血管紧张素转化酶2对肝细胞凋亡的影响

doi:10.3969/j.issn.1006-7795.2018.02.014

首都医科大学学报 ›› 2018, Vol. 39 ›› Issue (2): 223-229.doi: 10.3969/j.issn.1006-7795.2018.02.014

血管紧张素转化酶2对肝细胞凋亡的影响

宋丽妮^1,2, 曹曦^1,2, 刘薇¹, 杨金奎^1,2

1. 首都医科大学附属北京同仁医院内分泌科, 北京 100730;
2. 糖尿病防治研究北京市重点实验室, 北京 100730

收稿日期:2018-03-16 出版日期:2018-03-21 发布日期:2018-04-14
通讯作者: 杨金奎 E-mail:liliying@ccmu.edu.cn
基金资助:
国家自然科学基金（81670550），北京市属高等学校创新团队建设与教师职业发展计划项目（IDHT20150502）。

Effects of angiotensin converting enzyme 2 on apoptosis of hepatocytes

Song Lini^1,2, Cao Xi^1,2, Liu Wei¹, Yang Jinkui^1,2

1. Department of Endocrinology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;
2. Beijing Key Laboratory of Diabetes Research and Care, Beijing 100730, China

Received:2018-03-16 Online:2018-03-21 Published:2018-04-14
Supported by:
This study was supported by National Natural Science Foundation of China(81670550), Project of Construction of Innovative Teams and Teacher Career Development for Universities and Colleges Under Beijing Municipality (IDHT20150502).

摘要/Abstract

摘要： 目的探讨血管紧张素转化酶2（angiotensin converting enzyme 2，ACE 2）对肝细胞凋亡的影响及可能机制。方法 1利用ACE2过表达DNA载体转染人肝细胞株HepG2细胞；2MTT[3-（4，5-dimethyl-2-thiazolyl）-2，5-diphenyl-2H-tetrazoliumbromide]法检测过表达ACE2基因的HepG2细胞在棕榈酸（palmitate，PA）处理下的细胞活力；3原位DNA末端酶标记技术（terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay，TUNEL）检测同周龄雄性C57BL/6和ACE2基因敲除（ACE2^-/y）小鼠肝细胞凋亡情况；4实时荧光定量PCR（real-time PCR，RT-PCR）法检测凋亡相关基因的表达；5蛋白质印迹法（Western blot）检测B细胞淋巴瘤/白血病-2蛋白（anti-apoptotic protein B cell lymphoma-2，Bcl-2）、促凋亡基因（Bcl-2 associated X protein，Bax）、C/EBP同源蛋白（C/EBP homologous protein，CHOP）、C-Jun氨基末端激酶（C-Jun NH2-terminal kinase，JNK）和含半胱氨酸的天冬氨酸蛋白水解酶3（cysteinyl aspartate specific proteinase-3，caspase3）蛋白水平的表达情况。结果 1以不同浓度PA处理HepG2细胞24 h后表现为细胞活力的降低，而过表达ACE2基因组细胞活力明显高于绿色荧光蛋白（green fluorescent protein，GFP）组（PA浓度为0.4、0.8、1.0 mmol/L时，P<0.05）；2同周龄雄性ACE2^-/y小鼠肝脏细胞TUNEL阳性细胞明显多于对照组C57BL/6小鼠；3ACE2过表达显著降低了凋亡相关蛋白Bax，caspase-3，CHOP，磷酸化JNK的表达，而抗凋亡基因Bcl-2表达升高。与此同时，ACE2过表达也明显降低了内质网应激（endoplasmic reticulum stress，ERS）相关细胞凋亡通路基因的表达。结论 ACE2减少了肝细胞的凋亡，其机制可能与ACE2对肝脏内质网应激的保护作用有关。

