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Combination treatment with ethyl pyruvate and IGF-I exerts neuroprotective effects against brain injury in a rat model of neonatal hypoxic-ischemic encephalopathy.

Rong Z, Pan R, Chang L, Lee W - Int. J. Mol. Med. (2015)

Bottom Line: Under conditions of OGD, the LDH levels increased and neuronal viability decreased.Treatment with 0.5 mM EP or 25 ng/ml IGF‑I protected the neurons (P<0.05), exerting additive effects.This effect may be the result of reduced neuronal injury, and enhanced neurogenesis and maturation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.

ABSTRACT
Neonatal hypoxic-ischemic (HI) brain injury causes severe brain damage in newborns. Following HI injury, rapidly accumulating oxidants injure neurons and interrupt ongoing developmental processes. The antioxidant, sodium pyruvate, has been shown to reduce neuronal injury in neonatal rats under conditions of oxygen glucose deprivation (OGD) and HI injury. In this study, we evaluated the effects of ethyl pyruvate (EP) and insulin‑like growth factor‑I (IGF‑I) alone or in combination in a similar setting. For this purpose, we used an in vitro model involving primary neonatal rat cortical neurons subjected to OGD for 2.5 h and an in vivo model involving unilateral carotid ligation in rats on post-natal day 7 with exposure to 8% hypoxia for 2.5 h. The cultured neurons were examined by lactate dehydrogenase (LDH) and cell viability assays. For the in vivo experiments, behavioral development was evaluated by the foot fault test at 4 weeks of recovery. 2,3,5‑Triphenyltetrazolium chloride monohydrate and cresyl violet staining were used to evaluate HI injury. The injured neurons were Fluoro‑Jade B-labeled, new neuroprecursors were double labeled with bromodeoxyuridine (BrdU) and doublecortin, new mature neurons were BrdU-labeled and neuronal nuclei were labeled by immunofluorescence. Under conditions of OGD, the LDH levels increased and neuronal viability decreased. Treatment with 0.5 mM EP or 25 ng/ml IGF‑I protected the neurons (P<0.05), exerting additive effects. Similarly, either the early administration of EP or delayed treatment with IGF‑I protected the neonatal rat brains against HI injury and improved neurological performance and these effects were also additive. This effect may be the result of reduced neuronal injury, and enhanced neurogenesis and maturation. On the whole, our findings demonstrate that the combination of the early administration of EP with delayed treatment with IGF‑I exerts neuroprotective effects against HI injury in neonatal rat brains.

No MeSH data available.


Related in: MedlinePlus

Combined treatment with ethyl pyruvate (EP) and insulin-like growth factor-I (IGF-I) decreases hypoxic-ischemic (HI) injury-induced neuronal cell death in the hippocampal cornu ammonis 3 (CA3) region. (A) At 72 h post-HI injury, the injured hippocampal neurons were identified by Fluoro-Jade B (FJB) staining (green) and the nucleus were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). (B) High magnification view [squares in (A)] showing the morphology of degenerating hippocampal neurons (green). (C) Cell nuclei were identified by DAPI staining. (D) Merged image of (B) and (C). (E) Quantification of FJB+ cells in the ipsilateral hippocampus. *P<0.05 as indicated (n=3–6 per group).
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f4-ijmm-36-01-0195: Combined treatment with ethyl pyruvate (EP) and insulin-like growth factor-I (IGF-I) decreases hypoxic-ischemic (HI) injury-induced neuronal cell death in the hippocampal cornu ammonis 3 (CA3) region. (A) At 72 h post-HI injury, the injured hippocampal neurons were identified by Fluoro-Jade B (FJB) staining (green) and the nucleus were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). (B) High magnification view [squares in (A)] showing the morphology of degenerating hippocampal neurons (green). (C) Cell nuclei were identified by DAPI staining. (D) Merged image of (B) and (C). (E) Quantification of FJB+ cells in the ipsilateral hippocampus. *P<0.05 as indicated (n=3–6 per group).

Mentions: The increase in the volume of surviving tissue in the brain may have been a result of a reduced number of injured neurons. To clarify this, we labeled the injured neurons with FJB beginning at 3 h after injury and for up to at least 7 days. Fig. 4 illustrates the distribution of FJB+ cells in the hippocampus. The FJB+ neurons were detected at 3 h of recovery and reached peak numbers from 48–72 h (data not shown). At 72 h of recovery, HI injury to the neonatal brain increased the number of FJB+ cells both in the dentate gyrus (DG) and cornu ammonis 3 (CA3) region of the hippocampus (Fig. 4A–D). At a higher magnification, the FJB+ neurons displayed distinct apoptotic nuclei with either a condensed or fragmented morphology. Compared with the vehicle-treated group, the numbers of FJB+ neurons were significantly decreased by EP treatment or IGF-I treatment, whereas combined treatment with EP and IGF-I led to a further decrease in the number of FJB+ neurons compared to the groups treated with EP or IGF-I alone (Fig. 4E).


