The Quantitative Research in Hepatocyte Apoptosis Level from Rabbit Donors after Brain Death_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 ZHONGZi-biao ¹ , ZHANGMeng ¹ , XIONGYan ¹ , WANGYan-feng ¹ ,  YEQi-fa ^1,2

1. Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan 430071, Hubei Province, China;
2. The 3rd Xiangya Hospital of Central South University, Research Center of National Health Ministry on Transplantation Medicine Engineering and Technology, Changsha 410013, Hunan Province, China;

Corresponding author：

YEQi-fa, Email: yqf_china@163.com

Keywords：

Brain death; Liver; Cell apoptosis

DOI：

10.7507/1007-9424.20160006

Video：

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Objective After establishing the rabbit brain death model, TUNEL, western blotting, and immuno-histochemical methods were used to detect hepatocyte apoptosis to study hepatocyte apoptosis level from rabbit donors after brain death. Methods Sixty healthy male New Zealand rabbits were divided into brain death group (n=30) and sham group (n=30). The rabbits of brain death group were established by increasing intracranial pressure in a modified, slow, and intermittent way, collecting liver tissues after corresponding treatment respectively, using TUNEL to detect apoptosis rate, western blotting and immunohistochemical methods to detect the expression of Cleaved-caspase 3. Results The hepatocyte apoptosis rate at each time point of brain death group were higher than those of the corresponding time point of sham group (P<0.05), and the rate of hepatocyte apoptosis increased gradually with the extension of brain death time (P<0.05). The results of Western blot assay and immunohistochemistry assay showed that the relative expression amount of Cleaved-caspase 3 protein increased gradually with the extension of brain death time (P<0.05), and relative expression amount of Cleaved-caspase 3 protein at each time point of brain death group were higher than those of the corresponding time point of sham group (P<0.05). Conclusions The relationship between brain death donor liver and cell apoptosis is closely related. Along with the extension of the brain death time in rabbits, the level of apoptosis of liver cells gradually increased, which affects the quality of liver donors after brain death.

Citation： ZHONGZi-biao, ZHANGMeng, XIONGYan, WANGYan-feng, YEQi-fa. The Quantitative Research in Hepatocyte Apoptosis Level from Rabbit Donors after Brain Death. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2016, 23(1): 22-27. doi: 10.7507/1007-9424.20160006 Copy

