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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 phosphorylation of PI3K, Akt, and mTOR, and the expression of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells.Notes: Bar graphs show the ratio of p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR, and the relative levels of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: p, phosphorylated; Sch B, schisandrin B; PI3K, phosphatidylinositol 3-kinase; Akt, protein kinase B; mTOR, mammalian target of rapamycin; AMPK, 5′-adenosine monophosphate-activated protein kinase; PTEN, phosphatase and tensin homolog; LC3-I, cytosolic microtubule-associated protein 1A/1B-light chain 3; LC3-II, membrane-bound LC3-phosphatidylethanolamine conjugate; SEM, standard error of the mean; ANOVA, analysis of variance.
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f11-dddt-9-2001: Sch B modulates the phosphorylation of PI3K, Akt, and mTOR, and the expression of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells.Notes: Bar graphs show the ratio of p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR, and the relative levels of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: p, phosphorylated; Sch B, schisandrin B; PI3K, phosphatidylinositol 3-kinase; Akt, protein kinase B; mTOR, mammalian target of rapamycin; AMPK, 5′-adenosine monophosphate-activated protein kinase; PTEN, phosphatase and tensin homolog; LC3-I, cytosolic microtubule-associated protein 1A/1B-light chain 3; LC3-II, membrane-bound LC3-phosphatidylethanolamine conjugate; SEM, standard error of the mean; ANOVA, analysis of variance.

Mentions: Next, we examined the effect of Sch B on the expression level of PI3K class III, beclin 1, LC3-I, and LC3-II. As shown in Figures 9 and 10, treatment of AML-12 cells with Sch B at 25 μM markedly increased the expression level of PI3K class III by 139.5%, compared to the control cells (P<0.001 by one-way ANOVA). There were 38.9%, 140.6%, 100.8%, and 161.5% increases in the expression level of PI3K class III in RAW 264.7 cells treated with 0.1, 1, 10, and 25 μM Sch B for 24 hours, respectively (P<0.01 by one-way ANOVA; Figures 9 and 11). After 24 hours treatment with Sch B at 25 μM, the expression level of beclin 1 in AML-12 cells was increased 62.3%, compared to the control cells (P<0.01 by one-way ANOVA; Figures 9 and 10). In RAW 264.7 cells, treatment with Sch B at 1, 10, and 25 μM resulted in a 71.5%, 120.0%, and 169.2% increase in the expression of beclin 1, respectively (P<0.05 or P<0.01 by one-way ANOVA; Figures 9 and 11). Furthermore, in the present study, our Western blotting analysis revealed two clear bands of LC3-I and LC3-II in RAW 264.7 cells after Sch B treatment. However, only one distinct band of LC3-I could be detected in AML-12 cells (Figure 9). Compared to control cells, treatment of AML-12 cells with Sch B at 10 μM and 25 μM for 24 hours significantly increased the level of LC3-I by 73.6% and 96.2%, respectively (P<0.05 or P<0.01 by one-way ANOVA; Figures 9 and 10). In RAW 264.7 cells, the expression level of LC3-I was markedly increased by 81.2%, 89.2%, and 166.1%, respectively, when treated with Sch B at 0.1, 10, and 25 μM for 24 hours (P<0.05 or P<0.01 by one-way ANOVA; Figures 9 and 11). In addition, the level of LC3-II was significantly increased 3.8-, 4.8-, and 3.6-fold when RAW 264.7 cells were treated with Sch B at 0.1, 10, and 25 μM, respectively. Treatment of RAW 264.7 cells with Sch B at 0.1, 1, 10, and 25 μM increased the ratio of LC3-II/LC3-I, although the difference was not significant (P>0.05 by one-way ANOVA; Figures 9 and 11).


