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Schisandrin B inhibits cell growth and induces cellular apoptosis and autophagy in mouse hepatocytes and macrophages: implications for its hepatotoxicity.

Zhang Y, Zhou ZW, Jin H, Hu C, He ZX, Yu ZL, Ko KM, Yang T, Zhang X, Pan SY, Zhou SF - Drug Des Devel Ther (2015)

Bottom Line: However, the potential hepatotoxicity of Sch B is a major safety concern, and the underlying mechanism for Sch B-induced liver toxic effects is not fully elucidated.The results have shown that Sch B exhibits potent grow inhibitory, proapoptotic, and proautophagic effects in AML-12 and RAW 264.7 cells.More studies are warranted to fully delineate the underlying mechanisms, efficacy, and safety of Sch B for clinical use.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, People's Republic of China ; Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.

ABSTRACT
A number of drugs and herbal compounds have been documented to cause hepatoxicity. Schisandrin B (Sch B) is an active dibenzocyclooctadiene isolated from Schisandrae fructus, with a wide array of pharmacological activities. However, the potential hepatotoxicity of Sch B is a major safety concern, and the underlying mechanism for Sch B-induced liver toxic effects is not fully elucidated. In the present study, we aimed to investigate the liver toxic effects and the molecular mechanisms of Sch B in mouse liver and macrophage cells. The results have shown that Sch B exhibits potent grow inhibitory, proapoptotic, and proautophagic effects in AML-12 and RAW 264.7 cells. Sch B markedly arrested cells in G1 phase in both cell lines, accompanied by the down-regulation of cyclin dependent kinase 2 (CDK2) and cyclin D1 and up-regulation of p27 Kip1 and checkpoint kinase 1. Furthermore, Sch B markedly increased the apoptosis of AML-12 and RAW 264.7 cells with a decrease in the expression of B-cell lymphoma-extra-large and (Bcl-xl) B-cell lymphoma 2 (Bcl-2), but an increase in the expression of B-cell lymphoma 2-associated X protein (Bax). Sch B promoted the cleavage of caspase 3 and poly-adenosine diphosphate-ribose polymerase (PARP) in both cell lines. Additionally, Sch B significantly induced autophagy of AML-12 and RAW 264.7 cells. Sch B inhibited the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway, as indicated by their altered phosphorylation, contributing to the proautophagic effect of Sch B. Taken together, our findings show that the inducing effects of Sch B on cell cycle arrest, apoptosis, and autophagy may contribute to its liver toxic effects, which might provide a clue for the investigation of the molecular toxic targets and underlying mechanisms for Sch B-induced hepatotoxicity in herbal consumers. More studies are warranted to fully delineate the underlying mechanisms, efficacy, and safety of Sch B for clinical use.

No MeSH data available.


Related in: MedlinePlus

Sch B modulates the expression of key pro- and anti-apoptotic molecules in mouse AML-12 and RAW 264.7 cells.Notes: Cells were treated with Sch B at 0.1, 1, 10, and 25 μM for 24 hours and then the protein samples were subject to Western blotting assay. (A) Representative blots of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in AML-12 and RAW 264.7 cells and Bar graphs showing the relative levels of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in mouse AML-12 (B) and RAW 264.7 cells (C). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; Bcl-2, B-cell lymphoma 2; Bcl-xl, B-cell lymphoma extra-large; Bax, Bcl-2-like protein4/Bcl-2-associated X protein; PARP, poly-adenosine diphosphate-ribose polymerase.
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f7-dddt-9-2001: Sch B modulates the expression of key pro- and anti-apoptotic molecules in mouse AML-12 and RAW 264.7 cells.Notes: Cells were treated with Sch B at 0.1, 1, 10, and 25 μM for 24 hours and then the protein samples were subject to Western blotting assay. (A) Representative blots of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in AML-12 and RAW 264.7 cells and Bar graphs showing the relative levels of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in mouse AML-12 (B) and RAW 264.7 cells (C). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; Bcl-2, B-cell lymphoma 2; Bcl-xl, B-cell lymphoma extra-large; Bax, Bcl-2-like protein4/Bcl-2-associated X protein; PARP, poly-adenosine diphosphate-ribose polymerase.

