• 한국약용작물학회지
• /
• 제16권6호
• /
• pp.409-415
• /
• 2008

Moutan cortex, the root bark of Paeonia suffruticosa Andrews (Paeoniaceae), has pharmacological effects such as anti-inflammatory, antiallergic, analgesic and antioxidant activities. We investigated a methanol extract of Moutan cortex for neuroprotective effects on neurotoxicity induced by amyloid ${\beta}$ protein ($A{\beta}$) (25-35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to $10\;{\mu}M\;A{\beta}$ (25-35) for 24 h induced neuronal apoptotic death. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced neuronal cell death at 30 and $50\;{\mu}g/m{\ell}$, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), and generation of reactive oxygen species (ROS) which were measured by fluorescent dyes. Moutan cortex also inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}$ (25-35), which was measured by HPLC. These results suggest that Moutan cortex prevents $A{\beta}$ (25-35)-induced neuronal cell damage by interfering with the increase of $[Ca^{2+}]_i$, and then inhibiting glutamate release and ROS generation. Moutan cortex may have a therapeutic role in preventing the progression of Alzheimer's disease.

• 한국약용작물학회지
• /
• 제13권2호
• /
• pp.95-102
• /
• 2005

Caulis Bambusae in Taenia is widely used in Korea and China due to its various pharmacological activity. The present study aims to investigate the effect of the methanol extract of Caulis Bambusae in Taenia (CB) from Phyllostachys nigra Munro var. henonis Stapf (Gramineae) on amyloid ${\beta}$ protein (25-35) $(A{\beta}\;(25-35))$, a synthetic 25-35 amyloid peptide, -induced neurotoxicity using cultured rat cortical neurons. CB, over a concentration range of $10-50{\mu}g/{\mu}l$, inhibited the $A{\beta}\;(25-35)\;(10\;{\mu}M)$-induced neuronal cell death, as assessed by a 3-[4,5-dimethyIthiazole-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and the number of apoptotic nuclei, evidenced by Hoechst 33342 staining. CB $(50\;{\mu}g/{\mu}l)$ inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}$, (25-35) which was measured by HPLC. Pretreatment of CB $(50\;{\mu}g/{\mu}l)$ inhibited $10{\mu}M\;A{\beta}$ (25-35)-induced elevation of cytosolic calcium concentration $([Ca^{2+}]_c)$, which was measured by a fluorescent dye, fluo-4 AM, and generation of reactive oxygen species. These results suggest that CB prevents $A{\beta}$ (25-35)-induced neuronal ell damage in vitro.

• 한국약용작물학회지
• /
• 제15권6호
• /
• pp.444-450
• /
• 2007

Dried leaves from Cedrela sinensis A. Juss. (CS), have been observed to possess various pharmacological activity and contain various antioxidant constituents. The protective effect of ethanol extract of CS on hydrogen peroxide $(H_2O_2)-induced$ neurotoxicity was examined using primary cultured rat cortical neurons in the present study. Exposure of cultured neurons to 100 ${\mu}M\;H_2O_2$ caused a significant neuronal death as assessed by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. The addition of CS, over a concentration range of 10 to $50{\mu}g/m{\ell}$, concentration-dependently prevented the $H_2O_2-induced$ neuronal apoptotic death. CS $(50{\mu}g/m{\ell})$ significantly inhibited $H_2O_2-induced$ elevation of the cytosolic $Ca^{2+}$ concentration $([Ca^{2+}]_c)$, which was measured by a fluorescent dye, Fluo-4 AM. CS (30 and $50{\mu}g/m{\ell})$ inhibited glutamate release and generation of reactive oxygen species (ROS) induced by $100{\mu}M\;H_2O_2$. These results suggest that CS may mitigate the $H_2O_2-induced$ neurotoxiciy by interfering with the increase of $[Ca^{2+}]_c$, and then inhibiting glutamate release and generation of ROS in cultured neurons.