Combination treatment with ethyl pyruvate and IGF-I exerts neuroprotective effects against brain injury in a rat model of neonatal hypoxic-ischemic encephalopathy.

Rong Z, Pan R, Chang L, Lee W - Int. J. Mol. Med. (2015)

Combined treatment with ethyl pyruvate (EP) and insulin-like growth factor-I (IGF-I) decreases hypoxic-ischemic (HI) injury-induced neuronal cell death in the hippocampal cornu ammonis 3 (CA3) region. (A) At 72 h post-HI injury, the injured hippocampal neurons were identified by Fluoro-Jade B (FJB) staining (green) and the nucleus were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). (B) High magnification view [squares in (A)] showing the morphology of degenerating hippocampal neurons (green). (C) Cell nuclei were identified by DAPI staining. (D) Merged image of (B) and (C). (E) Quantification of FJB+ cells in the ipsilateral hippocampus. *P<0.05 as indicated (n=3–6 per group).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494588&req=5

f4-ijmm-36-01-0195: Combined treatment with ethyl pyruvate (EP) and insulin-like growth factor-I (IGF-I) decreases hypoxic-ischemic (HI) injury-induced neuronal cell death in the hippocampal cornu ammonis 3 (CA3) region. (A) At 72 h post-HI injury, the injured hippocampal neurons were identified by Fluoro-Jade B (FJB) staining (green) and the nucleus were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). (B) High magnification view [squares in (A)] showing the morphology of degenerating hippocampal neurons (green). (C) Cell nuclei were identified by DAPI staining. (D) Merged image of (B) and (C). (E) Quantification of FJB+ cells in the ipsilateral hippocampus. *P<0.05 as indicated (n=3–6 per group).
Mentions: The increase in the volume of surviving tissue in the brain may have been a result of a reduced number of injured neurons. To clarify this, we labeled the injured neurons with FJB beginning at 3 h after injury and for up to at least 7 days. Fig. 4 illustrates the distribution of FJB+ cells in the hippocampus. The FJB+ neurons were detected at 3 h of recovery and reached peak numbers from 48–72 h (data not shown). At 72 h of recovery, HI injury to the neonatal brain increased the number of FJB+ cells both in the dentate gyrus (DG) and cornu ammonis 3 (CA3) region of the hippocampus (Fig. 4A–D). At a higher magnification, the FJB+ neurons displayed distinct apoptotic nuclei with either a condensed or fragmented morphology. Compared with the vehicle-treated group, the numbers of FJB+ neurons were significantly decreased by EP treatment or IGF-I treatment, whereas combined treatment with EP and IGF-I led to a further decrease in the number of FJB+ neurons compared to the groups treated with EP or IGF-I alone (Fig. 4E).

Bottom Line: Under conditions of OGD, the LDH levels increased and neuronal viability decreased.Treatment with 0.5 mM EP or 25 ng/ml IGF‑I protected the neurons (P<0.05), exerting additive effects.This effect may be the result of reduced neuronal injury, and enhanced neurogenesis and maturation.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China.

ABSTRACT
Neonatal hypoxic-ischemic (HI) brain injury causes severe brain damage in newborns. Following HI injury, rapidly accumulating oxidants injure neurons and interrupt ongoing developmental processes. The antioxidant, sodium pyruvate, has been shown to reduce neuronal injury in neonatal rats under conditions of oxygen glucose deprivation (OGD) and HI injury. In this study, we evaluated the effects of ethyl pyruvate (EP) and insulin‑like growth factor‑I (IGF‑I) alone or in combination in a similar setting. For this purpose, we used an in vitro model involving primary neonatal rat cortical neurons subjected to OGD for 2.5 h and an in vivo model involving unilateral carotid ligation in rats on post-natal day 7 with exposure to 8% hypoxia for 2.5 h. The cultured neurons were examined by lactate dehydrogenase (LDH) and cell viability assays. For the in vivo experiments, behavioral development was evaluated by the foot fault test at 4 weeks of recovery. 2,3,5‑Triphenyltetrazolium chloride monohydrate and cresyl violet staining were used to evaluate HI injury. The injured neurons were Fluoro‑Jade B-labeled, new neuroprecursors were double labeled with bromodeoxyuridine (BrdU) and doublecortin, new mature neurons were BrdU-labeled and neuronal nuclei were labeled by immunofluorescence. Under conditions of OGD, the LDH levels increased and neuronal viability decreased. Treatment with 0.5 mM EP or 25 ng/ml IGF‑I protected the neurons (P<0.05), exerting additive effects. Similarly, either the early administration of EP or delayed treatment with IGF‑I protected the neonatal rat brains against HI injury and improved neurological performance and these effects were also additive. This effect may be the result of reduced neuronal injury, and enhanced neurogenesis and maturation. On the whole, our findings demonstrate that the combination of the early administration of EP with delayed treatment with IGF‑I exerts neuroprotective effects against HI injury in neonatal rat brains.

No MeSH data available.


Related in: MedlinePlus