Mentions: Since we had observed the pro-apoptotic effect of Sch B in AML-12 and RAW 264.7 cells, we further investigated the underlying mechanism for Sch B-induced apoptosis in AML-12 and RAW 264.7 cells. We evaluated the expression levels of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in these two cell lines treated with Sch B at 0.1, 1, 10, and 25 μM for 24 hours. We first examined the effects of Sch B on the expression levels of the pro-apoptotic protein Bax, the anti-apoptotic proteins Bcl-2 and Bcl-xl, and the Bax/Bcl-2 ratio by using Western blotting analysis. Treatment of AML-12 and RAW 264.7 cells with Sch B at 0.1, 1, 10, and 25 μM increased the expression level of Bax but decreased the expression level of Bcl-2, resulting in a significant decrease in Bax/Bcl-2 ratio in a concentration-dependent manner (Figure 7A and B). Incubation of AML-12 cells with Sch B at both 10 and 25 μM significantly increased the level of Bax by 66.4% and 81.4%, respectively (P<0.05 by one-way ANOVA; Figure 7A and B). In contrast, the expression level of Bcl-2 was reduced by 17.4%, 26.4%, and 23.3% when AML-12 cells were treated with Sch B at 1, 10, and 25 μM, respectively (P>0.05 by one-way ANOVA; Figure 7A and B). The ratio of Bcl-2/Bax was significantly decreased 39.3%, 42.0%, and 58.5% when AML-12 cells were treated with Sch B at 1, 10, and 25 μM, respectively (P<0.05 or P<0.01 by one-way ANOVA; Figure 7A and B). Similarly, exposure of RAW 264.7 cells to Sch B at 1, 10, and 25 μM significantly increased the expression level of Bax by 98.8%, 116.0%, and 160.7%, respectively, compared with control cells (P<0.05 or P<0.01 by one-way ANOVA; Figure 7A and C), but decreased the level of Bcl-2 by 19.7%, 54.9%, 45.6%, and 59.6%, after cells were treated with Sch B at 0.1, 1, 10, and 25 μM, respectively. The Bcl-2/Bax ratio in RAW 264.7 cells was significantly reduced 53.7%, 78.5%, 75.2%, and 84.4% when treated with Sch B at 0.1, 1, 10, and 25 μM, respectively (P<0.01 or P<0.001 by one-way ANOVA; Figure 7A and C). In AML-12 cells, the expression level of Bcl-xl was remarkably decreased by 38.6% and 53.3% when treated with Sch B at 10 and 25 μM, respectively (P<0.05 by one-way ANOVA; Figure 7A and B). In RAW 264.7 cells, the expression level of Bcl-xl was slightly decreased, without statistical significance, when treated with Sch B for 24 hours (P>0.05 by one-way ANOVA; Figure 7A and C).


Schisandrin B inhibits cell growth and induces cellular apoptosis and autophagy in mouse hepatocytes and macrophages: implications for its hepatotoxicity.

Zhang Y, Zhou ZW, Jin H, Hu C, He ZX, Yu ZL, Ko KM, Yang T, Zhang X, Pan SY, Zhou SF - Drug Des Devel Ther (2015)

Sch B modulates the expression of key pro- and anti-apoptotic molecules in mouse AML-12 and RAW 264.7 cells.Notes: Cells were treated with Sch B at 0.1, 1, 10, and 25 μM for 24 hours and then the protein samples were subject to Western blotting assay. (A) Representative blots of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in AML-12 and RAW 264.7 cells and Bar graphs showing the relative levels of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in mouse AML-12 (B) and RAW 264.7 cells (C). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; Bcl-2, B-cell lymphoma 2; Bcl-xl, B-cell lymphoma extra-large; Bax, Bcl-2-like protein4/Bcl-2-associated X protein; PARP, poly-adenosine diphosphate-ribose polymerase.
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Related In: Results  -  Collection