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 phosphorylation of PI3K, Akt, and mTOR, and the expression of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells.Notes: Bar graphs show the ratio of p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR, and the relative levels of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: p, phosphorylated; Sch B, schisandrin B; PI3K, phosphatidylinositol 3-kinase; Akt, protein kinase B; mTOR, mammalian target of rapamycin; AMPK, 5′-adenosine monophosphate-activated protein kinase; PTEN, phosphatase and tensin homolog; LC3-I, cytosolic microtubule-associated protein 1A/1B-light chain 3; LC3-II, membrane-bound LC3-phosphatidylethanolamine conjugate; SEM, standard error of the mean; ANOVA, analysis of variance.
© Copyright Policy
Related In: Results  -  Collection

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

f11-dddt-9-2001: Sch B modulates the phosphorylation of PI3K, Akt, and mTOR, and the expression of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells.Notes: Bar graphs show the ratio of p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR, and the relative levels of AMPK, PTEN, PI3K class III, beclin 1, LC3-I, and LC3-II in mouse RAW 264.7 cells. Data are the mean ± SEM of three independent experiments. *P<0.05; **P<0.01; and ***P<0.001 by one-way ANOVA.Abbreviations: p, phosphorylated; Sch B, schisandrin B; PI3K, phosphatidylinositol 3-kinase; Akt, protein kinase B; mTOR, mammalian target of rapamycin; AMPK, 5′-adenosine monophosphate-activated protein kinase; PTEN, phosphatase and tensin homolog; LC3-I, cytosolic microtubule-associated protein 1A/1B-light chain 3; LC3-II, membrane-bound LC3-phosphatidylethanolamine conjugate; SEM, standard error of the mean; ANOVA, analysis of variance.
Mentions: Next, we examined the effect of Sch B on the expression level of PI3K class III, beclin 1, LC3-I, and LC3-II. As shown in Figures 9 and 10, treatment of AML-12 cells with Sch B at 25 μM markedly increased the expression level of PI3K class III by 139.5%, compared to the control cells (P<0.001 by one-way ANOVA). There were 38.9%, 140.6%, 100.8%, and 161.5% increases in the expression level of PI3K class III in RAW 264.7 cells treated with 0.1, 1, 10, and 25 μM Sch B for 24 hours, respectively (P<0.01 by one-way ANOVA; Figures 9 and 11). After 24 hours treatment with Sch B at 25 μM, the expression level of beclin 1 in AML-12 cells was increased 62.3%, compared to the control cells (P<0.01 by one-way ANOVA; Figures 9 and 10). In RAW 264.7 cells, treatment with Sch B at 1, 10, and 25 μM resulted in a 71.5%, 120.0%, and 169.2% increase in the expression of beclin 1, respectively (P<0.05 or P<0.01 by one-way ANOVA; Figures 9 and 11). Furthermore, in the present study, our Western blotting analysis revealed two clear bands of LC3-I and LC3-II in RAW 264.7 cells after Sch B treatment. However, only one distinct band of LC3-I could be detected in AML-12 cells (Figure 9). Compared to control cells, treatment of AML-12 cells with Sch B at 10 μM and 25 μM for 24 hours significantly increased the level of LC3-I by 73.6% and 96.2%, respectively (P<0.05 or P<0.01 by one-way ANOVA; Figures 9 and 10). In RAW 264.7 cells, the expression level of LC3-I was markedly increased by 81.2%, 89.2%, and 166.1%, respectively, when treated with Sch B at 0.1, 10, and 25 μM for 24 hours (P<0.05 or P<0.01 by one-way ANOVA; Figures 9 and 11). In addition, the level of LC3-II was significantly increased 3.8-, 4.8-, and 3.6-fold when RAW 264.7 cells were treated with Sch B at 0.1, 10, and 25 μM, respectively. Treatment of RAW 264.7 cells with Sch B at 0.1, 1, 10, and 25 μM increased the ratio of LC3-II/LC3-I, although the difference was not significant (P>0.05 by one-way ANOVA; Figures 9 and 11).

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