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Baicalein antagonizes rotenone-induced apoptosis in dopaminergic SH-SY5Y cells related to Parkinsonism.

Song JX, Choi MY, Wong KC, Chung WW, Sze SC, Ng TB, Zhang KY - Chin Med (2012)

Bottom Line: Baicalein significantly increased viability and decreased rotenone-induced death of SH-SY5Y cells in a dose-dependent manner.Baicalein antagonized rotenone-induced overproduction of ROS, loss of ΔΨm, the increased expression of Bax, cleaved caspase-3 and phosphorylated ERK1/2 and the decreased expression of Bcl-2.The antioxidative effect, mitochondrial protection and modulation of anti-and pro-apoptotic proteins are related to the neuroprotective effects of baicalein against rotenone induced cell death in SH-SY5Y cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Chinese Medicine, The University of Hong Kong, 10 Sassoon Road, Pokfulam, Hong Kong SAR, China. zhang.yanbo@yahoo.com.

ABSTRACT

Background: Two active compounds, baicalein and its glycoside baicalin were found in the dried root of Scutellaria baicalensis Georgi, and reported to be neuroprotective in vitro and in vivo. This study aims to evaluate the protective effects of baicalein on the rotenone-induced apoptosis in dopaminergic SH-SY5Y cells related to parkinsonism.

Methods: Cell viability and cytotoxicity were determined by MTT assay. The degree of nuclear apoptosis was evaluated with a fluorescent DNA-binding probe Hoechst 33258. The production of reactive oxidative species (ROS) and loss of mitochondrial membrane potential (ΔΨm) were determined by fluorescent staining with DCFH-DA and Rhodanmine 123, respectively. The expression of Bax, Bcl-2, cleaved caspase-3 and phosphorylated ERK1/2 was determined by the Western blots.

Results: Baicalein significantly increased viability and decreased rotenone-induced death of SH-SY5Y cells in a dose-dependent manner. Pre- and subsequent co-treatment with baicalein preserved the cell morphology and attenuated the nuclear apoptotic characteristics triggered by rotenone. Baicalein antagonized rotenone-induced overproduction of ROS, loss of ΔΨm, the increased expression of Bax, cleaved caspase-3 and phosphorylated ERK1/2 and the decreased expression of Bcl-2.

Conclusion: The antioxidative effect, mitochondrial protection and modulation of anti-and pro-apoptotic proteins are related to the neuroprotective effects of baicalein against rotenone induced cell death in SH-SY5Y cells.

No MeSH data available.


Related in: MedlinePlus

Effects of baicalein (Bai) on rotenone (RT)-induced imbalance in the expression of Bax, Bcl-2, cleaved caspase-3 and phopho-ERK1/2. Cells were pretreated with Bai for 1 hour and then cotreated with 20 μM RT for 24 hours in serum-free medium. Blots were stripped and reprobed for β-actin as a loading control. (A) Representative protein bands. (B) Statistical analysis. The corresponding bar graph represented data quantified from three independent experiments (n = 3, #P < 0.05 versus control, *P < 0.05 versus RT treatment, **P < 0.05 versus control).
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Figure 6: Effects of baicalein (Bai) on rotenone (RT)-induced imbalance in the expression of Bax, Bcl-2, cleaved caspase-3 and phopho-ERK1/2. Cells were pretreated with Bai for 1 hour and then cotreated with 20 μM RT for 24 hours in serum-free medium. Blots were stripped and reprobed for β-actin as a loading control. (A) Representative protein bands. (B) Statistical analysis. The corresponding bar graph represented data quantified from three independent experiments (n = 3, #P < 0.05 versus control, *P < 0.05 versus RT treatment, **P < 0.05 versus control).

Mentions: To further characterize the mechanism of baicalein inhibition on rotenone-induced apoptosis, we determined the effect of baicalein on the expression of anti- and pro-apoptotic proteins by Western blots. As shown in Figure 6, the expression of Bax and cleaved caspase-3 was increased while the expression of Bcl-2 was significantly decreased by the treatment with rotenone (20 μM) for 24 hours (P < 0.05), compared with the control. Pre- and subsequent co-treatment with increasing concentrations of baicalein gradually restored the imbalanced expression profile of these proteins. Interestingly, baicalein treatment alone for 24 hours could reduce the base levels of Bax (0.86 ± 0.07) and cleaved caspase-3 (0.71 ± 0.09) (P < 0.05).


