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BDNF-TrkB pathway mediates neuroprotection of hydrogen sulfide against formaldehyde-induced toxicity to PC12 cells.

Jiang JM, Zhou CF, Gao SL, Tian Y, Wang CY, Wang L, Gu HF, Tang XQ - PLoS ONE (2015)

Bottom Line: In the present study, we found that NaHS, a donor of H2S, upregulated the level of BDNF protein in PC12 cells, and significantly rescued FA-induced downregulation of BDNF levels.Furthermore, we found that pretreatment of PC12 cells with K252a, an inhibitor of the BDNF receptor TrkB, markedly reversed the inhibition of NaHS on FA-induced cytotoxicity and ablated the protective effects of NaHS on FA-induced oxidative stress, including the accumulation of intracellular reactive oxygen species (ROS), 4-hydroxy-2-trans-nonenal (4-HNE), and malondialdehyde (MDA).We also showed that K252a abolished the inhibition of NaHS on FA-induced apoptosis, as well as the activation of caspase-3 in PC12 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology & Institute of Neuroscience, Medical College, University of South China, Hengyang, 42100, Hunan, P. R. China; Key Laboratory for Cognitive Disorders and Neurodegenerative Diseases, University of South China, Hengyang, 421001, Hunan, P. R. China.

ABSTRACT
Formaldehyde (FA) is a common environmental contaminant that has toxic effects on the central nervous system (CNS). Our previous data demonstrated that hydrogen sulfide (H2S), the third endogenous gaseous mediator, has protective effects against FA-induced neurotoxicity. As is known to all, Brain-derived neurotropic factor (BDNF), a member of the neurotrophin gene family, mediates its neuroprotective properties via various intracellular signaling pathways triggered by activating the tyrosine kinase receptor B (TrkB). Intriguingly, our previous data have illustrated the upregulatory role of H2S on BDNF protein expression in the hippocampus of rats. Therefore, in this study, we hypothesized that H2S provides neuroprotection against FA toxicity by regulating BDNF-TrkB pathway. In the present study, we found that NaHS, a donor of H2S, upregulated the level of BDNF protein in PC12 cells, and significantly rescued FA-induced downregulation of BDNF levels. Furthermore, we found that pretreatment of PC12 cells with K252a, an inhibitor of the BDNF receptor TrkB, markedly reversed the inhibition of NaHS on FA-induced cytotoxicity and ablated the protective effects of NaHS on FA-induced oxidative stress, including the accumulation of intracellular reactive oxygen species (ROS), 4-hydroxy-2-trans-nonenal (4-HNE), and malondialdehyde (MDA). We also showed that K252a abolished the inhibition of NaHS on FA-induced apoptosis, as well as the activation of caspase-3 in PC12 cells. In addition, K252a reversed the protection of H2S against FA-induced downregulation of Bcl-2 protein expression and upregulation of Bax protein expression in PC12 cells. These data indicate that the BDNF-TrkB pathway mediates the neuroprotection of H2S against FA-induced cytotoxicity, oxidative stress and apoptosis in PC12 cells. These findings provide a novel mechanism underlying the protection of H2S against FA-induced neurotoxicity.

