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}
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
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.
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