优化常温机械灌注对大鼠心脏性死亡后保存供体肝移植物胆管上皮细胞的影响

Q4 Medicine

中华肝胆外科杂志 Pub Date : 2020-02-28 DOI:10.3760/CMA.J.ISSN.1007-8118.2020.02.012

Bin Hou, H. Cao, Liu Yang, D. Sun, Yuan Shi, Z. Shen, Hongli Song

{"title":"优化常温机械灌注对大鼠心脏性死亡后保存供体肝移植物胆管上皮细胞的影响","authors":"Bin Hou, H. Cao, Liu Yang, D. Sun, Yuan Shi, Z. Shen, Hongli Song","doi":"10.3760/CMA.J.ISSN.1007-8118.2020.02.012","DOIUrl":null,"url":null,"abstract":"Objective \nTo study the effect of bone marrow mesenchymal stem cells (BMMSCs) combined with normothermic mechanical perfusion (NMP) on biliary epithelial cells (BEC) after DCD donor liver transplantation in rats. \n \n \nMethods \nThe third generation of BMMSCs and the BMMSCs modified by Ad/HO-1 (Ad/HO-1/BMMSCs) were cultured, identified and expanded in vitro. To establish a stable NMP system device in vitro. The DCD liver transplantation models were constructed in rats after cardiac ischemia for 30 minutes, 220 SD recipient rats were randomly divided into sham operation group (S group, n=44) static cold storage (SCS group, n=44) group, and simple NMP group (P group, n=44), BMMSCs combined with NMP group (BP group, n=44) and BMMSCs modified by Ad/HO-1 combine with NMP group (HBP group, n=44), NMP group, BP group and HBP group were subjected to vitro perfusion for 4h. The group were taken at 0, 1, 7 and 14 days after transplantation and the relevant indicators were detected, n=6 in each group. The survival rate of the recipient rats, liver function and pathological changes of the bile duct were observed. The expression of cytokeratin 19 (CK19) protein in BEC was detected by immunohistochemistry and Western blot. Apoptotic biliary epithelial cells were detected by TUNEL staining and the expression of apoptosis-related protein caspase-3 was detected by immunohistochemistry. \n \n \nResults \nThe survival time of HBP group was significantly prolonged for (5.6±0.8) d in SCS group vs. (18.4±2.0) d in NMP group, (20.5±1.5) d in BP group, (82.5±3.2) d in HBP group, the differences were statistically significant (all P<0.05). Compared with other groups, the HBP group and the BP group were significantly improved in liver function and biliary pathology, and the expression of CK19 protein in BEC was significantly increased [(0.81±0.02) in S group vs. (0.35±0.03) in SCS group, (0.47±0.02) in NMP group, (0.63±0.02) in BP group, (0.77±0.01) in HBP group on postoperative day (POD) 14], the differences were statistically significant (all P<0.05). The number of apoptosis and the expression of apoptosis-related protein caspase-3 in HBP group were significantly decreased [(10.0±1.2) in S group vs. (57.3±5.5) in SCS group, (40.1±4.6) in NMP group, (32.0±2.2) in BP group, (13.7±3.1) in HBP group on POD 14], the difference was statistically significant (all P<0.05). Compared with the BP group, the protective effect of the HBP group was more obvious, and the difference was statistically significant (P<0.05). \n \n \nConclusion \nBy the method of the BMMSCs modified by Ad/HO-1 combined with NMP in vitro preservation of rat, DCD donor liver can significantly improve the effect of BEC on rats and the survival rate after liver transplantation. \n \n \nKey words: \nLiver transplantation; Normothermic mechanical perfusion; Mesenchymal stromal cells; Biliary epithelial cells; Apoptosis","PeriodicalId":10021,"journal":{"name":"中华肝胆外科杂志","volume":"26 1","pages":"139-144"},"PeriodicalIF":0.0000,"publicationDate":"2020-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The effect of optimized normothermic mechanical perfusion on biliary epithelial cells in liver graft by preserving the donation after cardiac death in rats\",\"authors\":\"Bin Hou, H. Cao, Liu Yang, D. Sun, Yuan Shi, Z. Shen, Hongli Song\",\"doi\":\"10.3760/CMA.J.ISSN.1007-8118.2020.02.012\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Objective \\nTo study the effect of bone marrow mesenchymal stem cells (BMMSCs) combined with normothermic mechanical perfusion (NMP) on biliary epithelial cells (BEC) after DCD donor liver transplantation in rats. \\n \\n \\nMethods \\nThe third generation of BMMSCs and the BMMSCs modified by Ad/HO-1 (Ad/HO-1/BMMSCs) were cultured, identified and expanded in vitro. To establish a stable NMP system device in vitro. The DCD liver transplantation models were constructed in rats after cardiac ischemia for 30 minutes, 220 SD recipient rats were randomly divided into sham operation group (S group, n=44) static cold storage (SCS group, n=44) group, and simple NMP group (P group, n=44), BMMSCs combined with NMP group (BP group, n=44) and BMMSCs modified by Ad/HO-1 combine with NMP group (HBP group, n=44), NMP group, BP group and HBP group were subjected to vitro perfusion for 4h. The group were taken at 0, 1, 7 and 14 days after transplantation and the relevant indicators were detected, n=6 in each group. The survival rate of the recipient rats, liver function and pathological changes of the bile duct were observed. The expression of cytokeratin 19 (CK19) protein in BEC was detected by immunohistochemistry and Western blot. Apoptotic biliary epithelial cells were detected by TUNEL staining and the expression of apoptosis-related protein caspase-3 was detected by immunohistochemistry. \\n \\n \\nResults \\nThe survival time of HBP group was significantly prolonged for (5.6±0.8) d in SCS group vs. (18.4±2.0) d in NMP group, (20.5±1.5) d in BP group, (82.5±3.2) d in HBP group, the differences were statistically significant (all P<0.05). Compared with other groups, the HBP group and the BP group were significantly improved in liver function and biliary pathology, and the expression of CK19 protein in BEC was significantly increased [(0.81±0.02) in S group vs. (0.35±0.03) in SCS group, (0.47±0.02) in NMP group, (0.63±0.02) in BP group, (0.77±0.01) in HBP group on postoperative day (POD) 14], the differences were statistically significant (all P<0.05). The number of apoptosis and the expression of apoptosis-related protein caspase-3 in HBP group were significantly decreased [(10.0±1.2) in S group vs. (57.3±5.5) in SCS group, (40.1±4.6) in NMP group, (32.0±2.2) in BP group, (13.7±3.1) in HBP group on POD 14], the difference was statistically significant (all P<0.05). Compared with the BP group, the protective effect of the HBP group was more obvious, and the difference was statistically significant (P<0.05). \\n \\n \\nConclusion \\nBy the method of the BMMSCs modified by Ad/HO-1 combined with NMP in vitro preservation of rat, DCD donor liver can significantly improve the effect of BEC on rats and the survival rate after liver transplantation. \\n \\n \\nKey words: \\nLiver transplantation; Normothermic mechanical perfusion; Mesenchymal stromal cells; Biliary epithelial cells; Apoptosis\",\"PeriodicalId\":10021,\"journal\":{\"name\":\"中华肝胆外科杂志\",\"volume\":\"26 1\",\"pages\":\"139-144\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-02-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华肝胆外科杂志\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3760/CMA.J.ISSN.1007-8118.2020.02.012\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华肝胆外科杂志","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3760/CMA.J.ISSN.1007-8118.2020.02.012","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Medicine","Score":null,"Total":0}