1.	Olson JC, Wendon JA, Kramer DJ, et al. Intensive care of the patient with cirrhosis. Hepatology, 2011, 54(5):1864-1872.
2.	郑树森, 徐骁. 积极推进中国肝移植的发展. 中华肝胆外科杂志, 2005, 11(7):437-439.
3.	Monbaliu D, Pirenne J, Talbot D. Liver transplantation using Dona-tion after Cardiac Death donors. J Hepatol, 2012, 56(2):474-485.
4.	Levesque E, Hoti E, Khalfallah M, et al. Impact of reversible cardiac arrest in the brain-dead organ donor on the outcome of adult liver transplantation. Liver Transpl, 2011, 17(10):1159-1166.
5.	郑树森, 吴健. 我国活体肝移植的现状与发展趋势. 外科理论与实践, 2008, 13(4):285-287.
6.	黄洁夫. 中国大陆肝移植的伦理与立法. 中华肝胆外科杂志, 2007, 45(5):292-296.
7.	Timek T, Vahl CF, Bonz A, et al. Triiodothyronine reverses depressed contractile performance after excessive catecholamine stimulation. Ann Thorae Surg, 1998, 66(5):1618-1625.
8.	Ranasinghe AM, Bonser RS. Endocrine changes in brain death and transplantation. Best Pract Res Clin Endocrinol Metab, 2011, 25(5):799-812.
9.	Li J, Yuan J. Caspases in apoptosis and beyond. Oncogene, 2008, 27(48):6194-6206.
10.	钟自彪, 叶啟发, 王彦峰, 等. 脑死亡状态下家兔肝脏质量评估新指标——线粒体醛脱氢酶. 中华肝胆外科杂志, 2013, 19(7):534-538.
11.	钟自彪, 王彦峰, 李玲, 等. 缓慢间断颅内加压法家兔脑死亡模型的建立及病理生理变化. 中华肝胆外科杂志, 2013, 19(8):534-538, 622-624.
12.	中华人民共和国国家科学技术委员会. 实验动物管理条例. 1988.
13.	Committee for the Update of the Guide for the Care and Use of Laboratory Animals; National Research Council. Guide for the Care and Use of Lab Animals. National Academies Press, 2010. http://www.nap.edu/catalog/12910.html.
14.	Quality Standards Subconmlittee of the Amecican Academy of Neurology. Practice parameters for determining brain death in adults (Summary statement). Neurology, 1995, 45(5):1012-1014.
15.	Kam PC, Ferch NI. Apoptosis:mechanisms and clinical implica-tions. Anaesthesia, 2000, 55(11):1081-1093.
16.	杨连君, 曹雪涛, 于益芝. Bcl-2,bax与肿瘤细胞凋亡. 中国肿瘤生物治疗杂志, 2003, 10(3):232-234.
17.	岳原亦, 张扬, 张一奇. Caspase家族与细胞凋亡. 中国医疗前沿, 2011, 6 (6):25-26.
18.	Li J, Yuan J. Caspases in apoptosis and beyond. Oncogene, 2008, 27(48):6194-6206.
19.	Salvesen GS, Ashkenazi A. Snapshot:caspases. Cell, 2011, 147(2):476-476.
20.	Ye H, Wang DP, Zhang CZ, et al. Pathological characteristics of liver allografts from donation after brain death followed by cardiac death in pigs. J Huazhong Univ Sci Technolog Med Sci, 2014, 34(5):687-691.
21.	Churchill EN, Disatnik MH, Mochy-Rosen D. Time-dependent and ethanol-induced cardiac protection from ischemia mediated by mitochondrial translocation of varepsilonPKC and activation of aldehyde dehydrogenase 2. J Mol Cell Cardiol, 2009, 46(2):278-284.
22.	Sun L, Ferreira JC, Mochly-Rosen D. ALDH2 activator inhibits increased myocardial infarction injury by nitroglycerin tolerance. Sci Transl Med, 2011, 3:(107):107-111.
23.	Li SY, Gomelsky M, Duan J, et al. Overexpression of aldehyde dehydrogenase-2 (ALDH2) transgene prevents acetaldehyde-induced cell injury in human umbilical vein endothelial cells:role of ERK and p38 mitogen-activated protein kinase. J Biol Chem, 2004, 279(12):11244-11252.
24.	Guo R, Zhong L, Ren J, et al. Overexpression of ALDH2 attenuates chronic alcohol exposure-induced apoptosis, change in Akt and Pim signaling in liver. Clin Exp Pharmacol Physiol, 2009, 36(5-6):463-468.
25.	Chen T, Cui J, Liang Y, et al. Identification of human liver mitochon-drial aldehyde dehydrogenase as a potential target for microcystin-LR. Toxicology, 2006, 220(1):71-80.
26.	Xu D, Guthrie JR, Mabry S, et al. Mitochondrial aldehyde dehydrogenase attenuates hyperoxia-induced cell death through activation of ERK/MAPK and PI3K-Akt pathways in lung epithelial cells. Am J Physiol Lung Cell Mol Physiol, 2006, 291(5):L966-L975.
27.	Lin MH, Yen JH, Wang CY, et al. Lipid peroxidation end product 4-hydroxy-trans-2-nonenal triggers unfolded protein response and heme oxygenase-1 expression in PC12 cells:Roles of ROS and MAPK pathways. Toxicology, 2014, 315:24-37.
28.	Lang XE, Wang X, Zhang KR, et al. Isoflurane preconditioning confers cardioprotection by activation of ALDH2. PLoS One, 2013, 8(2):e52469.

