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Neuroprotective effects of ginsenoside Rb1 on high glucose-induced neurotoxicity in primary cultured rat hippocampal neurons.

Liu D, Zhang H, Gu W, Liu Y, Zhang M - PLoS ONE (2013)

Bottom Line: Ginsenoside Rb1 is one of the main active principles in traditional herb ginseng and has been reported to have a wide variety of neuroprotective effects.In addition, the high glucose-induced cell apoptosis, activation of ER stress, ROS accumulation and mitochondrial dysfunction can also be attenuated by the inhibitor of ER stress 4-phenylbutyric acid (4-PBA) and anti-oxidant N-acetylcysteine(NAC).In conclusion, these results suggest that Rb1 may protect neurons against high glucose-induced cell injury through inhibiting CHOP signaling pathway as well as oxidative stress and mitochondrial dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Department of Traditional Chinese Medicine, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.

ABSTRACT
Ginsenoside Rb1 is one of the main active principles in traditional herb ginseng and has been reported to have a wide variety of neuroprotective effects. Endoplasmic reticulum (ER) stress has been implicated in neurodegenerative diseases, so the present study aimed to observe the effects of ginsenoside Rb1 on ER stress signaling pathways in high glucose-treated hippocampal neurons. The results from MTT, TUNEL labeling and Annexin V-FITC/PI/Hoechst assays showed that incubating neurons with 50 mM high glucose for 72 h decreased cell viability and increased the number of apoptotic cells whereas treating neurons with 1 μM Rb1 for 72 h protected the neurons against high glucose-induced cell damage. Further molecular mechanism study demonstrated that Rb1 suppressed the activation of ER stress-associated proteins including protein kinase RNA (PKR)-like ER kinase (PERK) and C/EBP homology protein (CHOP) and downregulation of Bcl-2 induced by high glucose. Moreover, Rb1 inhibited both the elevation of intracellular reactive oxygen species (ROS) and the disruption of mitochondrial membrane potential induced by high glucose. In addition, the high glucose-induced cell apoptosis, activation of ER stress, ROS accumulation and mitochondrial dysfunction can also be attenuated by the inhibitor of ER stress 4-phenylbutyric acid (4-PBA) and anti-oxidant N-acetylcysteine(NAC). In conclusion, these results suggest that Rb1 may protect neurons against high glucose-induced cell injury through inhibiting CHOP signaling pathway as well as oxidative stress and mitochondrial dysfunction.

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Neuroprotective effects of ginsenoside Rb1 on cytotoxicity induced by high glucose.(A) Chemical structure of ginsenoside Rb1. (B) Primary hippocampal neurons were treated with ginsenoside Rb1(1μM) in the presence of 50 mM high glucose for 72h. Cell viability was measured using the MTT assay. The results represent the mean ± S.D of at least three independent experiments, and are expressed as percentage of control; **P<0.01, as compared to the control group; # P<0.05, as compared to the high glucose group ; ##P<0.01, as compared to the high glucose group.
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pone-0079399-g001: Neuroprotective effects of ginsenoside Rb1 on cytotoxicity induced by high glucose.(A) Chemical structure of ginsenoside Rb1. (B) Primary hippocampal neurons were treated with ginsenoside Rb1(1μM) in the presence of 50 mM high glucose for 72h. Cell viability was measured using the MTT assay. The results represent the mean ± S.D of at least three independent experiments, and are expressed as percentage of control; **P<0.01, as compared to the control group; # P<0.05, as compared to the high glucose group ; ##P<0.01, as compared to the high glucose group.

Mentions: Ginsenoside-Rb1 standard was obtained from National Institute for the Control of Pharmaceutical and Biological Produces (Beijing, China). The structure of ginsenoside Rb1 (2-O-β-Glucopyranosyl-(3β, 12β)-20-[(6-O-β -D-glucopyranosyl-β-D-glucopyranosyl) oxy]-12-hydroxydammar-24-en-3-yl β -D-glucopyranoside) is illustrated in Figure 1A. All cell culture reagents were from Peking union cell center (Beijing, China). In Situ Cell Death Detection Kit was purchased from Roche Diagnostics GmbH (Penzberg, Germany). Annexin V-FITC/PI detection kit, Hoechst 33258, fluorescent probe 2′, 7′-dichlorofluorescin diacetate (DCFH-DA) and 4-phenylbutyric acid (4-PBA) were obtained from Sigma-Aldrich (St. Loius, MO, USA). N-acetylcysteine(NAC) was purchased from TCI chemicals (Shanghai, China). Mitochondrial membrane potential assay kit with 5,5′,6,6′-tetrachloro- 1,1′ ,3,3′-tetraethyl benzimidazolcarbocyanine iodide (JC-1) was obtained from Beyotime Institute of Biotechnology(Shanghai, China). Anti-p-PERK, anti-PERK, anti-CHOP and anti-Bcl-2 antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).


