吉林大学学报（医学版）2026，Vol.52Issue(2)：318-329,12.DOI:10.13481/j.1671-587X.20260203
局部应用负载外泌体的DCC水凝胶对小鼠腹腔粘连的改善作用及其机制
Improvement effect of topical application of exosome loaded hydrogel DCC on peritoneal adhesion in mice and its mechanism
摘要
Abstract
Objective:To investigate the ameliorative effect of the injectable oxidized dextran(ODex)/carboxymethyl chitosan(CMCS)hydrogel(DCC)loaded with exosomes(Exo)(DCC/Exo)on postoperative peritoneal adhesion(PA)in the mice,and to elucidate its mechanism.Methods:The exosomes were extracted from the rats,and DCC/Exo was prepared and its chemical structure was characterized.Immunohistochemical staining method was used to detect the expression of the inflammatory factor Ly-6G in mouse dorsal skin tissue after injection of DCC/Exo to evaluate its biocompatibility.The mouse PA model was established by cecal abrasion.The KM mice were randomly divided into control group,hyaluronic acid(HA)group,DCC group,and DCC/Exo group.The PA condition was observed and scored in the mice in various groups at 5 and 14 d after operation.HE staining was used to observe the pathomorphology of PA tissue of the mice in various groups.At 5 d after operation,immunohistochemical staining method was used to detect the expression levels of myeloperoxidase(MPO)in PA tissue of the mice in various groups.The dihydroethidium(DHE)fluorescence probe assay was used to detect the levels of reactive oxygen species(ROS)in PA tissue of the mice in various groups;immunofluorescence staining was used to detect the expression levels of the oxidative damage marker 8-hydroxy-2'-deoxyguanosine(8-OHdG)and the mesothelial-mesenchymal transition(MMT)-related proteins E-cadherin and Vimentin in PA tissue of the mice in various groups.The Mitotracker fluorescence probe assay was used to detect the degree of mitochondrial damage in PA tissues of the mice in various groups.Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining was used to detect the apoptotic rates of the cells in PA tissue of the mice in various groups.Results:The isolated Exos exhibited a cup-shaped double-membrane structure.After loading with Exos,the characteristic absorption peaks of the DCC hydrogel showed no significant shift.The immunohistochemistry analysis results revealed only minimal neutrophil infiltration in mouse dorsal skin tissues.At 5 and 14 d after operation,compared with control group and DCC group,the mean adhesion score of the mice in DCC/Exo group was significantly decreased(P<0.001).The HE staining results showed severe PA with massive inflammatory cell infiltration of the mice in control group,mitigated PA in the mice in HA group and DCC group,and no adhesion with only a few inflammatory cells observed in the abdominal cavity of mice in DCC/Exo group.At 5 d after operation,the immunohistochemical staining results showed significant diffuse inflammatory infiltration in PA tissue of the mice in control group.Compared with control group and DCC group,the expression level of MPO in PA tissue of the mice in DCC/Exo group was significantly decreased(P<0.001).The DHE staining results showed that compared with control group and DCC group,the ROS level in PA tissue of the mice in DCC/Exo group was significantly decreased(P<0.001).The immunofluorescence staining results showed that compared with control group and DCC group,the expression levels of 8-OHdG and Vimentin in PA tissue of the mice in DCC/Exo group were significantly decreased(P<0.001),while the expression level of E-cadherin was significantly increased(P<0.001).Compared with control group and DCC group,the fraction of Mitotracker fluorescence area in PA tissue of the mice in DCC/Exo group was significantly increased(P<0.001),indicating a decreased degree of mitochondrial damage.Compared with control group and DCC group,the apoptotic rate of the cells in PA tissue of the mice in DCC/Exo group was significantly decreased(P<0.001).Conclusion:Topical application of the DCC/Exo composite hydrogel may effectively attenuate postoperative PA in the mice,and its mechanism may be related to the suppression of local inflammatory infiltration,reduction of tissue oxidative stress,decrease in apoptosis,and inhibition of the MMT process.关键词
术后腹腔粘连/水凝胶/外泌体/氧化应激/间皮-间质转化Key words
Postoperative peritoneal adhesion/Hydrogel/Exosome/Oxidative stress/Mesothelial-mesenchymal transition分类
医药卫生引用本文复制引用
马宁,孙红,柳爽,王成瑶,卢嘉骏,陈琳渝,王梓祎,赵伟钦,王伟潼,车鹏程..局部应用负载外泌体的DCC水凝胶对小鼠腹腔粘连的改善作用及其机制[J].吉林大学学报（医学版）,2026,52(2):318-329,12.基金项目
河北省科技厅自然科学基金项目(H2023209021,H2024209013) （H2023209021,H2024209013）
河北省教育厅大学生创新创业训练计划项目(202410081057) （202410081057）
