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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 cell apoptosis in a H22-bearing liver cancer mouse model. (A) At the end of the experiment, the tumor tissue samples from both the control and EAEHJ-treated groups were processed for immunohistochemical (IHC) staining using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Representative images were captured at a ×400 magnification. (B) Quantification of the IHC assay was measured as the percentage of positively-stained cells. Data are presented as the mean ± standard deviation (n=8). *P<0.01, vs. control. (C) Effects of EAEHJ on the apoptosis of HepG2 human hepatoma cells. HepG2 human hepatoma cells were collected and stained with Annexin V/propidium iodine followed by 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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f2-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on cell apoptosis in a H22-bearing liver cancer mouse model. (A) At the end of the experiment, the tumor tissue samples from both the control and EAEHJ-treated groups were processed for immunohistochemical (IHC) staining using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Representative images were captured at a ×400 magnification. (B) Quantification of the IHC assay was measured as the percentage of positively-stained cells. Data are presented as the mean ± standard deviation (n=8). *P<0.01, vs. control. (C) Effects of EAEHJ on the apoptosis of HepG2 human hepatoma cells. HepG2 human hepatoma cells were collected and stained with Annexin V/propidium iodine followed by 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: In order to determine whether the inhibitory effects of EAEHJ on tumor growth were due to apoptosis, the present study examined the EAEHJ pro-apoptotic activities in tumor-bearing mice via IHC TUNEL staining. The percentage of TUNEL-positive cells in the control and EAEHJ-treated mouse groups was 31.5±11 and 57.3±15%, respectively (P<0.01, Fig. 2A and B). These results indicate that EAEHJ may promote cell apoptosis in vivo.


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 cell apoptosis in a H22-bearing liver cancer mouse model. (A) At the end of the experiment, the tumor tissue samples from both the control and EAEHJ-treated groups were processed for immunohistochemical (IHC) staining using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Representative images were captured at a ×400 magnification. (B) Quantification of the IHC assay was measured as the percentage of positively-stained cells. Data are presented as the mean ± standard deviation (n=8). *P<0.01, vs. control. (C) Effects of EAEHJ on the apoptosis of HepG2 human hepatoma cells. HepG2 human hepatoma cells were collected and stained with Annexin V/propidium iodine followed by 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

f2-mmr-12-04-4851: Effects of Hypericum japonicum ethyl acetate extract (EAEHJ) on cell apoptosis in a H22-bearing liver cancer mouse model. (A) At the end of the experiment, the tumor tissue samples from both the control and EAEHJ-treated groups were processed for immunohistochemical (IHC) staining using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Representative images were captured at a ×400 magnification. (B) Quantification of the IHC assay was measured as the percentage of positively-stained cells. Data are presented as the mean ± standard deviation (n=8). *P<0.01, vs. control. (C) Effects of EAEHJ on the apoptosis of HepG2 human hepatoma cells. HepG2 human hepatoma cells were collected and stained with Annexin V/propidium iodine followed by 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: In order to determine whether the inhibitory effects of EAEHJ on tumor growth were due to apoptosis, the present study examined the EAEHJ pro-apoptotic activities in tumor-bearing mice via IHC TUNEL staining. The percentage of TUNEL-positive cells in the control and EAEHJ-treated mouse groups was 31.5±11 and 57.3±15%, respectively (P<0.01, Fig. 2A and B). These results indicate that EAEHJ may promote cell apoptosis in vivo.

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