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f7-dddt-9-2001: Sch B modulates the expression of key pro- and anti-apoptotic molecules in mouse AML-12 and RAW 264.7 cells.Notes: Cells were treated with Sch B at 0.1, 1, 10, and 25 μM for 24 hours and then the protein samples were subject to Western blotting assay. (A) Representative blots of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in AML-12 and RAW 264.7 cells and Bar graphs showing the relative levels of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in mouse AML-12 (B) and RAW 264.7 cells (C). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; Bcl-2, B-cell lymphoma 2; Bcl-xl, B-cell lymphoma extra-large; Bax, Bcl-2-like protein4/Bcl-2-associated X protein; PARP, poly-adenosine diphosphate-ribose polymerase.
Mentions: Since we had observed the pro-apoptotic effect of Sch B in AML-12 and RAW 264.7 cells, we further investigated the underlying mechanism for Sch B-induced apoptosis in AML-12 and RAW 264.7 cells. We evaluated the expression levels of Bcl-xl, Bcl-2, Bax, cytochrome c, cleaved PARP, and cleaved caspase 3 in these two cell lines treated with Sch B at 0.1, 1, 10, and 25 μM for 24 hours. We first examined the effects of Sch B on the expression levels of the pro-apoptotic protein Bax, the anti-apoptotic proteins Bcl-2 and Bcl-xl, and the Bax/Bcl-2 ratio by using Western blotting analysis. Treatment of AML-12 and RAW 264.7 cells with Sch B at 0.1, 1, 10, and 25 μM increased the expression level of Bax but decreased the expression level of Bcl-2, resulting in a significant decrease in Bax/Bcl-2 ratio in a concentration-dependent manner (Figure 7A and B). Incubation of AML-12 cells with Sch B at both 10 and 25 μM significantly increased the level of Bax by 66.4% and 81.4%, respectively (P<0.05 by one-way ANOVA; Figure 7A and B). In contrast, the expression level of Bcl-2 was reduced by 17.4%, 26.4%, and 23.3% when AML-12 cells were treated with Sch B at 1, 10, and 25 μM, respectively (P>0.05 by one-way ANOVA; Figure 7A and B). The ratio of Bcl-2/Bax was significantly decreased 39.3%, 42.0%, and 58.5% when AML-12 cells were treated with Sch B at 1, 10, and 25 μM, respectively (P<0.05 or P<0.01 by one-way ANOVA; Figure 7A and B). Similarly, exposure of RAW 264.7 cells to Sch B at 1, 10, and 25 μM significantly increased the expression level of Bax by 98.8%, 116.0%, and 160.7%, respectively, compared with control cells (P<0.05 or P<0.01 by one-way ANOVA; Figure 7A and C), but decreased the level of Bcl-2 by 19.7%, 54.9%, 45.6%, and 59.6%, after cells were treated with Sch B at 0.1, 1, 10, and 25 μM, respectively. The Bcl-2/Bax ratio in RAW 264.7 cells was significantly reduced 53.7%, 78.5%, 75.2%, and 84.4% when treated with Sch B at 0.1, 1, 10, and 25 μM, respectively (P<0.01 or P<0.001 by one-way ANOVA; Figure 7A and C). In AML-12 cells, the expression level of Bcl-xl was remarkably decreased by 38.6% and 53.3% when treated with Sch B at 10 and 25 μM, respectively (P<0.05 by one-way ANOVA; Figure 7A and B). In RAW 264.7 cells, the expression level of Bcl-xl was slightly decreased, without statistical significance, when treated with Sch B for 24 hours (P>0.05 by one-way ANOVA; Figure 7A and C).

Bottom Line: However, the potential hepatotoxicity of Sch B is a major safety concern, and the underlying mechanism for Sch B-induced liver toxic effects is not fully elucidated.The results have shown that Sch B exhibits potent grow inhibitory, proapoptotic, and proautophagic effects in AML-12 and RAW 264.7 cells.More studies are warranted to fully delineate the underlying mechanisms, efficacy, and safety of Sch B for clinical use.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, People's Republic of China ; Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.

ABSTRACT
A number of drugs and herbal compounds have been documented to cause hepatoxicity. Schisandrin B (Sch B) is an active dibenzocyclooctadiene isolated from Schisandrae fructus, with a wide array of pharmacological activities. However, the potential hepatotoxicity of Sch B is a major safety concern, and the underlying mechanism for Sch B-induced liver toxic effects is not fully elucidated. In the present study, we aimed to investigate the liver toxic effects and the molecular mechanisms of Sch B in mouse liver and macrophage cells. The results have shown that Sch B exhibits potent grow inhibitory, proapoptotic, and proautophagic effects in AML-12 and RAW 264.7 cells. Sch B markedly arrested cells in G1 phase in both cell lines, accompanied by the down-regulation of cyclin dependent kinase 2 (CDK2) and cyclin D1 and up-regulation of p27 Kip1 and checkpoint kinase 1. Furthermore, Sch B markedly increased the apoptosis of AML-12 and RAW 264.7 cells with a decrease in the expression of B-cell lymphoma-extra-large and (Bcl-xl) B-cell lymphoma 2 (Bcl-2), but an increase in the expression of B-cell lymphoma 2-associated X protein (Bax). Sch B promoted the cleavage of caspase 3 and poly-adenosine diphosphate-ribose polymerase (PARP) in both cell lines. Additionally, Sch B significantly induced autophagy of AML-12 and RAW 264.7 cells. Sch B inhibited the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway, as indicated by their altered phosphorylation, contributing to the proautophagic effect of Sch B. Taken together, our findings show that the inducing effects of Sch B on cell cycle arrest, apoptosis, and autophagy may contribute to its liver toxic effects, which might provide a clue for the investigation of the molecular toxic targets and underlying mechanisms for Sch B-induced hepatotoxicity in herbal consumers. More studies are warranted to fully delineate the underlying mechanisms, efficacy, and safety of Sch B for clinical use.

No MeSH data available.


Related in: MedlinePlus