• Journal of Medicine and Life Science
• /
• 제15권2호
• /
• pp.67-71
• /
• 2018

Neuronal excitotoxicity induces mitochondrial dysfunction and the release of proapoptotic proteins. Excitotoxicity, the process by which the overactivation of excitatory neurotransmitter receptors leads to neuronal cell death. Neuronal death by excitotoxicity was related to neuronal degenerative disorders and hypoxia, results from excessive exposure to excitatory neurotransmitters, such as glutamate. Glutamate acts at NMDA receptors in cultured neurons to increase the intracellular free calcium concentration. Therefore endogenous glutamate may be a key factor to regulate neuronal cell death via activating $Ca^{2+}$ signaling. For this issue, we tested some conditions to alter intracellular $Ca^{2+}$ level in dissociated hippocampal neurons of rats. Cultured hippocampal neuron were treated by KCl (20 mM), $CaCl_2$ (3.8 mM) and glutamate ($5{\mu}M$) for 24 hrs. Interestingly, The Optical Density of hippocampal neurons was increased by high KCl application in MTT assay data. This enhanced response by high KCl was dependent on synaptic $Ca^{2+}$ influx but not on intracellular $Ca^{2+}$ level. However, the number of neurons seemed to be not changed in Hoechst 33342 staining data. These results suggest that enhancement of synaptic activity plays a key role to increase mitochondrial signaling in hippocampal neurons.

• Journal of Microbiology and Biotechnology
• /
• 제31권6호
• /
• pp.775-783
• /
• 2021

Sea cucumber, Holothuria scabra, is a well-known traditional Asian medicine that has been used for suppressing inflammation, promoting wound healing, and improving immunity. Moreover, previous studies demonstrated that the extract from H. scabra contains many bioactive compounds with potent inhibitory effect on tumor cell survival and progression. However, the effect of the methanolic extract from the body wall of H. scabra (BWMT) on human prostate cancer cells has not yet been investigated. In this study, we aimed to investigate the effects and underlying mechanism of BWMT on prostate cancer cell viability and metastasis. BWMT was obtained by maceration with methanol. The effect of BWMT on cell viability was assessed by MTT and colony formation assays. The intracellular ROS accumulation was evaluated using a DCFH-DA fluorescence probe. Hoechst 33342 staining and Annexin V-FITC/PI staining were used to examine the apoptotic-inducing effect of the extract. A transwell migration assay was performed to determine the anti-metastasis effect. BWMT significantly reduced cell viability and triggered cellular apoptosis by accumulating intracellular ROS resulting in the upregulation of JNK and p38 signaling pathways. In addition, BWMT also inhibited the invasion of PC3 cells by downregulating MMP-2/-9 expression via the ERK pathway. Consequently, our study provides BWMT from H. scabra as a putative therapeutic agent that could be applicable against prostate cancer progression.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권12호
• /
• pp.7179-7186
• /
• 2013

Background and Aims: Scutellaria is one of the most popular traditional Chinese herbal remedies against various human diseases, including cancer. In this study, we examined the active effects of Scutellaria extract and its main flavonoid constituents on the proportion of side population cells within human multiple myeloma cell line RPMI8226 in vitro and explored the potential molecular mechanisms involved. Materials and Methods: The contents of flavonoids in ethanolic extract of Scutellaria baicalensis Georgi were determined using high performance liquid chromatography. The antiproliferative effect of the ethanolic extract on RPMI-8226 was determined by CCK assay. Apoptosis was measured by annexin combining with propidium iodide in a flow cytometer. Cell cycle analysis was performed by propidium iodide staining in combination with flow cytometry analysis. Hoechst 33342 exclusion assay was used for the identification of side population within RPMI8226 cells. The expression of ABCG2 protein was assessed by Western blotting assay. Results: The content of major flavonoids constitutents of Scutellaria extract was baicalin (10.2%), wogonoside (2.50%), baicalein (2.29%), and wogonin (0.99%), respectively. The crude Scutellaria extract did not show significant anti-proliferative effect, apoptosis induction and cell cycle arrest in RPMI-8226 within the concentrations of $1-75{\mu}g/mL$. However, the ethanolic extract, baicalein, wogonin and baicalin reduced the side population cells in RPMI-8226, and data showed that baicalein and wogonin had stronger inhibitory effects. Correspondingly, they also exhibited significant effects on decreasing the expression level of ABCG2 protein in RPMI-8226 in vitro. Conclusions: Our results for the first time demonstrated a novel mechanism of action for Scutellaria extract and its main active flavonoids, namely targeting SP cells by modulating the expression of ABCG2 protein. This study provides an insight for new therapeutic strategies targeting cancer stem cells of multiple myeloma.