Neuroprotective effects of ginsenoside Rb1 on high glucose-induced neurotoxicity in primary cultured rat hippocampal neurons.

Liu D, Zhang H, Gu W, Liu Y, Zhang M - PLoS ONE (2013)

Neuroprotective effects of ginsenoside Rb1 on cytotoxicity induced by high glucose.(A) Chemical structure of ginsenoside Rb1. (B) Primary hippocampal neurons were treated with ginsenoside Rb1(1μM) in the presence of 50 mM high glucose for 72h. Cell viability was measured using the MTT assay. The results represent the mean ± S.D of at least three independent experiments, and are expressed as percentage of control; **P<0.01, as compared to the control group; # P<0.05, as compared to the high glucose group ; ##P<0.01, as compared to the high glucose group.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3815219&req=5

pone-0079399-g001: Neuroprotective effects of ginsenoside Rb1 on cytotoxicity induced by high glucose.(A) Chemical structure of ginsenoside Rb1. (B) Primary hippocampal neurons were treated with ginsenoside Rb1(1μM) in the presence of 50 mM high glucose for 72h. Cell viability was measured using the MTT assay. The results represent the mean ± S.D of at least three independent experiments, and are expressed as percentage of control; **P<0.01, as compared to the control group; # P<0.05, as compared to the high glucose group ; ##P<0.01, as compared to the high glucose group.
Mentions: Ginsenoside-Rb1 standard was obtained from National Institute for the Control of Pharmaceutical and Biological Produces (Beijing, China). The structure of ginsenoside Rb1 (2-O-β-Glucopyranosyl-(3β, 12β)-20-[(6-O-β -D-glucopyranosyl-β-D-glucopyranosyl) oxy]-12-hydroxydammar-24-en-3-yl β -D-glucopyranoside) is illustrated in Figure 1A. All cell culture reagents were from Peking union cell center (Beijing, China). In Situ Cell Death Detection Kit was purchased from Roche Diagnostics GmbH (Penzberg, Germany). Annexin V-FITC/PI detection kit, Hoechst 33258, fluorescent probe 2′, 7′-dichlorofluorescin diacetate (DCFH-DA) and 4-phenylbutyric acid (4-PBA) were obtained from Sigma-Aldrich (St. Loius, MO, USA). N-acetylcysteine(NAC) was purchased from TCI chemicals (Shanghai, China). Mitochondrial membrane potential assay kit with 5,5′,6,6′-tetrachloro- 1,1′ ,3,3′-tetraethyl benzimidazolcarbocyanine iodide (JC-1) was obtained from Beyotime Institute of Biotechnology(Shanghai, China). Anti-p-PERK, anti-PERK, anti-CHOP and anti-Bcl-2 antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Bottom Line: Ginsenoside Rb1 is one of the main active principles in traditional herb ginseng and has been reported to have a wide variety of neuroprotective effects.In addition, the high glucose-induced cell apoptosis, activation of ER stress, ROS accumulation and mitochondrial dysfunction can also be attenuated by the inhibitor of ER stress 4-phenylbutyric acid (4-PBA) and anti-oxidant N-acetylcysteine(NAC).In conclusion, these results suggest that Rb1 may protect neurons against high glucose-induced cell injury through inhibiting CHOP signaling pathway as well as oxidative stress and mitochondrial dysfunction.

View Article: PubMed Central - PubMed

Affiliation: Department of Traditional Chinese Medicine, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China.

ABSTRACT
Ginsenoside Rb1 is one of the main active principles in traditional herb ginseng and has been reported to have a wide variety of neuroprotective effects. Endoplasmic reticulum (ER) stress has been implicated in neurodegenerative diseases, so the present study aimed to observe the effects of ginsenoside Rb1 on ER stress signaling pathways in high glucose-treated hippocampal neurons. The results from MTT, TUNEL labeling and Annexin V-FITC/PI/Hoechst assays showed that incubating neurons with 50 mM high glucose for 72 h decreased cell viability and increased the number of apoptotic cells whereas treating neurons with 1 μM Rb1 for 72 h protected the neurons against high glucose-induced cell damage. Further molecular mechanism study demonstrated that Rb1 suppressed the activation of ER stress-associated proteins including protein kinase RNA (PKR)-like ER kinase (PERK) and C/EBP homology protein (CHOP) and downregulation of Bcl-2 induced by high glucose. Moreover, Rb1 inhibited both the elevation of intracellular reactive oxygen species (ROS) and the disruption of mitochondrial membrane potential induced by high glucose. In addition, the high glucose-induced cell apoptosis, activation of ER stress, ROS accumulation and mitochondrial dysfunction can also be attenuated by the inhibitor of ER stress 4-phenylbutyric acid (4-PBA) and anti-oxidant N-acetylcysteine(NAC). In conclusion, these results suggest that Rb1 may protect neurons against high glucose-induced cell injury through inhibiting CHOP signaling pathway as well as oxidative stress and mitochondrial dysfunction.

Show MeSH
Related in: MedlinePlus