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Dantrolene enhances the protective effect of hypothermia on cerebral cortex neurons.

Xu SY, Hu FY, Ren LJ, Chen L, Zhou ZQ, Zhang XJ, Li WP - Neural Regen Res (2015)

Bottom Line: Results revealed that the combination of dantrolene and hypothermia increased neuronal survival and the mitochondrial membrane potential, and reduced intracellular active oxygen cytoplasmic histone-associated DNA fragmentation, and apoptosis.Furthermore, improvements in cell morphology were observed.The combined treatment enhanced these responses compared with either treatment alone.

View Article: PubMed Central - PubMed

Affiliation: Postdoctoral Workstation, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong Province, China ; Department of Brain Center, Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People's Hospital, Shenzhen University First Affiliated Hospital, Shenzhen, Guangdong Province, China.

ABSTRACT
Therapeutic hypothermia is the most promising non-pharmacological neuroprotective strategy against ischemic injury. However, shivering is the most common adverse reaction. Many studies have shown that dantrolene is neuroprotective in in vitro and in vivo ischemic injury models. In addition to its neuroprotective effect, dantrolene neutralizes the adverse reaction of hypothermia. Dantrolene may be an effective adjunctive therapy to enhance the neuroprotection of hypothermia in treating ischemic stroke. Cortical neurons isolated from rat fetuses were exposed to 90 minutes of oxygen-glucose deprivation followed by reoxygenation. Neurons were treated with 40 μM dantrolene, hypothermia (at 33°C), or the combination of both for 12 hours. Results revealed that the combination of dantrolene and hypothermia increased neuronal survival and the mitochondrial membrane potential, and reduced intracellular active oxygen cytoplasmic histone-associated DNA fragmentation, and apoptosis. Furthermore, improvements in cell morphology were observed. The combined treatment enhanced these responses compared with either treatment alone. These findings indicate that dantrolene may be used as an effective adjunctive therapy to enhance the neuroprotective effects of hypothermia in ischemic stroke.

No MeSH data available.


Related in: MedlinePlus

Neuroprotection against OGD-mediated injury occurs with hypothermia, dantrolene, or their combined administration.DNA fragmentation and annexinV/PI labeling was performed 48 hours after OGD treatment to assess cell death, while other parameters were measured after 12 hours of hypothermia (or dantrolene) administration after OGD. (A) CCK-8: Cell viability. (B) DCFH-DA: ROS. (C) Fluo-3 AM: Free calcium ions. (D) JC-1: Monomers/J-aggregates. (E) Cell death detection: Cytoplasmic histone-associated DNA fragments. (F) Q1 + Q2: Late apoptosis and necrosis cells. Experiments were performed six times. *P < 0.05, vs. normal group; #P < 0.05, vs. OGD group; †P < 0.05, vs. H + D group. I: Normal group; II: OGD/R group; III: Hypothermia group; IV: Dantrolene group; IV: H + D. CCK-8: Cell counting kit-8; DCFH-DA: dichlorofluorescin diacetate; Fluo-3 AM: fluo-3-acetoxymethyl ester; OGD: oxygen-glucose deprivation; OGD/R: oxygen-glucose deprivation/reoxygenation; H + D: hypothermia and dantrolene; ROS: reactive oxygen species.
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Figure 5: Neuroprotection against OGD-mediated injury occurs with hypothermia, dantrolene, or their combined administration.DNA fragmentation and annexinV/PI labeling was performed 48 hours after OGD treatment to assess cell death, while other parameters were measured after 12 hours of hypothermia (or dantrolene) administration after OGD. (A) CCK-8: Cell viability. (B) DCFH-DA: ROS. (C) Fluo-3 AM: Free calcium ions. (D) JC-1: Monomers/J-aggregates. (E) Cell death detection: Cytoplasmic histone-associated DNA fragments. (F) Q1 + Q2: Late apoptosis and necrosis cells. Experiments were performed six times. *P < 0.05, vs. normal group; #P < 0.05, vs. OGD group; †P < 0.05, vs. H + D group. I: Normal group; II: OGD/R group; III: Hypothermia group; IV: Dantrolene group; IV: H + D. CCK-8: Cell counting kit-8; DCFH-DA: dichlorofluorescin diacetate; Fluo-3 AM: fluo-3-acetoxymethyl ester; OGD: oxygen-glucose deprivation; OGD/R: oxygen-glucose deprivation/reoxygenation; H + D: hypothermia and dantrolene; ROS: reactive oxygen species.

Mentions: OGD/R induced a dramatic (P < 0.05) reduction in cell viability compared to untreated controls (Figure 5A). This effect was markedly (P < 0.05) inhibited by hypothermia or dantrolene treatment, although this reduction was partial (Figure 5A). Furthermore, the combination of hypothermia and dantrolene was more effective in restoring cell viability than each treatment alone (P < 0.05).


