神经生长因子对H9c2心肌细胞缺氧/复氧后凋亡的保护作用及其机制

被引:15
作者
谢飞
魏珂
闵苏
郝学超
朱贤林
机构
[1] 重庆医科大学附属第一医院麻醉科
关键词
H9c2心肌细胞; 缺氧/复氧损伤; 神经生长因子; PI3k-Akt通路; 心肌细胞存活率; 心肌细胞凋亡率; 内质网应激蛋白;
D O I
暂无
中图分类号
R541 [心脏疾病];
学科分类号
1002 ; 100201 ;
摘要
目的探讨神经生长因子(NGF)对H9c2心肌细胞缺氧/复氧(H/R)后凋亡的保护作用及其机制。方法将培养的H9c2心肌细胞随机分为5组:正常对照组(C组)、缺氧/复氧组(H/R组)、NGF组(N组)、NGF+PI3k受体特异性阻断剂LY294002组(N+L组)和LY294002组(L组)。C组用正常培养液在通有95%空气+5%CO2的37℃培养箱内正常培养8 h;H/R组用预先经95%N2+5%CO2的混合气饱和1 h的模拟缺氧液置换正常培养液后,放入通有95%N2+5%CO2的37℃培养箱缺氧培养4 h,然后将模拟缺氧液换用预先经95%空气+5%CO2饱和的正常培养液,在正常条件下复氧4 h;N组、N+L组和L组先缺氧培养4 h(缺氧条件同H/R组),再分别用预先经95%空气+5%CO2饱和的含NGF、NGF+LY294002和LY294002的正常培养液置换模拟缺氧液,在正常条件下复氧4 h。NGF和LY294002的终浓度分别为0.05 mg·L-1和3 mg·L-1。CCK-8比色法测定各组细胞的存活率;应用碘化丙啶(PI)染色流式细胞术检测细胞凋亡率;Western blot法检测胞内Akt磷酸化水平及内质网应激蛋白Caspase-12蛋白的表达。结果 N组与N+L组或H/R组或L组相比能明显提高H9c2心肌细胞经H/R损伤后的存活率,减少细胞凋亡率,上调Akt磷酸化水平,降低内质网应激蛋白Caspase-12表达;LY294002明显抑制了NGF对Akt磷酸化水平的上调作用。结论 NGF对经H/R损伤的H9c2心肌细胞有抗凋亡作用,其机制可能与PI3k-Akt通路有关。
引用
收藏
页码:506 / 510
页数:5
相关论文
共 10 条
[1]   PI3K/Akt信号通路在硫化氢保护PC12细胞对抗化学性缺氧损伤的作用 [J].
孟金兰 ;
陈雅嘉 ;
陈红 ;
董艳芬 ;
邢德刚 ;
梁燕玲 ;
兰爱平 ;
冯鉴强 .
中国药理学通报, 2013, 29 (02) :257-261
[2]   缺氧复氧诱导大鼠心肌细胞内质网应激反应 [J].
马青变 ;
高炜 ;
郭艳红 ;
赵春玉 ;
薛林 ;
唐朝枢 .
北京大学学报(医学版), 2005, (04) :386-388
[3]   Nerve Growth Factor Promotes Cardiac Repair following Myocardial Infarction [J].
Meloni, Marco ;
Caporali, Andrea ;
Graiani, Gallia ;
Lagrasta, Costanza ;
Katare, Rajesh ;
Van Linthout, Sophie ;
Spillmann, Frank ;
Campesi, Ilaria ;
Madeddu, Paolo ;
Quaini, Federico ;
Emanueli, Costanza .
CIRCULATION RESEARCH, 2010, 106 (07) :1275-U230
[4]  
Nerve growth factor attenuates 2-deoxy- d -glucose-triggered endoplasmic reticulum stress-mediated apoptosis via enhanced expression of GRP78[J] . Soichiro Kishi,Koji Shimoke,Yosuke Nakatani,Takafumi Shimada,Nobuaki Okumura,Katsuya Nagai,Kazuo Shin-ya,Toshihiko Ikeuchi.Neuroscience Research . 2009 (1)
[5]   Nerve growth factor and brain-derived neurotrophic factor attenuate angiotensin-II-induced facilitation of calcium channels in acutely dissociated nucleus tractus solitarii neurons of the rat [J].
Endoh, Takayuki ;
Sato, Daisuke ;
Wada, Yoshiyuki ;
Ishihara, Kazuyuki ;
Hashimoto, Sadamitsu ;
Yoshinari, Masao ;
Matsuzaka, Kenichi ;
Tazaki, Masakazu ;
Inoue, Takashi .
ARCHIVES OF ORAL BIOLOGY, 2008, 53 (12) :1192-1201
[6]   δPKC participates in the endoplasmic reticulum stress-induced response in cultured cardiac myocytes and ischemic heart [J].
Qi, Xin ;
Vallentin, Alice ;
Churchill, Eric ;
Mochly-Rosen, Daria .
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY, 2007, 43 (04) :420-428
[7]   Endoplasmic reticulum stress gene induction and protection from ischemia/reperfusion injury in the hearts of transgenic mice with a tamoxifen-regulated form of ATF6 [J].
Martindale, Joshua J. ;
Fernandez, Rayne ;
Thuerauf, Donna ;
Whittaker, Ross ;
Gude, Natalie ;
Sussman, Mark A. ;
Glembotski, Christopher C. .
CIRCULATION RESEARCH, 2006, 98 (09) :1186-1193
[8]   NGF-induced phosphatidylinositol 3-kinase signaling pathway prevents thapsigargin-triggered ER stress-mediated apoptosis in PC12 cells [J].
Shimoke, K ;
Kishi, S ;
Utsumi, T ;
Shimamura, Y ;
Sasaya, H ;
Oikawa, T ;
Uesato, S ;
Ikeuchi, T .
NEUROSCIENCE LETTERS, 2005, 389 (03) :124-128
[9]   Aktion in the nucleus [J].
Webster, KA .
CIRCULATION RESEARCH, 2004, 94 (07) :856-859
[10]   Gene expression of neurotrophins and their receptors in cultured rat vascular smooth muscle cells [J].
Nemoto, E ;
Fukamachi, E ;
Nemoto, F ;
Miyata, S ;
Hamada, M ;
Nakamura, Y ;
Senba, E ;
Ueyama, T .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 245 (01) :284-288