1. Olson JC, Wendon JA, Kramer DJ, et al. Intensive care of the patient with cirrhosis. Hepatology, 2011, 54(5):1864-1872.
2. 郑树森, 徐骁. 积极推进中国肝移植的发展. 中华肝胆外科杂志, 2005, 11(7):437-439.
3. Monbaliu D, Pirenne J, Talbot D. Liver transplantation using Dona-tion after Cardiac Death donors. J Hepatol, 2012, 56(2):474-485.
4. Levesque E, Hoti E, Khalfallah M, et al. Impact of reversible cardiac arrest in the brain-dead organ donor on the outcome of adult liver transplantation. Liver Transpl, 2011, 17(10):1159-1166.
5. 郑树森, 吴健. 我国活体肝移植的现状与发展趋势. 外科理论与实践, 2008, 13(4):285-287.
6. 黄洁夫. 中国大陆肝移植的伦理与立法. 中华肝胆外科杂志, 2007, 45(5):292-296.
7. Timek T, Vahl CF, Bonz A, et al. Triiodothyronine reverses depressed contractile performance after excessive catecholamine stimulation. Ann Thorae Surg, 1998, 66(5):1618-1625.
8. Ranasinghe AM, Bonser RS. Endocrine changes in brain death and transplantation. Best Pract Res Clin Endocrinol Metab, 2011, 25(5):799-812.
9. Li J, Yuan J. Caspases in apoptosis and beyond. Oncogene, 2008, 27(48):6194-6206.
10. 钟自彪, 叶啟发, 王彦峰, 等. 脑死亡状态下家兔肝脏质量评估新指标——线粒体醛脱氢酶. 中华肝胆外科杂志, 2013, 19(7):534-538.
11. 钟自彪, 王彦峰, 李玲, 等. 缓慢间断颅内加压法家兔脑死亡模型的建立及病理生理变化. 中华肝胆外科杂志, 2013, 19(8):534-538, 622-624.
12. 中华人民共和国国家科学技术委员会. 实验动物管理条例. 1988.
13. Committee for the Update of the Guide for the Care and Use of Laboratory Animals; National Research Council. Guide for the Care and Use of Lab Animals. National Academies Press, 2010. http://www.nap.edu/catalog/12910.html.
14. Quality Standards Subconmlittee of the Amecican Academy of Neurology. Practice parameters for determining brain death in adults (Summary statement). Neurology, 1995, 45(5):1012-1014.
15. Kam PC, Ferch NI. Apoptosis:mechanisms and clinical implica-tions. Anaesthesia, 2000, 55(11):1081-1093.
16. 杨连君, 曹雪涛, 于益芝. Bcl-2,bax与肿瘤细胞凋亡. 中国肿瘤生物治疗杂志, 2003, 10(3):232-234.
17. 岳原亦, 张扬, 张一奇. Caspase家族与细胞凋亡. 中国医疗前沿, 2011, 6 (6):25-26.
18. Li J, Yuan J. Caspases in apoptosis and beyond. Oncogene, 2008, 27(48):6194-6206.
19. Salvesen GS, Ashkenazi A. Snapshot:caspases. Cell, 2011, 147(2):476-476.
20. Ye H, Wang DP, Zhang CZ, et al. Pathological characteristics of liver allografts from donation after brain death followed by cardiac death in pigs. J Huazhong Univ Sci Technolog Med Sci, 2014, 34(5):687-691.
21. Churchill EN, Disatnik MH, Mochy-Rosen D. Time-dependent and ethanol-induced cardiac protection from ischemia mediated by mitochondrial translocation of varepsilonPKC and activation of aldehyde dehydrogenase 2. J Mol Cell Cardiol, 2009, 46(2):278-284.
22. Sun L, Ferreira JC, Mochly-Rosen D. ALDH2 activator inhibits increased myocardial infarction injury by nitroglycerin tolerance. Sci Transl Med, 2011, 3:(107):107-111.
23. Li SY, Gomelsky M, Duan J, et al. Overexpression of aldehyde dehydrogenase-2 (ALDH2) transgene prevents acetaldehyde-induced cell injury in human umbilical vein endothelial cells:role of ERK and p38 mitogen-activated protein kinase. J Biol Chem, 2004, 279(12):11244-11252.
24. Guo R, Zhong L, Ren J, et al. Overexpression of ALDH2 attenuates chronic alcohol exposure-induced apoptosis, change in Akt and Pim signaling in liver. Clin Exp Pharmacol Physiol, 2009, 36(5-6):463-468.
25. Chen T, Cui J, Liang Y, et al. Identification of human liver mitochon-drial aldehyde dehydrogenase as a potential target for microcystin-LR. Toxicology, 2006, 220(1):71-80.
26. Xu D, Guthrie JR, Mabry S, et al. Mitochondrial aldehyde dehydrogenase attenuates hyperoxia-induced cell death through activation of ERK/MAPK and PI3K-Akt pathways in lung epithelial cells. Am J Physiol Lung Cell Mol Physiol, 2006, 291(5):L966-L975.
27. Lin MH, Yen JH, Wang CY, et al. Lipid peroxidation end product 4-hydroxy-trans-2-nonenal triggers unfolded protein response and heme oxygenase-1 expression in PC12 cells:Roles of ROS and MAPK pathways. Toxicology, 2014, 315:24-37.
28. Lang XE, Wang X, Zhang KR, et al. Isoflurane preconditioning confers cardioprotection by activation of ALDH2. PLoS One, 2013, 8(2):e52469.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

The Quantitative Research in Hepatocyte Apoptosis Level from Rabbit Donors after Brain Death

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