关键词: 血管紧张素转换酶2, 内质网应激, 细胞凋亡

Abstract: Objective To identify the role of angiotensin-converting enzyme 2 (ACE2) in apoptosis of hepatic cells and to characterize the related mechanisms. Methods 1 HepG2 cells were transfected with ACE2 over expression plasmid DNA; 2MTT assay was used to measure HepG2 cells viability which over expressed ACE2 gene and then exposed to palmitic acid; 3 TUNEL assay was used to detect the apoptosis of hepatocytes in age-matched male C57BL/6 and ACE2^-/y mice; 4The expression of apoptosis related genes were detected by RT-PCR; 5Western blotting was used to study the expression of Bcl2, Bax, CHOP, phosphorylation-JNK and caspase3. Results 1We determined that exposure to different concentration of PA for 24 h caused a decrease in cell viability. However, the cell viability in ACE2-overexpressing HepG2 cells was significantly higher than GFP group (when the concentration of PA was 0.4, 0.8 or 1.0 mmol/L, P<0.05). 2The TUNEL positive hepatocytes in age-matched male ACE2^-/y mice group were more than those of C57BL-6 mice in the control group. 3In ACE2-overexpressing HepG2 cells, the relative protein expressions of Bax, caspase-3, CHOP and phosphorylation-JNK were decreased. While, the expression of Bcl2 was increased significantly. At this stage, over-expression of ACE2 also down-regulated the expression of ER-stress related apoptosis pathway genes. Conclusion ACE2 could be involved in anti-apoptotic via resistance to endoplasmic reticulum stress.

Key words: angiotensin-converting enzyme 2, endoplasmic reticulum stress, apoptosis

中图分类号:

R587.1

宋丽妮, 曹曦, 刘薇, 杨金奎. 血管紧张素转化酶2对肝细胞凋亡的影响[J]. 首都医科大学学报, 2018, 39(2): 223-229.

Song Lini, Cao Xi, Liu Wei, Yang Jinkui. Effects of angiotensin converting enzyme 2 on apoptosis of hepatocytes[J]. Journal of Capital Medical University, 2018, 39(2): 223-229.

参考文献

[1] Jia Y, Xi Z, Yu X, et al. Renin-angiotensin system activation accelerates atherosclerosis in experimental renal failure by promoting endoplasmic reticulum stress-related inflammation[J]. Int J Mol Med, 2017, 39(3):613.
[2] Kroy D C, Fabienne S, Pierluigi R, et al. Hepatocyte specific deletion of c-Met leads to the development of severe non-alcoholic-steatohepatitis in mice[J]. J Hepatol, 2014, 61(4):883-890.
[3] 史婷婷, 杨芳远, 曹曦,等. 血管紧张素转化酶2减少胰岛细胞的氧化应激及凋亡[J]. 首都医科大学学报, 2017,38(2):151-155.
[4] Ji Y, Gao F, Sun B, et al. Angiotensin-converting enzyme 2 inhibits apoptosis of pulmonary endothelial cells during acute lung injury through suppressing SMAD2 phosphorylation[J]. Cell Physiol Biochem, 2015, 37(2):759-767.
[5] 林振浩, 倪云杰, 侯良磊,等. 替米沙坦通过上调大鼠视网膜ACE2-Ang-(1-7)-Mas轴抑制细胞凋亡[J]. 中华心血管病杂志, 2015, 43(7):625-630.
[6] 晋学庆, 王前胜, 卢卓强,等. 血管紧张素转化酶2基因过表达改善人脐静脉内皮细胞生长[J]. 福建医科大学学报, 2012, 46(5):310-314.
[7] 陈旭. ACE2基因过表达对糖尿病心肌病内质网应激介导的心肌细胞凋亡的研究[D]. 济南:山东大学, 2014.
[8] Jung T W, Hwang H J, Hong H C, et al. Resolvin D1 reduces ER stress-induced apoptosis and triglyceride accumulation through JNK pathway in HepG2 cells[J]. Mol Cell Endocrinol, 2014, 391(1-2):30-40.
[9] Bechmann L P, Hannivoort R A, Gerken G, et al. The interaction of hepatic lipid and glucose metabolism in liver diseases[J]. J Hepatol, 2012, 56(4):952-964.
[10] Gentile C L, Frye M, Pagliassotti M J. Endoplasmic reticulum stress and the unfolded protein response in nonalcoholic fatty liver disease[J]. Antioxid Redox Signal, 2011, 15(2):505-521.
[11] Degracia D J, Kumar R, Owen C R, et al. Molecular pathways of protein synthesis inhibition during brain reperfusion|[colon]|Implications for Neuronal Survival or Death[J]. J Cerebral Blood Flow Metab, 2002, 22(2):127-141.
[12] Xu X, Liu T, Zhang A, et al. Reactive oxygen species-triggered trophoblast apoptosis is initiated by endoplasmic reticulum stress via activation ofcCaspase-12, CHOP, and the JNK pathway in toxoplasma gondii infection in mice[J]. Infect Immun, 2015, 83(4):1735.
[13] 吴涛, 季光, 郑培永,等. 内质网应激与肝细胞凋亡[J]. 世界华人消化杂志, 2007, 15(23):1569-1576.
[14] Zhu B, Zhang L, Zhang Y, et al. DNase I aggravates islet β-cell apoptosis in type 2 diabetes[J]. Mol Med Rep, 2016, 13(6):4577.
[15] Hetz C, Chevet E, Harding H P. Targeting the unfolded protein response in disease[J]. Nat Rev Drug Dis, 2013, 12(9):703-719.

