Korean Circ J.  2017 Nov;47(6):949-959. 10.4070/kcj.2016.0353.

Cardioprotective Effect of the SDF-1α/CXCR4 Axis in Ischemic Postconditioning in Isolated Rat Hearts

• Kim JS
• Jang Y
• Kim JH
• Park YH
• Hwang SA
• Kim J
• Lee SR
• Xu Z
• Ban C
• Ahn K
• Chun KJ

• KMID: 2396488
• DOI: http://doi.org/10.4070/kcj.2016.0353

Abstract

BACKGROUND AND OBJECTIVES
Information about the role of the stromal cell-derived factor-1α (SDF-1α)/chemokine receptor type 4 (CXCR4) axis in ischemic postconditioning (IPOC) is currently limited. We hypothesized that the SDF-1α/CXCR4 signaling pathway is directly involved in the cardioprotective effect of IPOC.
METHODS
Isolated rat hearts were divided into four groups. The control group was subjected to 30-min of regional ischemia and 2-hour of reperfusion (n=12). The IPOC group was induced with 6 cycles of 10-second reperfusion and 10-second global ischemia (n=8) in each cycle. The CXCR4 antagonist, AMD3100, was applied before reperfusion in the IPOC group (AMD+IPOC group, n=11) and control group (AMD group, n=9). Hemodynamic changes with electrocardiography were monitored and infarct size was measured. The SDF-1α, lactate dehydrogenase (LDH) and creatine kinase (CK) concentrations in perfusate were measured. We also analyzed extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt phosphorylation state expression.
RESULTS
IPOC significantly reduced infarct size, but AMD3100 attenuated the infarct reducing effect of IPOC. IPOC significantly decreased LDH and CK, but these effects were reversed by AMD3100. ERK1/2 and Akt phosphorylation increased with IPOC and these effects were blocked by AMD3100.
CONCLUSION
Based on the results of this study, SDF-1α/CXCR4 signaling may be involved in IPOC cardioprotection and this signaling pathway couples to the ERK1/2 and Akt pathways.

Keyword

Reperfusion injury; Ischemic postconditioning; Stromal cell-derived factor-1α; CXCR4 receptors

MeSH Terms

Animals
Creatine Kinase
Electrocardiography
Family Characteristics
Heart*
Hemodynamics
Ischemia
Ischemic Postconditioning*
L-Lactate Dehydrogenase
Phosphorylation
Phosphotransferases
Rats*
Receptors, CXCR4
Reperfusion
Reperfusion Injury
Creatine Kinase
L-Lactate Dehydrogenase
Phosphotransferases
Receptors, CXCR4

Figure

• 

  Figure 1 Experimental protocol. CON group was subjected to 30 minutes of regional ischemia followed by 120 minutes of reperfusion. IPOC group was induced by 6 cycles of 10 seconds reperfusion and 10 seconds global ischemia immediately after reperfusion. CXCR4 antagonist AMD3100 was administered from 20 minutes before reperfusion to 30 minutes after reperfusion. CON = untreated control hearts (n=12); CXCR4 = chemokine receptor type 4; IPOC = ischemic postconditioning (n=8).

• 

  Figure 2 (A) Representative sequential LV slices from each group showing the AN (pale area) with TTC staining. (B) Percent of infarct area over AR. All data are expressed as mean±standard deviation. There were significant differences between p=0.021 vs. CON group and p=0.045 vs. AMD+IPOC group. AMD = AMD3100 treatment in CON (n=9); AMD+IPOC = CXCR4 antagonist AMD3100 treatment in IPOC (n=11); AN = area of necrosis; AR = area at risk; CON = untreated control hearts (n=12); IPOC = ischemic postconditioning (n=8); TTC = 2,3,5-triphenyltetrazolium chloride.

• 

  Figure 3 Coronary effluent LDH and CK measured at 60 minutes after reperfusion. Data are expressed as mean±standard deviation. p<0.050 vs. CON group. AMD = AMD3100 treatment in CON (n=9); AMD+IPOC = CXCR4 antagonist AMD3100 treatment in IPOC (n=11); CK = creatine kinase; CON = untreated control hearts (n=12); IPOC = ischemic postconditioning (n=8); LDH = lactate dehydrogenase.

• 

  Figure 4 Confocal image of representative immunofluorescent staining for SDF-1 (green), DAPI (blue, nuclear DNA), and merged image from isolated perfused rat hearts. AMD = AMD3100 treatment in CON (n=9); AMD+IPOC = CXCR4 antagonist AMD3100 treatment in IPOC (n=11); CON = untreated control hearts (n=12); DAPI = 4',6-diamidino-2-phenylindole; IPOC = ischemic postconditioning (n=8); SDF-1 = stromal cell-derived factor-1.

• 

  Figure 5 (A) Representative western blotting for p-Akt and t-Akt intensity. (B) Representative western blotting for p-ERK1/2 and t-ERK1/2 intensity. Bar graph shows percentage change in ERK1/2 and Akt phosphorylation relative to CON group. Increased phosphorylation of Akt and ERK1/2 in IPOC-induced hearts was totally blocked by AMD3100. Data are expressed as mean±standard deviation. p<0.050 vs. CON group. AMD = AMD3100 treatment in CON (n=9); AMD+IPOC = CXCR4 antagonist AMD3100 treatment in IPOC (n=11); CON = untreated control hearts (n=12); IPOC = ischemic postconditioning (n=8); p-Akt = phospho-Akt; p-ERK1/2 = phospho-extracellular signal-regulated kinase 1/2; t-Akt = total-Akt; t-ERK1/2 = total-extracellular signal-regulated kinase 1/2.

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