次术后第7和14天，每组分别处死8只大鼠，对粘连程度进行评分及组织学观察，免疫组织化学法观察TGF-β1和CTGF表达水平。结果：对照组可见大量L929鼠成纤维细胞贴壁生长，呈细长的梭形并有伪足的形成；mXG温敏水凝胶组在凝胶表面仅有非常少的L929鼠成纤维细胞，呈圆球状。一次术后所有大鼠均形成了可用钝器分开的粘连区。二次术后第7天，再粘连组形成了4~5分粘连，纤维结缔组织大量增生；商业膜组大部分为中度粘连，纤维结缔组织增生较多；mXG组大部分大鼠未见粘连发生且盲肠及腹壁开始愈合，有少量纤维结缔组织增生。术后第14天，再粘连组大鼠腹腔粘连严重，纤维结缔组织及胶原大量增生；商业膜组形成了接近再粘连组的严重粘连；mXG组有2只大鼠发生轻微粘连，其余均未发生粘连，并且盲肠及腹壁创面恢复良好。术后第7和14天mXG水凝胶组大鼠平均粘连评分明显低于再粘连组和商业膜组（P<0.05）。免疫组织化学染色，TGF-β1和CTGF表达水平随大鼠腹膜粘连评分和胶原沉积升高而升高，在再粘连组表达最高，在mXG温敏水凝胶组表达最低。结论：mXG温敏水凝胶具有优异的抗成纤维细胞黏附性能，可以有效预防粘连松解术后再次粘连的发生，并可减少粘连相关因子TGF-β1和CTGF的表达水平。

关键词: 粘连松解术, 转化生长因子β1, 木葡聚糖基温敏水凝胶, 再次粘连, 结缔组织生长因子

Abstract: Objective:To evaluate the anti-adhesion properties of xyloglucan(mXG)hydrogel in the L929 mouse fibroblasts,to establish the animal model of recurrent adhesion in the rats after adhesionlysis,and to investigate the prevention effect of mXG hydrogel in recurrent adhesion and its influence in the expressions of transforming growth factor-β1(TGF-β1) and connective tissue growth factor (CTGF). Methods: After seeding on the surface of mXG gel,the adhesion property of L929 mouse fibroblasts was detected with fluorescence staining.The rat models of recurrent adhesion were established after adhesionlysis.Fourty-eight SD rats were randomly divided into 3 groups respectively(n=16).In adhesion group,1 mL saline was injected into the abdominal wall and cecum of the rats; in commercial membrane group,the 2 cm×3 cm chitosan anti-adhesion membrane was used to cover the wound of abdominal wall of the rats;in mXG hydrogel group,1 mL 4% mXG hydrogel was injected into the abdominal wall and cecal wound of the rats,and abdominal surgery was ended after the complete formation of the hydrogel (3 min).Eight rats were killed in each group 7 and 14 d after the second operation,and the degrees of adhesion were evaluated and the histological changes were observed.Immunohistochemical staining was used to observe the expression levels of CTGF and TGF-β1. Results: A large number of L929 mouse fibroblasts proliferated well and exhibited a spherical morphology in control group; but in mXG hydrogel group,only very little L929 mouse fibroblasts were globular,showing the mXG hydrogel had good resistance to the adhesion of L929 mouse fibroblasts.Blunt separate adhesion surface formed in all of the rats after operation.7 d after the second operation, 4-5 score adhesion with fibrous connective tissue hyperplasia formed in adhesion group;moderate adhesion formed in commercial membrane group with more connective tissue;most cecum and abdominal wall began to heal in mXG hydrogel group with less connective tissue.14 d after the second operation,more severe peritoneal adhesions presented in adhesion group with proliferated fiber connetive tissue and collegan;severe adhesions also presented in commercial membrane group; mild adhesion was found in two rats mXG group,the other rats had no adhesion, and the defects were almost completely recovered.On days 7 and 14 after the second operation,the mean adhesion score of the rats in mXG group was significantly lower than those in adhesion group and commercial membrane group(P<0.05).The expression levels of TGF-β1 and CTGF were increased with the increase of peritoneal adhesion scores and collagen deposition;the expression levels of TGF-β1 and CTGF were the highest in adgesion group and the lowest in mXG group. Conclusion: mXG hydrogels have good resistance to fibroblast adhesion,and mXG hydrogel is effective in reducing the formation of recurrent adhesion in the rats after adhesionlysis and can decrease the expression levels of adhesion-related factors TGF-β1 and CTGF.

Key words: xyloglucan hydrogel, transforming growth factor β1, recurrent adhesion, adhesionlysis, connective tissue growth factor

中图分类号:

R656

孙红, 李硕峰, 崔菁, 张二帅, 李景武, 熊艳杰, 马丽桃, 姚芳莲, 车鹏程. 木葡聚糖基温敏水凝胶预防大鼠腹壁-肠壁再粘连及其对局部TGF-β1和CTGF表达的影响[J]. 吉林大学学报(医学版), 2018, 44(02): 223-228.