血管紧张素转化酶2对肝细胞凋亡的影响

Effects of angiotensin converting enzyme 2 on apoptosis of hepatocytes

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	安松林, 马玲, 赵月, 李雁. 主要促进因子超家族成员MFSD2A对肝癌细胞增殖、凋亡和侵袭的影响[J]. 首都医科大学学报, 2021, 42(6): 1000-1006.
[2]	杨楠, 丁锚, 闫峰, 师文娟, 黄语悠, 赵咏梅. 远隔缺血预适应对脑缺血再灌注损伤大鼠缺血半暗带区PERK/p-eIF2α通路及自噬的影响[J]. 首都医科大学学报, 2021, 42(2): 225-231.
[3]	黄语悠, 房亚兰, 师文娟, 刘克建, 赵咏梅, 丁锚, 罗玉敏. 一氧化氮清除剂Carboxy-PTIO对短暂性局灶性脑缺血大鼠PERK/p-eIF2α通路的调节作用[J]. 首都医科大学学报, 2019, 40(5): 709-714.
[4]	张怡尘, 曹曦, 杨金奎. G蛋白偶联受体MAS对肝脏糖脂代谢的影响[J]. 首都医科大学学报, 2019, 40(3): 409-416.
[5]	卢晶, 沈涵, 程呈, 刘敬怡, 朱晓蓉, 谢荣荣, 袁明霞, 杨金奎. 人ether-α-go-go钾通道减少肝脏的内质网应激和凋亡[J]. 首都医科大学学报, 2019, 40(1): 45-52.
[6]	鹿星梦, 信中. Ang(1-7)通过调节内质网应激改善骨骼肌脂代谢及胰岛素抵抗[J]. 首都医科大学学报, 2018, 39(4): 532-536.
[7]	穆凌云, 李金霞, 张秋云, 陈煜, 高连印, 杜宇琼. 截断逆挽方对慢加急性肝衰竭模型大鼠血清TNF-α、肝组织p-JNK及c-Jun的影响[J]. 首都医科大学学报, 2017, 38(2): 282-288.
[8]	王梦然, 张健, 梁艳红, 陈艳蓉, 李敏, 肖瑶, 宿慧. 缺血后处理对老年大鼠心肌缺血再灌注损伤的保护作用[J]. 首都医科大学学报, 2015, 36(1): 132-136.
[9]	曹曦, 杨芳远, 史婷婷, 谢荣荣, 信中, 杨金奎. ACE2/Ang-(1-7)改善肝细胞糖代谢[J]. 首都医科大学学报, 2014, 35(6): 755-759.
[10]	叶娜, 董晓英, 李冬华. 卵巢早衰的颗粒细胞凋亡机制研究进展[J]. 首都医科大学学报, 2014, 35(3): 379-383.
[11]	王鹏雁;安建多;王池;王学从;武鑫瑞;王学江. 槲芪散及槲寄生提取物对肝癌细胞内游离钙离子浓度的影响[J]. 首都医科大学学报, 2012, 33(3): 291-296.
[12]	廖秋菊;王晶;秦俭. 胰岛素防治大鼠一氧化碳中毒迟发性脑病的实验研究[J]. 首都医科大学学报, 2012, 33(2): 233-237.
[13]	王燕;李萍;林燕;赵京霞;刘欣;何薇;梁代英;罗晓琴;王笑民. 鸡血藤黄酮类有效部位对人非小细胞肺癌A549细胞凋亡与线粒体膜电位的影响[J]. 首都医科大学学报, 2011, 32(6): 811-815.
[14]	丁月霞;;张丽;叶翠飞;王文;李林. 山茱萸环烯醚萜苷对穹隆海马伞切断大鼠海马区神经元存活和细胞凋亡调控因子的影响[J]. 首都医科大学学报, 2011, 32(1): 73-78.
[15]	闫颖;赵咏梅;赵志炜;王玉兰;李森. 内质网应激通路相关分子GRP78及CHOP在糖尿病脑病小鼠海马表达的变化[J]. 首都医科大学学报, 2011, 32(1): 90-94.