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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 cellular morphological changes and cell viability of HepG2 human hepatoma cells. The HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, and cellular morphological changes were subsequently observed using phase-contrast microscopy. The images were captured at a ×100 magnification for each experiment performed in triplicate. Cell viability was assessed using an MTT assay. The MTT assay indicated that EAEHJ treatment significantly reduced HepG2 human hepatoma cell viability in a dose-dependent manner. The data are presented as the mean ± standard deviation from three independent experiments. *P<0.01, vs. the control cells (100%, treated with 0.5% dimethyl sulphoxide vehicle).
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f1-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on cellular morphological changes and cell viability of HepG2 human hepatoma cells. The HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, and cellular morphological changes were subsequently observed using phase-contrast microscopy. The images were captured at a ×100 magnification for each experiment performed in triplicate. Cell viability was assessed using an MTT assay. The MTT assay indicated that EAEHJ treatment significantly reduced HepG2 human hepatoma cell viability in a dose-dependent manner. The data are presented as the mean ± standard deviation from three independent experiments. *P<0.01, vs. the control cells (100%, treated with 0.5% dimethyl sulphoxide vehicle).

Mentions: In order to examine the cytotoxicity and biological effects of EAEHJ, HepG2 human hepatoma cells were treated with various doses of EAEHJ (0, 0.5, 0.75, 1.5 and 3.0 mg/ml) for 24 h. Any cellular morphological changes were subsequently evaluated by phase-contrast microscopy. As shown in Fig. 1, the control cells exhibited a typical polygonal intact shape, whereas the cells treated with various concentrations of EAEHJ for 24 h became rounded, shrunken, exhibited membrane blebbing, or became non-confluent. Furthermore, the effects of the various concentrations of EAEHJ on HepG2 human hepatoma cell viability were examined using an MTT assay. The results were concordant with the microscopy results indicating that EAEHJ inhibited cell growth in a dose-dependent manner. At a concentration of 0.5 mg/ml, a significant loss of cell viability was observed. The cell growth inhibition rate was ~50% at 0.75 mg/ml EAEHJ (P<0.01). Upon incubation with 3.0 mg/ml EAEHJ, the cells were only 40% viable, (P<0.01, Fig. 1). These results indicate that EAEHJ is able to inhibit the growth of HepG2 human hepatoma cells in vitro.


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 cellular morphological changes and cell viability of HepG2 human hepatoma cells. The HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, and cellular morphological changes were subsequently observed using phase-contrast microscopy. The images were captured at a ×100 magnification for each experiment performed in triplicate. Cell viability was assessed using an MTT assay. The MTT assay indicated that EAEHJ treatment significantly reduced HepG2 human hepatoma cell viability in a dose-dependent manner. The data are presented as the mean ± standard deviation from three independent experiments. *P<0.01, vs. the control cells (100%, treated with 0.5% dimethyl sulphoxide vehicle).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on cellular morphological changes and cell viability of HepG2 human hepatoma cells. The HepG2 human hepatoma cells were treated with various concentrations of EAEHJ for 24 h, and cellular morphological changes were subsequently observed using phase-contrast microscopy. The images were captured at a ×100 magnification for each experiment performed in triplicate. Cell viability was assessed using an MTT assay. The MTT assay indicated that EAEHJ treatment significantly reduced HepG2 human hepatoma cell viability in a dose-dependent manner. The data are presented as the mean ± standard deviation from three independent experiments. *P<0.01, vs. the control cells (100%, treated with 0.5% dimethyl sulphoxide vehicle).
Mentions: In order to examine the cytotoxicity and biological effects of EAEHJ, HepG2 human hepatoma cells were treated with various doses of EAEHJ (0, 0.5, 0.75, 1.5 and 3.0 mg/ml) for 24 h. Any cellular morphological changes were subsequently evaluated by phase-contrast microscopy. As shown in Fig. 1, the control cells exhibited a typical polygonal intact shape, whereas the cells treated with various concentrations of EAEHJ for 24 h became rounded, shrunken, exhibited membrane blebbing, or became non-confluent. Furthermore, the effects of the various concentrations of EAEHJ on HepG2 human hepatoma cell viability were examined using an MTT assay. The results were concordant with the microscopy results indicating that EAEHJ inhibited cell growth in a dose-dependent manner. At a concentration of 0.5 mg/ml, a significant loss of cell viability was observed. The cell growth inhibition rate was ~50% at 0.75 mg/ml EAEHJ (P<0.01). Upon incubation with 3.0 mg/ml EAEHJ, the cells were only 40% viable, (P<0.01, Fig. 1). These results indicate that EAEHJ is able to inhibit the growth of HepG2 human hepatoma cells in vitro.

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