Limits...
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.

Show MeSH

Related in: MedlinePlus

Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the activity of caspases-3 and 9 in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h. (A) Caspase-9 and (B) caspase-3 activity levels were determined using a colorimetric assay. The data were normalized to the caspase activities within the control cells (treated with 0.5% dimethyl sulfoxide vehicle) and represented as the “fold of control”. 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
getmorefigures.php?uid=PMC4581784&req=5

f5-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the activity of caspases-3 and 9 in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h. (A) Caspase-9 and (B) caspase-3 activity levels were determined using a colorimetric assay. The data were normalized to the caspase activities within the control cells (treated with 0.5% dimethyl sulfoxide vehicle) and represented as the “fold of control”. The data are presented as the mean ± standard deviation from three independent experiments. *P< 0.01, vs. the control cells.

Mentions: Caspases form part of the cysteine protease family, and are important proteins in the modulation of the apoptotic response. Caspase-3 is a key regulator of apoptosis, and is activated by an initiator caspase, such as caspase-9, during mitochondria-mediated apoptosis (17). In order to identify the downstream effectors of the apoptotic signaling pathway, the activation of caspase-3 and 9 was examined using a colorimetric assay with the following caspase-specific chromophores: DEVD-pNA, a specific substrate of caspase-3, and LEHD-pNA, a specific substrate of caspase-9. As shown in Figs. 5A and B, treatment with EAEHJ significantly and dose-dependently induced activation of both caspase-3 and 9 in the HepG2 human hepatoma cells (P<0.01), as compared with the untreated control cells. These results suggest that EAEHJ is able to promote HepG2 human hepatoma cell apoptosis via the mitochondria-dependent pathway.


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 activity of caspases-3 and 9 in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h. (A) Caspase-9 and (B) caspase-3 activity levels were determined using a colorimetric assay. The data were normalized to the caspase activities within the control cells (treated with 0.5% dimethyl sulfoxide vehicle) and represented as the “fold of control”. 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

f5-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on the activity of caspases-3 and 9 in HepG2 human hepatoma cells. HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h. (A) Caspase-9 and (B) caspase-3 activity levels were determined using a colorimetric assay. The data were normalized to the caspase activities within the control cells (treated with 0.5% dimethyl sulfoxide vehicle) and represented as the “fold of control”. The data are presented as the mean ± standard deviation from three independent experiments. *P< 0.01, vs. the control cells.
Mentions: Caspases form part of the cysteine protease family, and are important proteins in the modulation of the apoptotic response. Caspase-3 is a key regulator of apoptosis, and is activated by an initiator caspase, such as caspase-9, during mitochondria-mediated apoptosis (17). In order to identify the downstream effectors of the apoptotic signaling pathway, the activation of caspase-3 and 9 was examined using a colorimetric assay with the following caspase-specific chromophores: DEVD-pNA, a specific substrate of caspase-3, and LEHD-pNA, a specific substrate of caspase-9. As shown in Figs. 5A and B, treatment with EAEHJ significantly and dose-dependently induced activation of both caspase-3 and 9 in the HepG2 human hepatoma cells (P<0.01), as compared with the untreated control cells. These results suggest that EAEHJ is able to promote HepG2 human hepatoma cell apoptosis via the mitochondria-dependent pathway.

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