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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 levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 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 subjected to Western blotting assay. Representative blots of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 (A). Bar graphs show the relative expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 in mouse AML-12 and RAW 264.7 cells (B). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05 and **P<0.01 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; CDK2, cyclin-dependent kinase 2; Chk1, checkpoint kinase 1; PCNA, proliferating cell nuclear antigen; E2F1, E2F transcription factor 1.
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f4-dddt-9-2001: Sch B modulates the expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 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 subjected to Western blotting assay. Representative blots of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 (A). Bar graphs show the relative expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 in mouse AML-12 and RAW 264.7 cells (B). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05 and **P<0.01 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; CDK2, cyclin-dependent kinase 2; Chk1, checkpoint kinase 1; PCNA, proliferating cell nuclear antigen; E2F1, E2F transcription factor 1.

Mentions: To further investigate the molecular mechanism for Sch B-induced cell cycle arrest in AML-12 and RAW 264.7 cells, the expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 in AML-12 and RAW 264.7 cells were examined by Western blotting assay. As shown in Figure 4A and B, Sch B treatment resulted in varying alterations in the expression of these key cell cycle regulators in both cell lines. Compared to the control cells, incubation of AML-12 cells with Sch B significantly altered the expression levels of cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1, whereas there were no significant changes in the expression levels of CDK2 and cyclin B1. There were 43.7% and 44.0% decreases in the expression level of cyclin D1 when AML-12 cells were treated with 10 and 25 μM Sch B, respectively (Figure 4A and B), and there was a 96.8% increase in the expression level of p27 Kip1 when AML-12 cells were treated with 25 μM Sch B (P<0.01 by one-way ANOVA; Figure 4A and B). Treatment of AML- 12 cells with Sch B at 1, 10, and 25 μM resulted in a 109.1%, 89.6%, and 69.2% increase in the expression level of Chk1 (P<0.05 or P<0.01 by one-way ANOVA). In RAW 264.7 cells, Sch B significantly altered the expression levels of p27 Kip1 and PCNA, whereas there was no significant alteration in the expression levels of CDK2, cyclin B1, cyclin D1, Chk1, and E2F1 (Figure 4A and B). There was a 251.9% increase in the expression level of p27 Kip1 when RAW 264.7 cells were treated with 25 μM Sch B (P<0.01 by one-way ANOVA). The expression level of cyclin D1 in RAW 264.7 cells was decreased by 19.3%, 5.6%, 24.6%, and 53.3% when cells were treated with Sch B at 0.1, 1, 10, and 25 μM, respectively, but there was no statistical significance compared to control cells (P>0.05 by one-way ANOVA; Figure 4A and B).


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 levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 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 subjected to Western blotting assay. Representative blots of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 (A). Bar graphs show the relative expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 in mouse AML-12 and RAW 264.7 cells (B). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05 and **P<0.01 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; CDK2, cyclin-dependent kinase 2; Chk1, checkpoint kinase 1; PCNA, proliferating cell nuclear antigen; E2F1, E2F transcription factor 1.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4403607&req=5

f4-dddt-9-2001: Sch B modulates the expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 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 subjected to Western blotting assay. Representative blots of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 (A). Bar graphs show the relative expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 in mouse AML-12 and RAW 264.7 cells (B). β-Actin was used as the internal control. Data are the mean ± SEM of three independent experiments. *P<0.05 and **P<0.01 by one-way ANOVA.Abbreviations: Sch B, schisandrin B; SEM, standard error of the mean; ANOVA, analysis of variance; CDK2, cyclin-dependent kinase 2; Chk1, checkpoint kinase 1; PCNA, proliferating cell nuclear antigen; E2F1, E2F transcription factor 1.
Mentions: To further investigate the molecular mechanism for Sch B-induced cell cycle arrest in AML-12 and RAW 264.7 cells, the expression levels of CDK2, cyclin B1, cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1 in AML-12 and RAW 264.7 cells were examined by Western blotting assay. As shown in Figure 4A and B, Sch B treatment resulted in varying alterations in the expression of these key cell cycle regulators in both cell lines. Compared to the control cells, incubation of AML-12 cells with Sch B significantly altered the expression levels of cyclin D1, p27 Kip1, Chk1, PCNA, and E2F1, whereas there were no significant changes in the expression levels of CDK2 and cyclin B1. There were 43.7% and 44.0% decreases in the expression level of cyclin D1 when AML-12 cells were treated with 10 and 25 μM Sch B, respectively (Figure 4A and B), and there was a 96.8% increase in the expression level of p27 Kip1 when AML-12 cells were treated with 25 μM Sch B (P<0.01 by one-way ANOVA; Figure 4A and B). Treatment of AML- 12 cells with Sch B at 1, 10, and 25 μM resulted in a 109.1%, 89.6%, and 69.2% increase in the expression level of Chk1 (P<0.05 or P<0.01 by one-way ANOVA). In RAW 264.7 cells, Sch B significantly altered the expression levels of p27 Kip1 and PCNA, whereas there was no significant alteration in the expression levels of CDK2, cyclin B1, cyclin D1, Chk1, and E2F1 (Figure 4A and B). There was a 251.9% increase in the expression level of p27 Kip1 when RAW 264.7 cells were treated with 25 μM Sch B (P<0.01 by one-way ANOVA). The expression level of cyclin D1 in RAW 264.7 cells was decreased by 19.3%, 5.6%, 24.6%, and 53.3% when cells were treated with Sch B at 0.1, 1, 10, and 25 μM, respectively, but there was no statistical significance compared to control cells (P>0.05 by one-way ANOVA; Figure 4A and B).

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