Korean Circ J.  2008 Jul;38(7):360-365. 10.4070/kcj.2008.38.7.360.

Detection of Transferrin Receptor Over-Expression in a Rodent Model of Myocardial Ischemia/Reperfusion Injury Using Novel 99mTc Transferrin Conjugates: A Pilot Study

• Jeong HJ
• Kim EM
• Kim NH
• Lee CM
• Cheong SJ
• Kim DW
• Lim ST
• Yoon KJ
• Nah JW
• Sohn MH

• KMID: 2225769
• DOI: http://doi.org/10.4070/kcj.2008.38.7.360

Abstract

BACKGROUND AND OBJECTIVES: Reperfusion of ischemic myocardium is necessary to salvage tissue from eventual death. However, new pathophysiological changes are initiated after reperfusion. The aim of this study was to investigate one of the mechanisms of ischemia/reperfusion (I/R) injury, and we focused on transferrin.
MATERIALS AND METHODS
Male Spragre-Dawley (SD) rats were used for the I/R model. Myocardial ischemia was produced by occlusion of the left anterior descending coronary artery for 30 minutes. 99mTc Transferrin-Chitosan-hydrazino nicotinate hydrochloride (HYNIC) (Tfc) (/37 MBq/mL) was injected once after the reperfusion was finished. Autoradiography, hematoxylin and eosin (H & E) staining and determination of the tissue myeloperoxidase (MPO) activity were performed.
RESULTS
Autoradiography showed remarkable 99mTc-Tfc uptake in the left ventricular myocardium at the reperfusion period from 0 to 1.5 hours, whereas no uptake was demonstrated at 3 hours. The uptake was increased again at 6 and 24 hours. Western blotting showed that the transferrin receptor (TfR) proteins were increased at 0 to 1.5 hours compared with that of the control; this expression of TfR disappeared at 3 hours, and it showed up for the second time at 6 and 24 hours. The MPO activity only at 24 hours was significantly higher than that of the control and those MPO activities at 0 to 6 hours (p=0.001).
CONCLUSION
In the rodent model of 30 minutes occlusion and reperfusion, our study revealed, with using 99mTc-Tfc, that the TfR expression increased in the myocardium till 3 hours after reperfusion. TfR-mediated entry of iron into the cardiomyocytes may represent that this process plays a role in the I/R injury during the early reperfusion period.

Keyword

Reperfusion injury; Myocardium; Transferrin receptor

MeSH Terms

Animals
Autoradiography
Blotting, Western
Coronary Vessels
Eosine Yellowish-(YS)
Hematoxylin
Humans
Iron
Male
Myocardial Ischemia
Myocardium
Myocytes, Cardiac
Niacin
Peroxidase
Pilot Projects
Proteins
Rats
Receptors, Transferrin
Reperfusion
Reperfusion Injury
Rodentia
Transferrin
Eosine Yellowish-(YS)
Hematoxylin
Iron
Niacin
Peroxidase
Proteins
Receptors, Transferrin
Transferrin

Figure

• 

  Fig. 1 H & E staining according to the duration of reperfusion (×100). The neutrophil infiltration in the model of myocardial ischemia/reperfusion (I/R) became severe and then it became diffuse after the reperfused time went on for 24 hours. H & E: hematoxylin and eosin.

• 

  Fig. 2 Representative autoradiographic findings after injecting 37 MBq 99mTc transferrin-chitosan-HYNIC (99mTc-Tfc) via tail vein. The Tfc images demonstrate the area and intensity of binding to TfR. The 99mTc-Tfc uptake was increased with time after reperfusion until 3 hours and then it decreased at 3 hours. Thereafter, the uptake showed a diffuse pattern in the injured myocardial layer. Upper row: the auto-radiographic images, lower row: the sliced myocardial tissue for autoradiography. RT: reperfusion time, HYNIC: hydrazino nicotinate hydrochloride.

• 

  Fig. 3 Representative results of western blotting for transferrin receptor (TfR) and the myeloperoxidase (MPO) assay according to the duration of reperfusion time after occlusion. The amount of TfR proteins was increased at 0, 0.5 and 1.5 hours compared with that of the sham operation. The expression of TfR was changed from very low at 3 hours and it showed up again at 6 and 24 hours for a second time. The MPO activity only at 24 hours was significantly increased. *p<0.05. RT: reperfusion time.

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