• 동의신경정신과학회지
• /
• 제20권2호
• /
• pp.19-29
• /
• 2009

Objectives : Antioxidant effects of Gagam-jangwonhwan(LMK01 and 02) water extract against $H_2O_2$-induced oxidative damage and cell death were investigated in rat pheochromocytoma line PC 12. Methods : The cells were treated with LMK01 and 02 water extract and $H_2O_2$, oxidative damage-inducing materials for 24 h. The cellular viability was assessed by WST-1 assay, oxidative damages of the cells by 8-OHdG quantitation, apoptosis by Hoechst 33342 staining assay and activity of antioxidant enzymes by catalase and glutathione peroxidase assay. Results : 1. LMK01 and LMK02 water extracts improved significantly cell viability in $H_2O_2$-treated groups than $H_2O_2$-alone treated cells 2, LMK02 suppressed significantly oxidative damage in $H_2O_2$-treated groups than $H_2O_2$-alone treated cells but LMK01 didn't. Meanwhile, difference of oxidative damages in conditions treated with LMK01 or LMK02 was not significant, 3. The $H_2O_2$ induced-apoptosis in PC 12 cell lines was inhibited effectively by LMK01 and LMK02, and especially the features of apoptosis were obviously reduced in LMK02-treated cells. 4. LMK01 and LMK02 increased significantly activities of both catalase and glutathione peroxidase than those of $H_2O_2$-alone treated group and moreover, LMK02 showed significantly higher activities than those of LMK01. Conclusions : As shown, LMK01 and LMK02 suppressed $H_2O_2$-induced oxidative damage and cell death in PC 12 cell effectively. And they increased activity of major antioxidant enzymes in PC 12 cell line. Therefore, this study suggests the possibility of clinical usage over oxidative stress-induced neurodegenerative disease such as Alzheimer's disease.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권9호
• /
• pp.4597-4606
• /
• 2012

Cholangiocarcinoma (CCA) is an uncommon adenocarcinoma which arises from the epithelial cells of the bile ducts. The aim of the study was to investigate the cytotoxicity, toxicity, and anticancer activity of a crude ethanolic extract of ginger (Zingiber officinale Roscoe) against CCA. Cytotoxic activity against a CCA cell line (CL-6) was assessed by calcein-AM and Hoechst 33342 assays and anti-oxidant activity was evaluated using the DPPH assay. Investigation of apoptotic activity was performed by DNA fragmentation assay and induction of genes that may be involved in the resistance of CCA to anticancer drugs (MDR1, MRP1, MRP2, and MRP3) was examined by real-time PCR. To investigate anti-CCA activity in vivo, a total of 80 OV and nitrosamine (OV/DMN)-induced CCA hamsters were fed with the ginger extract at doses of 1000, 3000, and 5000 mg/kg body weight daily or every alternate day for 30 days. Control groups consisting of 10 hamsters for each group were fed with 5-fluorouracil (positive control) or distilled water (untreated control). Median $IC_{50}$ (concentration that inhibits cell growth by 50%) values for cytotoxicity and anti-oxidant activities of the crude ethanolic extract of ginger were 10.95, 53.15, and $27.86{\mu}g/ml$, respectively. More than ten DNA fragments were visualized and up to 7-9 fold up-regulation of MDR1 and MRP3 genes was observed following exposure to the ethanolic extract of ginger. Acute and subacute toxicity tests indicated absence of any significant toxicity at the maximum dose of 5,000 mg/kg body weight given by intragastric gavage. The survival time and survival rate of the CCA-bearing hamsters were significantly prolonged compared to the control group (median of 54 vs 17 weeks). Results from these in vitro and in vivo studies thus indicate promising anticancer activity of the crude ethanolic extract of ginger against CCA with the absence of any significant toxicity. Moreover, MDR1 and MRP3 may be involved in conferring resistance of CCA to the ginger extract.