Baicalein antagonizes rotenone-induced apoptosis in dopaminergic SH-SY5Y cells related to Parkinsonism.

Song JX, Choi MY, Wong KC, Chung WW, Sze SC, Ng TB, Zhang KY - Chin Med (2012)

Effects of baicalein (Bai) on rotenone (RT)-induced imbalance in the expression of Bax, Bcl-2, cleaved caspase-3 and phopho-ERK1/2. Cells were pretreated with Bai for 1 hour and then cotreated with 20 μM RT for 24 hours in serum-free medium. Blots were stripped and reprobed for β-actin as a loading control. (A) Representative protein bands. (B) Statistical analysis. The corresponding bar graph represented data quantified from three independent experiments (n = 3, #P < 0.05 versus control, *P < 0.05 versus RT treatment, **P < 0.05 versus control).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3275529&req=5

Figure 6: Effects of baicalein (Bai) on rotenone (RT)-induced imbalance in the expression of Bax, Bcl-2, cleaved caspase-3 and phopho-ERK1/2. Cells were pretreated with Bai for 1 hour and then cotreated with 20 μM RT for 24 hours in serum-free medium. Blots were stripped and reprobed for β-actin as a loading control. (A) Representative protein bands. (B) Statistical analysis. The corresponding bar graph represented data quantified from three independent experiments (n = 3, #P < 0.05 versus control, *P < 0.05 versus RT treatment, **P < 0.05 versus control).
Mentions: To further characterize the mechanism of baicalein inhibition on rotenone-induced apoptosis, we determined the effect of baicalein on the expression of anti- and pro-apoptotic proteins by Western blots. As shown in Figure 6, the expression of Bax and cleaved caspase-3 was increased while the expression of Bcl-2 was significantly decreased by the treatment with rotenone (20 μM) for 24 hours (P < 0.05), compared with the control. Pre- and subsequent co-treatment with increasing concentrations of baicalein gradually restored the imbalanced expression profile of these proteins. Interestingly, baicalein treatment alone for 24 hours could reduce the base levels of Bax (0.86 ± 0.07) and cleaved caspase-3 (0.71 ± 0.09) (P < 0.05).

Bottom Line: Baicalein significantly increased viability and decreased rotenone-induced death of SH-SY5Y cells in a dose-dependent manner.Baicalein antagonized rotenone-induced overproduction of ROS, loss of ΔΨm, the increased expression of Bax, cleaved caspase-3 and phosphorylated ERK1/2 and the decreased expression of Bcl-2.The antioxidative effect, mitochondrial protection and modulation of anti-and pro-apoptotic proteins are related to the neuroprotective effects of baicalein against rotenone induced cell death in SH-SY5Y cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: School of Chinese Medicine, The University of Hong Kong, 10 Sassoon Road, Pokfulam, Hong Kong SAR, China. zhang.yanbo@yahoo.com.

ABSTRACT

Background: Two active compounds, baicalein and its glycoside baicalin were found in the dried root of Scutellaria baicalensis Georgi, and reported to be neuroprotective in vitro and in vivo. This study aims to evaluate the protective effects of baicalein on the rotenone-induced apoptosis in dopaminergic SH-SY5Y cells related to parkinsonism.

Methods: Cell viability and cytotoxicity were determined by MTT assay. The degree of nuclear apoptosis was evaluated with a fluorescent DNA-binding probe Hoechst 33258. The production of reactive oxidative species (ROS) and loss of mitochondrial membrane potential (ΔΨm) were determined by fluorescent staining with DCFH-DA and Rhodanmine 123, respectively. The expression of Bax, Bcl-2, cleaved caspase-3 and phosphorylated ERK1/2 was determined by the Western blots.

Results: Baicalein significantly increased viability and decreased rotenone-induced death of SH-SY5Y cells in a dose-dependent manner. Pre- and subsequent co-treatment with baicalein preserved the cell morphology and attenuated the nuclear apoptotic characteristics triggered by rotenone. Baicalein antagonized rotenone-induced overproduction of ROS, loss of ΔΨm, the increased expression of Bax, cleaved caspase-3 and phosphorylated ERK1/2 and the decreased expression of Bcl-2.

Conclusion: The antioxidative effect, mitochondrial protection and modulation of anti-and pro-apoptotic proteins are related to the neuroprotective effects of baicalein against rotenone induced cell death in SH-SY5Y cells.

No MeSH data available.


Related in: MedlinePlus