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Effect of K252a on H2S-caused suppression in formaldehyde-induced upregulation of Bax and downregulation of Bcl-2 in PC12 cells.PC12 cells were pre-incubated with K252a (10 nM) for 30 min before pretreatment with NaHS (200 μM) for 30 min prior and the cotreated with formaldehyde (FA, 120 μM) for 24 h. The expression of Bax (A) and Bcl-2 (B) were detected by Western blot using anti-Bax antibody and anti-Bcl-2 antibody, respectively. In all blots, β-actin was used as a loading control. The ratio of Bax or Bcl-2 to β-actin was normalized by the value in control group. Values were expressed as the mean ± S.E.M. of three independent experiments. **P < 0.01, versus control group; ##P <0.01, versus NaHS-treated alone group; $ $P <0.01, versus FA-treated alone group; &&P < 0.01, versus cotreated with NaHS and FA group.
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pone.0119478.g006: Effect of K252a on H2S-caused suppression in formaldehyde-induced upregulation of Bax and downregulation of Bcl-2 in PC12 cells.PC12 cells were pre-incubated with K252a (10 nM) for 30 min before pretreatment with NaHS (200 μM) for 30 min prior and the cotreated with formaldehyde (FA, 120 μM) for 24 h. The expression of Bax (A) and Bcl-2 (B) were detected by Western blot using anti-Bax antibody and anti-Bcl-2 antibody, respectively. In all blots, β-actin was used as a loading control. The ratio of Bax or Bcl-2 to β-actin was normalized by the value in control group. Values were expressed as the mean ± S.E.M. of three independent experiments. **P < 0.01, versus control group; ##P <0.01, versus NaHS-treated alone group; $ $P <0.01, versus FA-treated alone group; &&P < 0.01, versus cotreated with NaHS and FA group.

Mentions: Finally, we investigated whether BDNF-TrkB pathway mediates the protective effect of H2S against FA-induced change in apoptosis-related proteins in PC12 cells. We found that pretreatment of PC12 cells with K252a (10 nM, for 30 min) reverses the protection of NaHS against FA-induced upregulation of Bax protein expression (Fig. 6A) and downregulation of Bcl-2 protein expression (Fig. 6B). Notably, treatment with NaHS alone (200 μM, 24 h) decreased the levels of Bax (Fig. 6A) and increased the levels of Bcl-2 (Fig. 6B) in PC12 cells. However, co-treatment with K252a (10 nM) and NaHS(200 μM) for 24 h significantly abolished the NaHS-induced downregulation of Bax (Fig. 6A) and upregulation of Bcl-2 (Fig. 6B). These results indicated that BDNF-TrkB pathway is able to mediate the inhibitory role of H2S in FA-induced proapoptotic potential.


BDNF-TrkB pathway mediates neuroprotection of hydrogen sulfide against formaldehyde-induced toxicity to PC12 cells.

Jiang JM, Zhou CF, Gao SL, Tian Y, Wang CY, Wang L, Gu HF, Tang XQ - PLoS ONE (2015)

Effect of K252a on H2S-caused suppression in formaldehyde-induced upregulation of Bax and downregulation of Bcl-2 in PC12 cells.PC12 cells were pre-incubated with K252a (10 nM) for 30 min before pretreatment with NaHS (200 μM) for 30 min prior and the cotreated with formaldehyde (FA, 120 μM) for 24 h. The expression of Bax (A) and Bcl-2 (B) were detected by Western blot using anti-Bax antibody and anti-Bcl-2 antibody, respectively. In all blots, β-actin was used as a loading control. The ratio of Bax or Bcl-2 to β-actin was normalized by the value in control group. Values were expressed as the mean ± S.E.M. of three independent experiments. **P < 0.01, versus control group; ##P <0.01, versus NaHS-treated alone group; $ $P <0.01, versus FA-treated alone group; &&P < 0.01, versus cotreated with NaHS and FA group.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4352058&req=5

