Hepatocyte growth factor attenuates ischemia/reperfusion induced cardiomyocyte apoptosis via downregulating calcium sensing receptor expression
10.3760/cma.j.issn.0253-3758.2010.11.012
- VernacularTitle:肝细胞生长因子的抗鼠心肌细胞凋亡作用与抑制钙敏感受体有关
- Author:
Ling YAN
1
;
Tie-Bing ZHU
;
Lian-Sheng WANG
;
Zheng-Xian TAO
;
Shi-Yang PAN
;
Zhi-Jian YANG
;
Ke-Jiang CAO
Author Information
1. 南京医科大学附属第一医院
- Keywords:
Myocardial reperfusion injury;
Hepatocyte growth factor;
Receptors,calciumsensing;
Apoptosis
- From:
Chinese Journal of Cardiology
2010;38(11):1019-1024
- CountryChina
- Language:Chinese
-
Abstract:
Objective To examine whether the anti-apoptotic effect of hepatocyte growth factor (HGF) in cardiomyocytes underwent ischemia/reperfusion (I/R) is associated with downregulation of calcium sensing receptor (CaSR) mRNA expression. Methods Neonatal rat cardiomyocytes were isolated and randomly divided into 7 groups: control, I/R, GdCl3, GdCl3 + NiCl2 + CdCl2, GdCl3 + LY294002,GdCl3 + HGF, GdCl3 + HGF + LY294002. L/R was established by incubating primary neonatal rat ventricular cardiomyocytes in ischemia-mimetic solution for 2 h, then reincubated in normal culture medium for 24 h.Cardiomyocyte apoptosis was detected by TUNEL. The expression of CaSR mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). The expression of Caspase-3, Bcl-2 and Phosphoinositide-3 Kinase (PI3K) was analyzed by Western blot. Results I/R enhanced the expression of CaSR mRNA ( I/R: 2. 62 ± 0. 41, control: 1.00 ± 0. 31, P ＜ 0. 01 ) and cardiomyocyte apoptosis [ I/R:( 15.32 ± 2. 54) %, control: (2. 90 ± 1.45 ) %, P ＜ 0. 01 ]. GdCl3 further increased the expression of CaSR mRNA (GdCl3:4.46 ±0.62, I/R:2.62±0.41, P＜0.01) and cardiomyocyte apoptosis [GdCl3:(25.36 ±2. 60) %, I/R: ( 15.32 ± 2. 54) %, P ＜ 0. 01 ], along with upregulation of Caspase-3 ( GdCl3: 1.93 ± 0. 28,I/R:1. 50 ± 0. 21, P ＜ 0. 01 ), downregulation of Bcl-2 (GdCl3:0. 82 ± 0. 18, I/R: 1.71 ± 0. 30, P ＜ 0. 01 )and PI3 K phosphorylation inhibition ( I/R:0. 87 ± 0. 08, GdCl3:0. 61 ± 0. 07, P ＜ 0. 01 ). Combination of GdCl3 with LY294002 furthex enhanced cardiomyocytes apoptosis [ GdCl3 + LY294002: ( 32. 6 ± 3.42 ) %,GdCl3: (25.36 ± 2. 60) %, P ＜ 0. 01 ] but did not affect CaSR mRNA expression ( GdCl3 + LY294002:4. 27 ± 0. 56, GdCl3:4. 46 ± 0. 62, P ＞ 0. 05). HGF decreased I/R- and GdCl3-induced apoptosis [ GdCl3 +HGF: ( 11.8 ± 1.89 ) %, GdCl3: ( 25.36 ± 2. 60 ) %, P ＜ 0. 05 ] by suppressing Caspase-3 ( GdCl3 +HGF:1.12±0.23, (GdCl3:1.93 ±0.28, P＜0. 05;GdCl3 + HGF+LY294002:1.87±0.31,GdCl3 +LY294002:3.86 ± 0. 47, P ＜ 0. 05 ) and promoting Bcl-2 ( GdCl3 + HGF: 2. 56 ± 0. 54, GdCl3: 0. 82 ±0. 18, P＜0.05;GdCl3 + HGF + LY294002:1.68 ±0.28,GdCl3 + LY294002:0.68 ±0. 13, P＜0.05)and PI3K phosphoration expression (GdCl3 + HGF:2. 87 ±0. 21 ,GdCl3 :0. 61 ±0. 07, P ＜0. 05;GdCl3 +HGF+ LY294002:2.01 ± 0. 14, GdCl3 + LY294002:0.44 ± 0. 10, P ＜ 0.05) in accordance with downregulation of CaSR mRNA expression ( GdCl3 + HGF: 1.46 ± 0. 37, GdCl3:4. 46 ± 0. 62, P ＜ 0. 01 ).Conclusion HGF exerts protective role in I/R-induced apoptosis at least in part by inhibiting CaSR mRNA expression along with promoting Bcl-2, suppressing Caspase-3 expression and stimulating PI3K phosphorylation signaling pathway.
