小鼠肝再生过程中MnSOD作用的初步研究

doi:10.3969/j.issn.1004-616x.2019.01.011

癌变·畸变·突变 ›› 2019, Vol. 31 ›› Issue (1): 58-63,78.doi: 10.3969/j.issn.1004-616x.2019.01.011

小鼠肝再生过程中MnSOD作用的初步研究

李鸽¹, 王耀斐¹, 干娜¹, 贺琪¹, 王冠¹, 韩占锋¹, 海春旭², 蔺兆星¹

1. 陕西省地方病防治研究所皮肤性病防治研究室, 陕西西安 710003;
2. 空军军医大学军事预防医学系毒理学教研室, 陕西省自由基生物学与医学重点实验室, 陕西西安 710032

收稿日期:2018-01-03 修回日期:2018-12-13 出版日期:2019-01-31 发布日期:2019-01-31
通讯作者: 蔺兆星,E-mail:1739423@qq.com E-mail:1739423@qq.com
作者简介:李鸽,E-mail:1016009751@qq.com。
基金资助:
国家自然科学基金项目（31670863，81270417，81573127）

Preliminary study on the role of MnSOD in liver regeneration

LI Ge¹, WANG Yaofei¹, GAN Na¹, HE Qi¹, WANG Guan¹, HAN Zhanfeng¹, HAI Chunxu², LIN Zhaoxing¹

1. Department of Dermatology and Sexually Transmitted Disease Prevention and Control, Shaanxi Provincial Institute for Endemic Disease Control, Xi'an 710003;
2. Department of Toxicology, School of Public Health, Medical University of the Air Force, Shaanxi Key Lab of Free Radical Biology and Medicine, Xi'an 710032, Shaanxi, China

Received:2018-01-03 Revised:2018-12-13 Online:2019-01-31 Published:2019-01-31

摘要/Abstract

摘要： 目的：研究肝切除术后小鼠肝再生过程中锰超氧化物歧化酶（MnSOD）的表达及其活性的变化，探讨MnSOD在肝再生中的作用。方法：采用经典小鼠肝切除模型，将38只雄性BALB/c小鼠，随机分为30%肝切除组（30% PH组）18只，70%肝切除组（70% PH组）18只，以及对照组2只。2个肝切除组分别于术后6 h和1、2、3、5、7 d这6个时间点随机各抽取3只小鼠处死，对照组小鼠在行假手术后即处死。取肝组织，制备冰冻切片使用DHE染色法在激光共聚焦显微镜下检测活性氧（ROS）水平，实时荧光定量PCR检测小鼠肝组织中MnSOD mRNA表达水平，Western blot检测小鼠肝组织中MnSOD蛋白的表达水平，采用MnSOD试剂盒检测小鼠肝组织中的MnSOD活性。结果：肝切除术后，与对照组相比，30% PH组小鼠肝组织ROS水平在术后6 h和1 d增加，MnSOD mRNA表达水平增加（P < 0.05），MnSOD蛋白含量无显著变化（P > 0.05）；MnSOD的活性在术后第1和2天较高，第3和5天较低，第7天恢复。70% PH组小鼠肝组织ROS水平在术后第1~5天均升高，MnSOD mRNA的水平先下降后逐渐恢复，MnSOD的蛋白含量降低，MnSOD的活性在术后第6小时和1天增加，第2~7天均处于降低状态（P均 < 0.05）。结论：在肝切除术后小鼠的肝再生过程迅速启动，尤其是在70% PH后肝细胞迅速增殖，并在术后一段时间逐渐恢复到静息状态，其机制可能与MnSOD含量和活性的下调从而导致ROS升高有关。

关键词: 锰超氧化物歧化酶, 肝切除, 肝再生, 活性氧, 细胞增殖

Abstract: OBJECTIVE:The aim of this study was to investigate changes of MnSOD expression and its activity in liver regeneration after partial hepatectomy. METHODS:Using the classic mouse partial hepatectomy model,38 male BALB/c mice were randomly divided into 30% partial hepatectomy group (30%PH) and 70% partial hepatectomy group (70%PH). There were 18 mice in each experimental group,and 2 as sham operation control group. 3 mice was randomly selected from the two hepatectomy groups at 6 h,1 d,2 d,3 d,5 d and 7 d after the operation,and the control group was executed immediately after the sham operation. Liver tissues in the PH groups were harvested at 6 h,1 d,2 d,3 d,5 d and 7 d respectively. Frozen sections from collected tissues were used to detect the level of reactive oxygen species by the DHE dyeing method and using laser confocal microscopy. MnSOD mRNA was measured by Real Time Q-PCR. Expression of MnSOD protein was determined by Western blot,and activity of MnSOD was assessed using the MnSOD kit. RESULTS:Compared with the control group,the ROS levels in liver tissues from PH mice increased at 6 h and 1 d after operation,expression level of MnSOD mRNA increased (P < 0.05),but protein content of MnSOD did not change significantly in the 30%PH group (P > 0.05). Activities of MnSOD was higher on 1 d and 2 d,lower on 3 d and 5 d,and then recovered on 7 d after 30%PH. In the 70%PH group,ROS level in liver tissues increased from 1 d to 5 d after operation; MnSOD mRNA levels decreased first and then gradually recovered; protein content of MnSOD decreased; and activities of MnSOD increased at 6 h and 1 d,and then decreased during 2 d to 7 d (all P < 0.05). CONCLUSION:Liver regeneration started rapidly after hepatectomy in mice,especially after 70%PH. The hepatocytes rapidly proliferated,and then gradually returned to a resting state after a period of time. The observed changes might be related to the down-regulation of MnSOD content and activity,which led to the increase of ROS.

Key words: MnSOD, partial hepatectomy, liver regeneration, reactive oxygen species, cell proliferation

中图分类号:

R364.3

李鸽, 王耀斐, 干娜, 贺琪, 王冠, 韩占锋, 海春旭, 蔺兆星. 小鼠肝再生过程中MnSOD作用的初步研究[J]. 癌变·畸变·突变, 2019, 31(1): 58-63,78.

LI Ge, WANG Yaofei, GAN Na, HE Qi, WANG Guan, HAN Zhanfeng, HAI Chunxu, LIN Zhaoxing. Preliminary study on the role of MnSOD in liver regeneration[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 58-63,78.

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小鼠肝再生过程中MnSOD作用的初步研究

Preliminary study on the role of MnSOD in liver regeneration

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