pone.0119478.g006: Effect of K252a on H2S-caused suppression in formaldehyde-induced upregulation of Bax and downregulation of Bcl-2 in PC12 cells.PC12 cells were pre-incubated with K252a (10 nM) for 30 min before pretreatment with NaHS (200 μM) for 30 min prior and the cotreated with formaldehyde (FA, 120 μM) for 24 h. The expression of Bax (A) and Bcl-2 (B) were detected by Western blot using anti-Bax antibody and anti-Bcl-2 antibody, respectively. In all blots, β-actin was used as a loading control. The ratio of Bax or Bcl-2 to β-actin was normalized by the value in control group. Values were expressed as the mean ± S.E.M. of three independent experiments. **P < 0.01, versus control group; ##P <0.01, versus NaHS-treated alone group; $ $P <0.01, versus FA-treated alone group; &&P < 0.01, versus cotreated with NaHS and FA group.
Mentions: Finally, we investigated whether BDNF-TrkB pathway mediates the protective effect of H2S against FA-induced change in apoptosis-related proteins in PC12 cells. We found that pretreatment of PC12 cells with K252a (10 nM, for 30 min) reverses the protection of NaHS against FA-induced upregulation of Bax protein expression (Fig. 6A) and downregulation of Bcl-2 protein expression (Fig. 6B). Notably, treatment with NaHS alone (200 μM, 24 h) decreased the levels of Bax (Fig. 6A) and increased the levels of Bcl-2 (Fig. 6B) in PC12 cells. However, co-treatment with K252a (10 nM) and NaHS(200 μM) for 24 h significantly abolished the NaHS-induced downregulation of Bax (Fig. 6A) and upregulation of Bcl-2 (Fig. 6B). These results indicated that BDNF-TrkB pathway is able to mediate the inhibitory role of H2S in FA-induced proapoptotic potential.

Bottom Line: In the present study, we found that NaHS, a donor of H2S, upregulated the level of BDNF protein in PC12 cells, and significantly rescued FA-induced downregulation of BDNF levels.Furthermore, we found that pretreatment of PC12 cells with K252a, an inhibitor of the BDNF receptor TrkB, markedly reversed the inhibition of NaHS on FA-induced cytotoxicity and ablated the protective effects of NaHS on FA-induced oxidative stress, including the accumulation of intracellular reactive oxygen species (ROS), 4-hydroxy-2-trans-nonenal (4-HNE), and malondialdehyde (MDA).We also showed that K252a abolished the inhibition of NaHS on FA-induced apoptosis, as well as the activation of caspase-3 in PC12 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology & Institute of Neuroscience, Medical College, University of South China, Hengyang, 42100, Hunan, P. R. China; Key Laboratory for Cognitive Disorders and Neurodegenerative Diseases, University of South China, Hengyang, 421001, Hunan, P. R. China.

ABSTRACT
Formaldehyde (FA) is a common environmental contaminant that has toxic effects on the central nervous system (CNS). Our previous data demonstrated that hydrogen sulfide (H2S), the third endogenous gaseous mediator, has protective effects against FA-induced neurotoxicity. As is known to all, Brain-derived neurotropic factor (BDNF), a member of the neurotrophin gene family, mediates its neuroprotective properties via various intracellular signaling pathways triggered by activating the tyrosine kinase receptor B (TrkB). Intriguingly, our previous data have illustrated the upregulatory role of H2S on BDNF protein expression in the hippocampus of rats. Therefore, in this study, we hypothesized that H2S provides neuroprotection against FA toxicity by regulating BDNF-TrkB pathway. In the present study, we found that NaHS, a donor of H2S, upregulated the level of BDNF protein in PC12 cells, and significantly rescued FA-induced downregulation of BDNF levels. Furthermore, we found that pretreatment of PC12 cells with K252a, an inhibitor of the BDNF receptor TrkB, markedly reversed the inhibition of NaHS on FA-induced cytotoxicity and ablated the protective effects of NaHS on FA-induced oxidative stress, including the accumulation of intracellular reactive oxygen species (ROS), 4-hydroxy-2-trans-nonenal (4-HNE), and malondialdehyde (MDA). We also showed that K252a abolished the inhibition of NaHS on FA-induced apoptosis, as well as the activation of caspase-3 in PC12 cells. In addition, K252a reversed the protection of H2S against FA-induced downregulation of Bcl-2 protein expression and upregulation of Bax protein expression in PC12 cells. These data indicate that the BDNF-TrkB pathway mediates the neuroprotection of H2S against FA-induced cytotoxicity, oxidative stress and apoptosis in PC12 cells. These findings provide a novel mechanism underlying the protection of H2S against FA-induced neurotoxicity.

Show MeSH
Related in: MedlinePlus