SUN Hong, LI Shuofeng, CUI Jing, ZHANG Ershuai, LI Jingwu, XIONG Yanjie, MA Litao, YAO Fanglian, CHE Pengcheng. Preventive effect of MXG hycrogel on recurrentadhesion between abdominal wall and cecum and its influence in expressions of TGF-β1 and CTGF in rats[J]. Journal of Jilin University Medicine Edition, 2018, 44(02): 223-228.

参考文献

[1] Diamond MP,Wexner SD,diZereg GS,et al.Adhesion prevention and reduction:current status and future recommendations of a multinational interdisciplinary consensus conference[J].Surg Innov,2010,17(3):183-188.
[2] Brochhausen C,Schmitt VH,Planck CN,et al.Current strategies and future perspectives for intraperitoneal adhesion prevention[J].J Gastrointest Surg,2012,16(6):1256-1274.
[3] Ward BC,Panitch A.Abdominal adhesions:current and novel therapies[J].J Surg Res,2011,165:91-111.
[4] 刘国辉,李有柱,赵军.羧甲基纤维素钠在预防术后腹膜粘连中的作用[J].吉林大学学报:医学版,2002,28(2):163-164.
[5] Yang B,Gong C,Zhao X,et al.Preventing postoperative abdominal adhesions in a rat model with PEG-PCL-PEG hydrogel[J].Int J Nanomed,2012,7:547-557.
[6] Gao X,Deng X,Wei X,et al.Novel thermosensitive hydrogel for preventingformation of abdominal adhesions[J].Int J Nanomed,2013,8:2453-2463.
[7] Wu Q,Wang N,He T,et al.Thermosensitive hydrogel containing dexamethasone micelles for preventing postsurgical adhesion in a repeated-injury model[J].Sci Rep,2015,5:13553.
[8] Zhang E,Li J,Zhou Y,et al.Biodegradable and injectable thermoreversible xyloglucan based hydrogel forprevention of postoperative adhesion[J].Acta Biomater,2017,55:420-433.
[9] Nair SK,Bhat IK,Aurora AL.Role of proteolytic enzyme in the prevention of postoperative intraperitoneal adhesions[J].Arch Surg,1974,108(6):849-853.
[10] Okabayashi K,Ashrafian H,Zacharakis E,et al.Adhesions after abdominal surgery:a systematic review of the incidence,distribution and severity[J].Surg Today,2014,44(3):405-420.
[11] Yang Y,Liu X,Li Y,et al.A postoperative anti-adhesion barrier based on photoinduced imine-crosslinking hydrogel withtissue-adhesive ability[J].Acta Biomater,2017,62:199-209.
[12] Chen CH,Chen SH,Mao SH,et al.Injectable thermosensitive hydrogel containing hyaluronic acid and chitosan as a barrier for prevention of postoperative peritoneal adhesion[J].Carbohydr Polym,2017,173:721-731.
[13] Bae SH,Son SR,Kumar Sakar S,et al.Evaluation of the potential anti-adhesion effect of the PVA/Gelatin membrane[J].J Biomed MaterResPart B Appl Biomater,2014,102(4):840-849.
[14] Wei G,Chen X,Wang G,et al.Effect of resveratrol on the prevention of intra-abdominal adhesion formation in a rat model[J].Cell Physiol Biochem,2016,39(1):33-46.
[15] Zhang H,Song Y,Li Z,et al.Evaluation of ligustrazine on the prevention of experimentally induced abdominal adhesions in rats[J].Int J Surg,2015,211(6):115-121.
[16] Zhang H,Song Y,Li Z,et al.Evaluation of breviscapine on prevention of experimentally induced abdominal adhesions in rats[J].Am J Surg,2016,211(6):1143-1152.
[17] Yigitler C,Karakas DO,Kucukodaci Z,et al.Adhesion-preventing properties of 4% icodextrin and canola oil:a comparative experimental study[J].Clinics,2012,67(11):1303-1308.
[18] Thaler K,Mack JA,Berho M,et al.Coincidence of connective tissue growth factor expression with fibrosis and angiogenesis in postoperative peritoneal adhesion formation[J].Eur Surg Res,2005,37(4):235-241.
[19] Ryan CK,Sax HC.Evaluation of a carboxymethylcellulose sponge for prevention of postoperative adhesions[J].Am J Surg,1995,169(1):154-159.

木葡聚糖基温敏水凝胶预防大鼠腹壁-肠壁再粘连及其对局部TGF-β1和CTGF表达的影响

Preventive effect of MXG hycrogel on recurrentadhesion between abdominal wall and cecum and its influence in expressions of TGF-β1 and CTGF in rats

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