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Ethyl acetate extract of Hypericum japonicum induces apoptosis via the mitochondria-dependent pathway in vivo and in vitro.

Zhuang Q, Li J, Chen Y, Lin J, Lai F, Chen X, Lin X, Peng J - Mol Med Rep (2015)

Bottom Line: Treatment with EAEHJ significantly reduced tumor weight, but had no effect on murine body weight.The results of the present study also showed that EAEHJ induced H22 cell apoptosis in vivo.Treatment with EAEHJ also increased the ratio of pro‑apoptotic B‑cell lymphoma 2 (Bcl‑2)‑associated X protein (Bax) to anti‑apoptotic Bcl‑2, and activated the caspase‑9 signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.

ABSTRACT
The widely-used Chinese medicinal herb Hypericum japonicum, also known as Hypericum japonicum Thunb or Tianjihuang, displays potent anti‑carcinogenic effects against liver cancer. However, the molecular mechanism underlying the therapeutic effects of Hypericum japonicum remains to be elucidated. The present study investigated the in vivo efficacy of ethyl acetate extract of Hypericum japonicum (EAEHJ) against tumor growth in an H22 cell‑bearing liver cancer mouse model. Treatment with EAEHJ significantly reduced tumor weight, but had no effect on murine body weight. The results of the present study also showed that EAEHJ induced H22 cell apoptosis in vivo. In addition, the anti‑carcinogenic effects of EAEHJ were investigated in vitro. The results of the present study demonstrate that both phospholipid asymmetry in the plasma membrane and mitochondrial membrane potential were deregulated in HepG2 human hepatoma cells, following treatment with EAEHJ. Treatment with EAEHJ also increased the ratio of pro‑apoptotic B‑cell lymphoma 2 (Bcl‑2)‑associated X protein (Bax) to anti‑apoptotic Bcl‑2, and activated the caspase‑9 signaling pathway. These results suggest that EAEHJ is able to trigger the apoptosis of liver cancer cells via the mitochondria-dependent pathway.

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Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the loss of mitochondrial membrane potential in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, prior to being stained with JC-1. The mean JC-1 fluorescence intensity was detected using fluorescence-activated cell sorting analysis. The data are presented as the mean ± standard deviation from three independent experiments, *P<0.01, vs. the control cells.
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f3-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the loss of mitochondrial membrane potential in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, prior to being stained with JC-1. The mean JC-1 fluorescence intensity was detected using fluorescence-activated cell sorting analysis. The data are presented as the mean ± standard deviation from three independent experiments, *P<0.01, vs. the control cells.

Mentions: Early-stage cellular apoptosis is accompanied by disruption of the mitochondrial membrane, which leads to a rapid collapse of the electrochemical gradient. In the present study, the effects of EAEHJ on ΔΨm were investigated using JC-1, a mitochondria-specific dye. JC-1 is a lipophilic, cationic dye that selectively enters mitochondria. In healthy cells with high ΔΨm, JC-1 forms J-aggregates that exhibit intense red fluorescence (detected in PMT3, 590 nm), whereas under apoptotic conditions, the ΔΨm collapses, and JC-1 does not accumulate within the mitochondria, but remains instead in the cytoplasm in its monomeric form, emitting green fluorescence (detected in PMT2, 525 nm). Flow cytometric analysis demonstrated the loss of ΔΨm in cells treated with EAEHJ for 24 h in a dose-dependent manner. As shown in Fig. 3, following treatment with 0, 0.5, 0.75, 1.5, and 3.0 mg/ml of EAEHJ, the ratio of PMT3 to PMT2 was 95.8%, 68.2%, 56%, 52.2%, and 23.3%, respectively (P<0.01). This dose-dependent change in ΔΨm indicates that EAEHJ promotes the loss of ΔΨm in HepG2 human hepatoma cells.


Ethyl acetate extract of Hypericum japonicum induces apoptosis via the mitochondria-dependent pathway in vivo and in vitro.

Zhuang Q, Li J, Chen Y, Lin J, Lai F, Chen X, Lin X, Peng J - Mol Med Rep (2015)

Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the loss of mitochondrial membrane potential in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, prior to being stained with JC-1. The mean JC-1 fluorescence intensity was detected using fluorescence-activated cell sorting analysis. The data are presented as the mean ± standard deviation from three independent experiments, *P<0.01, vs. the control cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581784&req=5

f3-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the loss of mitochondrial membrane potential in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, prior to being stained with JC-1. The mean JC-1 fluorescence intensity was detected using fluorescence-activated cell sorting analysis. The data are presented as the mean ± standard deviation from three independent experiments, *P<0.01, vs. the control cells.
Mentions: Early-stage cellular apoptosis is accompanied by disruption of the mitochondrial membrane, which leads to a rapid collapse of the electrochemical gradient. In the present study, the effects of EAEHJ on ΔΨm were investigated using JC-1, a mitochondria-specific dye. JC-1 is a lipophilic, cationic dye that selectively enters mitochondria. In healthy cells with high ΔΨm, JC-1 forms J-aggregates that exhibit intense red fluorescence (detected in PMT3, 590 nm), whereas under apoptotic conditions, the ΔΨm collapses, and JC-1 does not accumulate within the mitochondria, but remains instead in the cytoplasm in its monomeric form, emitting green fluorescence (detected in PMT2, 525 nm). Flow cytometric analysis demonstrated the loss of ΔΨm in cells treated with EAEHJ for 24 h in a dose-dependent manner. As shown in Fig. 3, following treatment with 0, 0.5, 0.75, 1.5, and 3.0 mg/ml of EAEHJ, the ratio of PMT3 to PMT2 was 95.8%, 68.2%, 56%, 52.2%, and 23.3%, respectively (P<0.01). This dose-dependent change in ΔΨm indicates that EAEHJ promotes the loss of ΔΨm in HepG2 human hepatoma cells.

Bottom Line: Treatment with EAEHJ significantly reduced tumor weight, but had no effect on murine body weight.The results of the present study also showed that EAEHJ induced H22 cell apoptosis in vivo.Treatment with EAEHJ also increased the ratio of pro‑apoptotic B‑cell lymphoma 2 (Bcl‑2)‑associated X protein (Bax) to anti‑apoptotic Bcl‑2, and activated the caspase‑9 signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.

ABSTRACT
The widely-used Chinese medicinal herb Hypericum japonicum, also known as Hypericum japonicum Thunb or Tianjihuang, displays potent anti‑carcinogenic effects against liver cancer. However, the molecular mechanism underlying the therapeutic effects of Hypericum japonicum remains to be elucidated. The present study investigated the in vivo efficacy of ethyl acetate extract of Hypericum japonicum (EAEHJ) against tumor growth in an H22 cell‑bearing liver cancer mouse model. Treatment with EAEHJ significantly reduced tumor weight, but had no effect on murine body weight. The results of the present study also showed that EAEHJ induced H22 cell apoptosis in vivo. In addition, the anti‑carcinogenic effects of EAEHJ were investigated in vitro. The results of the present study demonstrate that both phospholipid asymmetry in the plasma membrane and mitochondrial membrane potential were deregulated in HepG2 human hepatoma cells, following treatment with EAEHJ. Treatment with EAEHJ also increased the ratio of pro‑apoptotic B‑cell lymphoma 2 (Bcl‑2)‑associated X protein (Bax) to anti‑apoptotic Bcl‑2, and activated the caspase‑9 signaling pathway. These results suggest that EAEHJ is able to trigger the apoptosis of liver cancer cells via the mitochondria-dependent pathway.

Show MeSH
Related in: MedlinePlus