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1、 AMPK inhibition of HO-1 in cardiac hypertrophy Of: Chenguang Qin, Liu Chen and Zhang Chengxi, Meng Rong Sen, Chen Baolin, Xiongzhao Jun, Dong Calls Abstract Objective AMP-activated protein kinase (AMPK) and heme oxygenase HO-1 activation on myocardial hypertrophy, and HO-1 inhibition of cardiac hyp
2、ertrophy in AMPKs role. Method in cultured neonatal rat cardiac cells were studied by Western blot hybridization detection of 5 - amino -4 imidazole carboxamide ribonucleotide (AICAR) on AMPK phosphorylation and the extent of the impact of HO-1, and of myocardial cells 3H - leucine incorporation and
3、 cell surface area was observed HO-1 AMPK and myocardial hypertrophy. results AICAR on AMPK and the role of HO-1 are activated, AICAR blocked the cardiac hypertrophy induced by Ang effect; blocking the activation of HO-1 is able to decrease the inhibitory effect of AICAR. Conclusion AMPK can inhibit
4、 cardiac hypertrophy, AMPK can activate HO-1, HO-1 in myocardial hypertrophy of the mechanism of AMPK plays an important role. Keywords: AMPK; HO-1; cardiac hypertrophy Abstract: Objective To investigate the effect of AMPK and HO-1 activation on hypertrophic cardiac myocyte, and the effect of HO-1 i
5、n the mechanism of AMPK inhibition of cardiomyocyte hypertrophy. Methods Cultured rat neonatal cardiacmyocytes were incubated with Ang to induce cardiac hypertrophy. Cardiac myocyte surface area and 3H leucine incorporation was measured to determine the effect of AMPK and HO-1 on cardiac hypertrophy
6、. The effect of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) on adenosine monophosphate activated protein kinase (AMPK) phosphorylation and HO-1 activaton were detected by Western blot. Results Compared with control group, activated AMPK significantly enhanced the transcriptive activity of
7、AMPK, and increased HO-1 protein expression in cardiac myocyte. The increasement of cell surface area by Ang was significantly inhibited after treatment of AICAR. However, this inhibitory effect was attenuated by HO-1 inhibitor. Conclusion AMPK can attenuate the cardiac hypertrophy induced by Ang ,
8、AMPK can upregulate the HO-1 in cardiac hypertrophy induced by AngII. HO-1 play a significant role in the effect of the AMPK on inhibiting cardiac hypertrophy. Adenosine monophosphate-activated protein kinase (AMP-activated protein kinase, AMPK) is a eukaryotic cell biology in the widespread presenc
9、e of serine / threonine protein kinase, a major regulator of metabolism and energy. AMPK by the , and subunits, which the 1 and 2 subtypes are mainly distributed in the myocardium 1-3, myocardial cells AMPK activation and myocardial ischemia, oxidative stress and some stimuli such as catecholamines
10、and angiotensin on 4-6, recent studies have shown that, AMPK could inhibit protein synthesis in cardiac myocytes reduces myocardial hypertrophy 7. HO-1 is the heme oxygenase (HO) isozymes, one can be more kinds of stimuli, such as: ischemia reperfusion, heat shock, such as heavy metals and hypoxia i
11、nduced increase. HO-1 as a novel myocardial protective factor, the product through its anti-inflammatory, antioxidant, anti-apoptotic and anti- role in cardiovascular disease, arrhythmia, etc. play an important protective role 8-11, some research results in recent years, HO-1 can inhibit the role of
12、 cardiac hypertrophy 12-13. AMPK and HO-1 based on inhibit cardiac hypertrophy in both play an important role, both of which can be ischemia, such as angiotensin-induced oxidative stress and increased expression of AMPK and a number of downstream factors such as p38MAPK, NO, etc. involved in the act
13、ivation of HO-1 14-16. This suggests that the two may exist a relationship between. To test this hypothesis, this study by observing the AMPK agonist AICAR on AMPK activation and the role of HO-1, and AMPK, HO-1 and HO- 1 blocker zinc protoporphyrin (ZnPP ) on myocardial hypertrophy, myocardial cell
14、s to confirm that HO-1 in myocardial hypertrophy in AMPK plays an important role in the mechanism, trying from a new perspective of AMPK on cardiac hypertrophy role. 1 Materials and methods 1.1 Materials and reagents Ang (Anaspec); AICAR (Cell sinaling); Compound C (Merk), 3H-Leucine (Beijing Atomic
15、 Energy Research Institute, radioactive concentration of 3.7 107 Bq / mL); ready-to-immunohistochemistry kit (new step company), ZnPP (Ssigma); DMEM F12 medium and newborn calf serum (Hyclone); HO-1 antibody (Abcam); Phospho-AMPK-alpha (Thr172) antibody, AMPK-alpha antibody (CST), GADPH (Shanghai Ka
16、ng Cheng ), sheep anti-mouse and goat anti-rabbit IgG-HRP (Wuhan Boster), BCA-200 protein quantification kit (Pierce); PVDF membrane (Millipower); ECL (Pik days). 1.2 neonatal rat cardiomyocytes in primary culture 1 2 d SD newborn rat ventricular tissue cut into pieces (1 mm3 size). To 0.5 g / L collagenase digestion. Thermostatic bath at 37 gently oscillating 2 h, then abandoned to the suspensi