• 대한한방부인과학회지
• /
• 제24권3호
• /
• pp.14-27
• /
• 2011

Objectives: This study was designed to investigate the effects of Acanthopanacis Cortex Radicis extract(ACRE) on the apoptosis in HeLa cell and MCF-7 cell. Methods: After treatment with various concentration of ACRE, cell growth was evaluated in HeLa cell and MCF-7 cell. Hoechst 33342 staining was performed to estimate DNA fragment effect of ACRE on the apoptosis in HeLa cell and MCF-7 cell. Annexin V/PI apoptosis assay was used to estimate the effects of ACRE on the early apoptosis in HeLa cell and MCF-7 cell. RT-PCR was used to estimate the apoptosis gene expression effect of ACRE on Hela cell MCF-7 cell. Results: Under $0.1mg/m\ell$ of ACRE, cytotoxic effect was not found per NIH3T3 cell. The viability of HeLa cell and MCF-7 cells was significantly decreased ACRE ($100{\mu}g/m\ell$) in HeLa cell and MCF-7 cell, ACRE ($50{\mu}g/m\ell$) in HeLa cell 3 days after treatment, in MCF-7 cell 1&3 days after treatment (p<0.01). DNA fragmentation was observed 3 days after treatment of cl of ACRE on HeLa cell and MCF-7 cell. In Annexin V/PI apoptosis assay, after treatment of $100{\mu}g/m\ell$ of ACRE, the early apoptotic cell increased both in HeLa cell and MCF-7 cell. In RT-PCR analysis, after treatment of $100{\mu}g/m\ell$ of ACRE, bcl-2 were decreased and bax, caspase-3 were increased both in HeLa cell and MCF-7 cell. Conclusions: ACRE appears to have considerable activity on the apoptosis in HeLa cell and MCF-7 cell.

• 대한한방부인과학회지
• /
• 제24권3호
• /
• pp.1-13
• /
• 2011

Objectives: This study was designed to investigate the analysis of apoptosis by Scirpi Tuber in Hela cell and MCF-7 cell. Methods: For cytotoxic effect of Scirpi Tuber extract, Scirpi Tuber extract were cultured on NIH3T3 cell in vitro. After treatment with various concentration of Scirpi Tuber, cell growth was evaluated in Hela cell and MCF-7 cell. Hoechst 33342 staining was performed to estimate DNA fragment effect of Scirpi Tuber on the apoptosis in Hela cell and MCF-7 cell. Annexin V/PI apoptosis assay was used to estimate the effects of Scirpi Tuber on the early apoptosis in Hela cell MCF-7 cell. All the stained cells were analyzed by a FACS. RT-PCR was used to estimate the apoptosis gene expression effect of Scirpi Tuber extract on Hela cell and MCF-7 cell. Results: Cytotoxic effect of Scirpi Tuber extract was not found on per NIH3T3 cell. The viability of Hela cell was significantly decreased Scirpi Tuber (500, $1000{\mu}g/m\ell$) in Hela cell 1day, 3day and 5days after treatment (p<0.01). The viability of MCF-7 cell was significantly decresed Scirpi Tuber ($1000{\mu}g/m\ell$) in MCF-7 cell (p<0.01), Scirpi Tuber ($500{\mu}g/m\ell$) in MCF-7 cell only 3days after treatment (p<0.01). In RT-PCR analysis, after treatment of $100{\mu}g/m\ell$ of ACR extract, BCL-2 were decreased and BAX, caspase-3 were increased both in Hela cell and MCF-7 cell. DNA fragmentation was observed the Scirpi Tuber on Hela cell and MCF-7 cell. As time goes on DNA fragmentation incresed. In Annexin V/PI apoptosis assay, after treatment of $1mg/m\ell$ of Scirpi Tuber, the early apoptotic cell increased both in Hela cell and MCF-7 cell. As time goes on apoptotic cell increased. Conclusion: Scirpi Tuber appears to have considerable activity on the apoptosis in Hela cell and MCF-7 cell.