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Neurosupportive Role of Vanillin, a Natural Phenolic Compound, on Rotenone Induced Neurotoxicity in SH-SY5Y Neuroblastoma Cells.

Dhanalakshmi C, Manivasagam T, Nataraj J, Justin Thenmozhi A, Essa MM - Evid Based Complement Alternat Med (2015)

Bottom Line: The therapeutic effectiveness of vanillin against rotenone was measured by pretreatment of vanillin at various concentrations (5-200 nM) and then incubation with rotenone (100 nM).Using effective dose of vanillin (100 nM), mitochondrial membrane potential, levels of reactive oxygen species (ROS), and expression patterns of apoptotic markers were assessed.Our results indicated that the pretreatment of vanillin attenuated rotenone induced mitochondrial dysfunction, oxidative stress, and apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Biotechnology, Annamalai University, Annamalainagar, Tamil Nadu 608002, India.

ABSTRACT
Vanillin, a phenolic compound, has been reported to offer neuroprotection against experimental Huntington's disease and global ischemia by virtue of its antioxidant, anti-inflammatory, and antiapoptotic properties. The present study aims to elucidate the underlying neuroprotective mechanism of vanillin in rotenone induced neurotoxicity. Cell viability was assessed by exposing SH-SY5Y cells to various concentrations of rotenone (5-200 nM) for 24 h. The therapeutic effectiveness of vanillin against rotenone was measured by pretreatment of vanillin at various concentrations (5-200 nM) and then incubation with rotenone (100 nM). Using effective dose of vanillin (100 nM), mitochondrial membrane potential, levels of reactive oxygen species (ROS), and expression patterns of apoptotic markers were assessed. Toxicity of rotenone was accompanied by the loss of mitochondrial membrane potential, increased ROS generation, release of cyt-c, and enhanced expressions of proapoptotic and downregulation of antiapoptotic indices via the upregulation of p38 and JNK-MAPK pathway proteins. Our results indicated that the pretreatment of vanillin attenuated rotenone induced mitochondrial dysfunction, oxidative stress, and apoptosis. Thus, vanillin may serve as a potent therapeutic agent in the future by virtue of its multiple pharmacological properties in the treatment of neurodegenerative diseases including PD.

No MeSH data available.


Related in: MedlinePlus

The effect of vanillin on the expressions of apoptotic and signaling markers. Lane 1: control; 2: rotenone; 3: vanillin + rotenone; and 4: vanillin. (a), (b), and (c) show the expressions of Bax; caspase-3, caspase-8, and caspase-9 cyt-c in cytosol were increased while the expressions of Bcl-2 and cyt-c in mitochondria were significantly decreased by the rotenone treated group as compared with control. Pretreatment with vanillin gradually restored the imbalanced expression profile of these proteins. (d) Rotenone treatment stimulates the expressions of p-JNK, p-P38, and p-ERK as compared with control. Pretreatment with vanillin decreases the expressions of p-JNK, p-P38, and p-ERK significantly. Immunoblots are representative of at least four independent experiments. Values are given as mean ± SD in each group. ∗p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).
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fig7: The effect of vanillin on the expressions of apoptotic and signaling markers. Lane 1: control; 2: rotenone; 3: vanillin + rotenone; and 4: vanillin. (a), (b), and (c) show the expressions of Bax; caspase-3, caspase-8, and caspase-9 cyt-c in cytosol were increased while the expressions of Bcl-2 and cyt-c in mitochondria were significantly decreased by the rotenone treated group as compared with control. Pretreatment with vanillin gradually restored the imbalanced expression profile of these proteins. (d) Rotenone treatment stimulates the expressions of p-JNK, p-P38, and p-ERK as compared with control. Pretreatment with vanillin decreases the expressions of p-JNK, p-P38, and p-ERK significantly. Immunoblots are representative of at least four independent experiments. Values are given as mean ± SD in each group. ∗p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).

Mentions: To further characterize the mechanism of inhibition by vanillin on rotenone induced apoptosis, we determined the effect of vanillin on the expression of anti- and proapoptotic proteins by western blot. The expression of Bax, caspase-3, caspase-8, and caspase-9 was increased while the distribution of Bcl-2 and cyt-c in mitochondria was significantly decreased by the rotenone treated group as compared with control. Rotenone treatment significantly diminished the translocation of cyt-c in cytosol. Pretreatment with vanillin gradually restored the imbalanced expression profile of these proteins. Vanillin, which had no effects in control cells, in contrast, when treated with rotenone noticeably, changes the protein expressions in SH-SY5Y cells (Figures 7(a), 7(b), and 7(c)). In order to elucidate the mechanism of rotenone induced cell death and protective effective of vanillin, the protein expression studies of signaling molecules were performed. Expressions of p-JNK, p-P38, and p-ERK were significantly increased after rotenone treatment as compared with control. Following pretreatment with vanillin before rotenone addition is able to decrease the levels of p-JNK, p-P38, and p-ERK significantly. However, treatment with vanillin alone did not alter the expression; p-JNK, p-P38, and p-ERK were unaffected as compared to control (Figure 7(d)).