引用次数: 0

摘要

目的研究骨髓间充质干细胞（BMMSCs）联合常温机械灌注（NMP）对大鼠DCD供肝移植后胆管上皮细胞（BEC）的影响。方法体外培养、鉴定和扩增第三代骨髓间充质干细胞和经Ad/HO-1修饰的骨髓基质干细胞（Ad/HO-1/BMMSCs）。建立稳定的NMP体外系统装置。在大鼠心脏缺血30min后建立DCD肝移植模型，将220只SD受体大鼠随机分为假手术组（S组，n=44）、静态冷藏组（SCS组，n=4）、单纯NMP组（P组，n=44）、骨髓间充质干细胞与NMP联合组（BP组，n=40）和Ad/HO-1修饰的骨髓间充体与NMP结合组（HBP组，n=44），NMP组、BP组和HBP组分别进行体外灌注4h。在移植后0、1、7和14天取组，检测相关指标，每组n=6。观察受体大鼠的存活率、肝功能和胆管病理变化。用免疫组织化学和蛋白质印迹法检测细胞角蛋白19（CK19）在BEC中的表达。TUNEL染色检测凋亡的胆管上皮细胞，免疫组化检测凋亡相关蛋白胱天蛋白酶-3的表达。结果HBP组生存时间SCS组（5.6±0.8）d明显延长，NMP组（18.4±2.0）d，BP组（20.5±1.5）d，HBP组（82.5±3.2）d，差异有统计学意义（均P<0.05），CK19蛋白在BEC中的表达在术后第14天显著增加[S组（0.81±0.02）vs.SCS组（0.35±0.03），NMP组（0.47±0.02），BP组（0.63±0.02）和HBP组（0.77±0.01）]，HBP组的细胞凋亡数量和凋亡相关蛋白胱天蛋白酶-3的表达在POD 14上显著降低[S组为（10.0±1.2）vs.SCS组为（57.3±5.5），NMP组为（40.1±4.6），BP组为（32.0±2.2），差异有统计学意义（均P<0.05）。与BP组相比，HBP组的保护作用更为明显，差异有统计学显著性（P<0.05），DCD供肝可显著提高BEC对大鼠的作用及肝移植后的存活率。关键词：肝移植；正常机械灌注；间充质基质细胞；胆管上皮细胞；细胞凋亡

