Danqi Pill Protects Against Heart Failure Post-Acute Myocardial Infarction via HIF-1α/PGC-1α Mediated Glucose Metabolism Pathway

Abstract

Aim: Heart failure (HF) post-acute myocardial infarction (AMI) leads to a large number of hospitalizations and deaths worldwide. Danqi pill (DQP) is included in the 2015 national pharmacopoeia and widely applied in the treatment of HF in clinics in China. We examined whether DQP acted on glucose metabolism to protect against HF post-AMI via hypoxia inducible factor-1 alpha (HIF-1α)/peroxisome proliferator-activated receptor α co-activator (PGC-1α) pathway.

Methods and results: In this study, left anterior descending (LAD) artery ligation induced HF post-AMI rats and oxygen-glucose deprivation-reperfusion (OGD/R)-induced H9C2 cell model were structured to explore the efficacy and mechanism of DQP. Here we showed that DQP protected the heart against ischemic damage as evidenced by improved cardiac functions and attenuated inflammatory infiltration. The expressions of critical proteins involved in glucose intake and transportation such as GLUT4 and PKM2 were up-regulated, while negative regulatory proteins involved in oxidative phosphorylation were attenuated in the treatment of DQP. Moreover, DQP up-regulated NRF1 and TFAM, promoted mitochondrial biogenesis and increased myocardial adenosine triphosphate (ATP) level. The protection effects of DQP were significantly compromised by HIF-1α siRNA, suggesting that HIF-1α signaling pathway was the potential target of DQP on HF post-AMI.

Conclusions: DQP exhibits the efficacy to improve myocardial glucose metabolism, mitochondrial oxidative phosphorylation and biogenesis by regulating HIF-1α/PGC-1α signaling pathway in HF post-AMI rats.

Keywords: Danqi Pill (DQP); HIF-1α/PGC-1α pathway; acute myocardial infarction (AMI); glucose metabolism; heart failure (HF).

Figure 1

Danqi Pill (DQP) in different doses reduced cardiac dysfunctions, inhibited inflammatory cell infiltration and increased adenosine triphosphate (ATP) production. (A) Representative echocardiograms in heart sections. (B) Echocardiography analyses of ejection fraction (EF), fractional shortening (FS), left ventricular internal dimension-diastole (LVID;d), and left ventricular internal dimension-systole (LVID;s). (C) Histological analyses of H&E staining. The scale bar is 50 μm. (D) Myocardial ATP levels in different groups. ^# P < 0.05, ^## P < 0.01, ^###P<0.001 compared with sham; ^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 compared with model. N = 6 per group.

Figure 2

Danqi Pill (DQP) promoted cardiac glucose metabolism in heart failure (HF) post-acute myocardial infarction (AMI) rats. (A) ¹⁸F-FDG uptake detected by PET/CT in four groups. Quantitative analyses of mean, max, and min standardized uptake value (SUV). (B) Cardiac protein expressions of GLUT4 and PKM2 and densitometric analyses. (C) Immunostaining of GLUT4 in cardiac sections and quantification of GLUT4. Region of interest (ROI) = 12 per rat. ^#P < 0.05, ^###P < 0.001 compared with sham; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 compared with model. N = 3 per group for western blotting. N = 6 per group for IHC.

Figure 3

Danqi Pill (DQP) promoted glucose metabolism by activating myocardial mitochondrial oxidation pathway in heart failure (HF) post-acute myocardial infarction (AMI) rats. (A) Western blot analysis the expressions of PDK4, PDK1, and UCP2 in all groups. (B) Western blot analysis the expressions of NRF1 and mitochondrial transcription factor A (TFAM) in all groups. Densitometric analysis was shown in the graph. GADPH was used as internal reference. ^##P < 0.01, ^###P < 0.001 compared with sham; ^*P < 0.05, ^**P < 0.01,^***P < 0.001 compared with model. N = 3 per group.

Figure 4

Danqi Pill (DQP) activated HIF1α/PGC1α signaling pathway in heart failure (HF) post-acute myocardial infarction (AMI) rats. (A) Western blot analysis the expressions of PPARα, RXRα, and PGC-1α in the four groups. Densitometric analysis was showed. (B) Effects of DQP on expressions of HIF-1α and PHD2 in HF post-AMI Rats; WB bands and protein quantitative results of HIF-1α and PHD2 in heart tissues of rats. ^# p < 0.05, ^## p < 0.01, ^###P < 0.001 compared with sham; ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 compared with model. N =3 per group.

Figure 5

Danqi Pill (DQP) protected against oxygen-glucose deprivation-reperfusion (OGD/R)-induced H9C2 cells injury via up-regulating HIF-1α. (A) Immunoblot of HIF-1α in H9C2 Cells. (B) HIF-1α small interfering RNA (siRNA) suppressed the facilitation of DQP in the proliferation of OGD/R H9C2 cells. (C) HIF-1α siRNA decreased mitochondrial membrane potential (△Ψm) with/without DQP treatment in OGD/R-induced H9C2 cells. △Ψm was measured by JC-1 probe (400 ×, Scale bar 100 μm). (D) Intracellular adenosine triphosphate (ATP) levels in H9C2 Cells. ^* P < 0.05, ^** P < 0.01, ^*** P < 0.01 compared with model; ^### P < 0.001 compared with DQP. N = 3 per group for WB and N=6 for the other experiments.

Figure 6

Danqi Pill (DQP) targeted on HIF-1α/PGC-1α signaling pathway and finally elevated cardiac energy metabolism in the failing heart.