Neurosupportive Role of Vanillin, a Natural Phenolic Compound, on Rotenone Induced Neurotoxicity in SH-SY5Y Neuroblastoma Cells.

Dhanalakshmi C, Manivasagam T, Nataraj J, Justin Thenmozhi A, Essa MM - Evid Based Complement Alternat Med (2015)

The effect of vanillin on the expressions of apoptotic and signaling markers. Lane 1: control; 2: rotenone; 3: vanillin + rotenone; and 4: vanillin. (a), (b), and (c) show the expressions of Bax; caspase-3, caspase-8, and caspase-9 cyt-c in cytosol were increased while the expressions of Bcl-2 and cyt-c in mitochondria were significantly decreased by the rotenone treated group as compared with control. Pretreatment with vanillin gradually restored the imbalanced expression profile of these proteins. (d) Rotenone treatment stimulates the expressions of p-JNK, p-P38, and p-ERK as compared with control. Pretreatment with vanillin decreases the expressions of p-JNK, p-P38, and p-ERK significantly. Immunoblots are representative of at least four independent experiments. Values are given as mean ± SD in each group. ∗p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4664805&req=5

fig7: The effect of vanillin on the expressions of apoptotic and signaling markers. Lane 1: control; 2: rotenone; 3: vanillin + rotenone; and 4: vanillin. (a), (b), and (c) show the expressions of Bax; caspase-3, caspase-8, and caspase-9 cyt-c in cytosol were increased while the expressions of Bcl-2 and cyt-c in mitochondria were significantly decreased by the rotenone treated group as compared with control. Pretreatment with vanillin gradually restored the imbalanced expression profile of these proteins. (d) Rotenone treatment stimulates the expressions of p-JNK, p-P38, and p-ERK as compared with control. Pretreatment with vanillin decreases the expressions of p-JNK, p-P38, and p-ERK significantly. Immunoblots are representative of at least four independent experiments. Values are given as mean ± SD in each group. ∗p < 0.05 compared to control and #p < 0.05 compared to rotenone group (DMRT).
Mentions: To further characterize the mechanism of inhibition by vanillin on rotenone induced apoptosis, we determined the effect of vanillin on the expression of anti- and proapoptotic proteins by western blot. The expression of Bax, caspase-3, caspase-8, and caspase-9 was increased while the distribution of Bcl-2 and cyt-c in mitochondria was significantly decreased by the rotenone treated group as compared with control. Rotenone treatment significantly diminished the translocation of cyt-c in cytosol. Pretreatment with vanillin gradually restored the imbalanced expression profile of these proteins. Vanillin, which had no effects in control cells, in contrast, when treated with rotenone noticeably, changes the protein expressions in SH-SY5Y cells (Figures 7(a), 7(b), and 7(c)). In order to elucidate the mechanism of rotenone induced cell death and protective effective of vanillin, the protein expression studies of signaling molecules were performed. Expressions of p-JNK, p-P38, and p-ERK were significantly increased after rotenone treatment as compared with control. Following pretreatment with vanillin before rotenone addition is able to decrease the levels of p-JNK, p-P38, and p-ERK significantly. However, treatment with vanillin alone did not alter the expression; p-JNK, p-P38, and p-ERK were unaffected as compared to control (Figure 7(d)).

Bottom Line: The therapeutic effectiveness of vanillin against rotenone was measured by pretreatment of vanillin at various concentrations (5-200 nM) and then incubation with rotenone (100 nM).Using effective dose of vanillin (100 nM), mitochondrial membrane potential, levels of reactive oxygen species (ROS), and expression patterns of apoptotic markers were assessed.Our results indicated that the pretreatment of vanillin attenuated rotenone induced mitochondrial dysfunction, oxidative stress, and apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Biotechnology, Annamalai University, Annamalainagar, Tamil Nadu 608002, India.

ABSTRACT
Vanillin, a phenolic compound, has been reported to offer neuroprotection against experimental Huntington's disease and global ischemia by virtue of its antioxidant, anti-inflammatory, and antiapoptotic properties. The present study aims to elucidate the underlying neuroprotective mechanism of vanillin in rotenone induced neurotoxicity. Cell viability was assessed by exposing SH-SY5Y cells to various concentrations of rotenone (5-200 nM) for 24 h. The therapeutic effectiveness of vanillin against rotenone was measured by pretreatment of vanillin at various concentrations (5-200 nM) and then incubation with rotenone (100 nM). Using effective dose of vanillin (100 nM), mitochondrial membrane potential, levels of reactive oxygen species (ROS), and expression patterns of apoptotic markers were assessed. Toxicity of rotenone was accompanied by the loss of mitochondrial membrane potential, increased ROS generation, release of cyt-c, and enhanced expressions of proapoptotic and downregulation of antiapoptotic indices via the upregulation of p38 and JNK-MAPK pathway proteins. Our results indicated that the pretreatment of vanillin attenuated rotenone induced mitochondrial dysfunction, oxidative stress, and apoptosis. Thus, vanillin may serve as a potent therapeutic agent in the future by virtue of its multiple pharmacological properties in the treatment of neurodegenerative diseases including PD.

No MeSH data available.


Related in: MedlinePlus