Dantrolene enhances the protective effect of hypothermia on cerebral cortex neurons.

Xu SY, Hu FY, Ren LJ, Chen L, Zhou ZQ, Zhang XJ, Li WP - Neural Regen Res (2015)

Neuroprotection against OGD-mediated injury occurs with hypothermia, dantrolene, or their combined administration.DNA fragmentation and annexinV/PI labeling was performed 48 hours after OGD treatment to assess cell death, while other parameters were measured after 12 hours of hypothermia (or dantrolene) administration after OGD. (A) CCK-8: Cell viability. (B) DCFH-DA: ROS. (C) Fluo-3 AM: Free calcium ions. (D) JC-1: Monomers/J-aggregates. (E) Cell death detection: Cytoplasmic histone-associated DNA fragments. (F) Q1 + Q2: Late apoptosis and necrosis cells. Experiments were performed six times. *P < 0.05, vs. normal group; #P < 0.05, vs. OGD group; †P < 0.05, vs. H + D group. I: Normal group; II: OGD/R group; III: Hypothermia group; IV: Dantrolene group; IV: H + D. CCK-8: Cell counting kit-8; DCFH-DA: dichlorofluorescin diacetate; Fluo-3 AM: fluo-3-acetoxymethyl ester; OGD: oxygen-glucose deprivation; OGD/R: oxygen-glucose deprivation/reoxygenation; H + D: hypothermia and dantrolene; ROS: reactive oxygen species.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4590241&req=5

Figure 5: Neuroprotection against OGD-mediated injury occurs with hypothermia, dantrolene, or their combined administration.DNA fragmentation and annexinV/PI labeling was performed 48 hours after OGD treatment to assess cell death, while other parameters were measured after 12 hours of hypothermia (or dantrolene) administration after OGD. (A) CCK-8: Cell viability. (B) DCFH-DA: ROS. (C) Fluo-3 AM: Free calcium ions. (D) JC-1: Monomers/J-aggregates. (E) Cell death detection: Cytoplasmic histone-associated DNA fragments. (F) Q1 + Q2: Late apoptosis and necrosis cells. Experiments were performed six times. *P < 0.05, vs. normal group; #P < 0.05, vs. OGD group; †P < 0.05, vs. H + D group. I: Normal group; II: OGD/R group; III: Hypothermia group; IV: Dantrolene group; IV: H + D. CCK-8: Cell counting kit-8; DCFH-DA: dichlorofluorescin diacetate; Fluo-3 AM: fluo-3-acetoxymethyl ester; OGD: oxygen-glucose deprivation; OGD/R: oxygen-glucose deprivation/reoxygenation; H + D: hypothermia and dantrolene; ROS: reactive oxygen species.
Mentions: OGD/R induced a dramatic (P < 0.05) reduction in cell viability compared to untreated controls (Figure 5A). This effect was markedly (P < 0.05) inhibited by hypothermia or dantrolene treatment, although this reduction was partial (Figure 5A). Furthermore, the combination of hypothermia and dantrolene was more effective in restoring cell viability than each treatment alone (P < 0.05).

Bottom Line: Results revealed that the combination of dantrolene and hypothermia increased neuronal survival and the mitochondrial membrane potential, and reduced intracellular active oxygen cytoplasmic histone-associated DNA fragmentation, and apoptosis.Furthermore, improvements in cell morphology were observed.The combined treatment enhanced these responses compared with either treatment alone.

View Article: PubMed Central - PubMed

Affiliation: Postdoctoral Workstation, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong Province, China ; Department of Brain Center, Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People's Hospital, Shenzhen University First Affiliated Hospital, Shenzhen, Guangdong Province, China.

ABSTRACT
Therapeutic hypothermia is the most promising non-pharmacological neuroprotective strategy against ischemic injury. However, shivering is the most common adverse reaction. Many studies have shown that dantrolene is neuroprotective in in vitro and in vivo ischemic injury models. In addition to its neuroprotective effect, dantrolene neutralizes the adverse reaction of hypothermia. Dantrolene may be an effective adjunctive therapy to enhance the neuroprotection of hypothermia in treating ischemic stroke. Cortical neurons isolated from rat fetuses were exposed to 90 minutes of oxygen-glucose deprivation followed by reoxygenation. Neurons were treated with 40 μM dantrolene, hypothermia (at 33°C), or the combination of both for 12 hours. Results revealed that the combination of dantrolene and hypothermia increased neuronal survival and the mitochondrial membrane potential, and reduced intracellular active oxygen cytoplasmic histone-associated DNA fragmentation, and apoptosis. Furthermore, improvements in cell morphology were observed. The combined treatment enhanced these responses compared with either treatment alone. These findings indicate that dantrolene may be used as an effective adjunctive therapy to enhance the neuroprotective effects of hypothermia in ischemic stroke.

No MeSH data available.


Related in: MedlinePlus