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

The effect of optimized normothermic mechanical perfusion on biliary epithelial cells in liver graft by preserving the donation after cardiac death in rats

Objective To study the effect of bone marrow mesenchymal stem cells (BMMSCs) combined with normothermic mechanical perfusion (NMP) on biliary epithelial cells (BEC) after DCD donor liver transplantation in rats. Methods The third generation of BMMSCs and the BMMSCs modified by Ad/HO-1 (Ad/HO-1/BMMSCs) were cultured, identified and expanded in vitro. To establish a stable NMP system device in vitro. The DCD liver transplantation models were constructed in rats after cardiac ischemia for 30 minutes, 220 SD recipient rats were randomly divided into sham operation group (S group, n=44) static cold storage (SCS group, n=44) group, and simple NMP group (P group, n=44), BMMSCs combined with NMP group (BP group, n=44) and BMMSCs modified by Ad/HO-1 combine with NMP group (HBP group, n=44), NMP group, BP group and HBP group were subjected to vitro perfusion for 4h. The group were taken at 0, 1, 7 and 14 days after transplantation and the relevant indicators were detected, n=6 in each group. The survival rate of the recipient rats, liver function and pathological changes of the bile duct were observed. The expression of cytokeratin 19 (CK19) protein in BEC was detected by immunohistochemistry and Western blot. Apoptotic biliary epithelial cells were detected by TUNEL staining and the expression of apoptosis-related protein caspase-3 was detected by immunohistochemistry. Results The survival time of HBP group was significantly prolonged for (5.6±0.8) d in SCS group vs. (18.4±2.0) d in NMP group, (20.5±1.5) d in BP group, (82.5±3.2) d in HBP group, the differences were statistically significant (all P<0.05). Compared with other groups, the HBP group and the BP group were significantly improved in liver function and biliary pathology, and the expression of CK19 protein in BEC was significantly increased [(0.81±0.02) in S group vs. (0.35±0.03) in SCS group, (0.47±0.02) in NMP group, (0.63±0.02) in BP group, (0.77±0.01) in HBP group on postoperative day (POD) 14], the differences were statistically significant (all P<0.05). The number of apoptosis and the expression of apoptosis-related protein caspase-3 in HBP group were significantly decreased [(10.0±1.2) in S group vs. (57.3±5.5) in SCS group, (40.1±4.6) in NMP group, (32.0±2.2) in BP group, (13.7±3.1) in HBP group on POD 14], the difference was statistically significant (all P<0.05). Compared with the BP group, the protective effect of the HBP group was more obvious, and the difference was statistically significant (P<0.05). Conclusion By the method of the BMMSCs modified by Ad/HO-1 combined with NMP in vitro preservation of rat, DCD donor liver can significantly improve the effect of BEC on rats and the survival rate after liver transplantation. Key words: Liver transplantation; Normothermic mechanical perfusion; Mesenchymal stromal cells; Biliary epithelial cells; Apoptosis

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

中华肝胆外科杂志 Medicine-Gastroenterology

CiteScore

0.20

自引率

0.00%

发文量

7101

期刊介绍： Chinese Journal of Hepatobiliary Surgery is an academic journal organized by the Chinese Medical Association and supervised by the China Association for Science and Technology, founded in 1995. The journal has the following columns: review, hot spotlight, academic thinking, thesis, experimental research, short thesis, case report, synthesis, etc. The journal has been recognized by Beida Journal (Chinese Journal of Humanities and Social Sciences). Chinese Journal of Hepatobiliary Surgery has been included in famous databases such as Peking University Journal (Chinese Journal of Humanities and Social Sciences), CSCD Source Journals of China Science Citation Database (with Extended Version) and so on, and it is one of the national key academic journals under the supervision of China Association for Science and Technology.