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Chemical Composition and Antiproliferative Activity of Supercritical Extract of Immature Citrus Peel in human cervical carcinoma HeLa cells (미성숙 감귤 과피 초임계 추출물의 성분 분석과 자궁암세포 성장억제효능)

  • Moon, Jeong Yong;Song, YeonWoo;Hyun, Ho Bong;KimCho, Somi
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.12
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    • pp.8836-8843
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    • 2015
  • This study was performed to investigate the antiproliferative activities of supercritical extracts from phalsak(Citrus hassaku Hort ex Tanaka) and yeagam(Citrus iyo Hort. ex Tanaka) against human cervical carcinoma HeLa cells and the chemical compositions of the extracts. The anticancer properties of supercritical extracts were demonstrated using the MTT assay and Hoechst 33342 staining and the compositional analyses were conducted by using gas chromatography-mass spectrometry(GC-MS). The peel extracts of both species exhibited similar antiproliferative effect. The antiproliferative activity of the flesh extracts was not detected up to $400{\mu}g/mL$, whereas peel extracts of phalsak and yeagam reduced cell viability with 87.16% and 92.95% at $400{\mu}g/mL$, respectively. There was a dramatic increase of the apoptotic body formation in the cell treated with peel extracts while no apoptotic body formation detected in the cell treated with flesh extracts at 100, $200{\mu}g/mL$. By GC-MS analysis, 27 and 31 kinds of compounds identified in flesh and peel of phalsak, while 27 and 29 kinds of compounds were identified in flesh and peel of yeagam, respectively. 1,1,4,4-Tetramethyl-2-tetralone(20.86%), alloimperatorin(8.15%), limonene(11.23%), and auraptene(7.29%) were major in peel of phalsak, whereas limonene(22.19%), linalool(11.23%), and ${\gamma}$-sitosterol(9.12%) were major in peel of yeagam.

Hepatoprotective and Anticancer Activities of Allomyrina dichotoma Larvae (장수풍뎅이 유충의 간보호 효능 및 항암활성)

  • Lee, Ji-Eun;Jo, Da-Eun;Lee, An-Jung;Park, Hye-Kyung;Youn, Kumju;Yun, Eun-Young;Hwang, Jae-Sam;Jun, Mira;Kang, Byoung Heon
    • Journal of Life Science
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    • v.25 no.3
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    • pp.307-316
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    • 2015
  • Beetle larvae have been used as a traditional medicine to treat various human liver diseases. To prove the liver protective function of Allomyrina dichotoma larvae (ADL), we induced liver damage by the intraperitoneal injection of a hepatotoxic reagent, diethylnitrosamine (DEN), to C3H/HeN male mice and orally administered freeze-dried ADL powder. ADL powder lessened DEN-induced hepatotoxicity considering the reduced signs of acute and chronic hepatotoxicities, such as the ALP level in the blood serum, TUNEL-positive hepatocytes, ductural reactions, steatotic hepatocytes, and collagen deposition of the Masson’s trichrome staining. In addition to hepatoprotection, the anti-cancer activity of ADL has been examined. The ADL powder was extracted with ethanol and then fractionated with hexane, ethyl acetate, and water by a solvent partition technique. The ethyl acetate fraction showed cytotoxicity to various cancer cells through induction of apoptosis and necrosis, as well as the perturbed metabolism of the cancer cell to trigger autophagy. Collectively, ADL contains bioactive substances that can protect hepatocytes from toxic chemicals and trigger cell death in cancer cells. Thus, further purification and analyses of ADL fractions could lead to the identification of novel bioactive compounds.

Protective Effect of Aqueous Extracts of Styela Clava Tunic Against Apoptosis of HepG2 Cells Induced by Hydrogen Peroxide (미더덕껍질의 유수추출물이 과산화수소에 의해 유발된 HepG2간암세포의 세포사멸에 미치는 보호 효과)

  • Koh, Eun Kyoung;Lee, Young Ju;Kim, Ji Eun;Kwak, Moon Hwa;Go, Jun;Son, Hong Joo;Lee, Hee Seob;Jung, Young Jin;Hwang, Dae Youn
    • Journal of Life Science
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    • v.24 no.6
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    • pp.595-602
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    • 2014
  • Styela Clava tunic (SCT) has found some applications in many areas of medical treatment including as an anti-inflammatory compound, a wound healing film, in guided bone regeneration, and as a food additive. The protective effect of SCT aqueous extract (AE-SCT) on cell death induced by $H_2O_2$ treatment was investigated by measuring the changes in cell viability in HepG2 cells after AE-SCT treatment. High concentrations of antioxidant compounds including flavonoids (3.3 mg/g) and phenolics (32.3 mg/g) were detected in AE-SCT but no significant cytotoxicity was observed in HepG2 cells treated with AE-SCT. The viability of HepG2 cells was also not changed by treatment with different concentrations of AE-SCT after $H_2O_2$ treatment. However, cell viability was significantly increased in cells treated with three different concentrations of AE-SCT before $H_2O_2$ treatment. The greatest increase in cell viability was observed in the group treated with $50{\mu}g/ml$ AE-SCT, when compared with vehicle-treated group. FACS and DAPI staining analysis indicated that the decrease in number of dead cells was dependent on the concentration of AE-SCT. Alterations in the Bax/Bcl-2 ratio after $H_2O_2$ treatment were significantly restored by treatment with different concentrations of AE-SCT. These results indicate that AE-SCT, which contains high levels of antioxidants, may protect cells against death induced by $H_2O_2$ treatment.

Nutritional Composition and Cytoprotective Effect of Moringa oleifera Lam. (Drumstick-tree (Moringa oleifera Lam.)의 주요 영양성분 및 추출물의 신경세포 보호 효과)

  • Jin, Su Il;Kim, Hyeon Ju;Jeong, Ji Hee;Jin, Dong Eun;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.609-615
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    • 2014
  • The cytoprotective effect of Moringa oleifera Lam. (drumstick tree) on neuronal cells was investigated to confirm the physiological benefits associated with this natural food resource. First, the drumstick tree extract was chemically analyzed to determine inherent nutritional constituents. Calcium and potassium were identified as the major mineral constituents, and palmitic acid (C16:0, 16.33%) and gadoleic acid (C20:01, 66.34%) were detected as the major fatty acids. Moreover, drumstick tree extract contained 94.78 mg/100 g vitamin E and 112.61 mg/100 g niacin. PC12 cells were used to study the cytoprotective effects of drumstick tree extract. Intracellular accumulation of reactive oxygen species was significantly reduced when $H_2O_2$ treated-neuronal cells were cultured in a medium containing the methanolic extract of drumstick tree, compared to cells treated with only $H_2O_2$. Cell viability assay using MTT showed that the extract protected cells against $H_2O_2$-induced neurotoxicity and inhibited LDH leakage from the cell membrane. Caspase assay showed that the extract exerted cytoprotective effect against apoptosis. Consequently, these data suggest that drumstick tree is a useful natural resource with positive effects on human health.

hCG-induced Endoplasmic Reticulum Stress Leads to Activation of the IRE1/XBP1 Pathway in Mouse Leydig Tumor Cells (mLTC-1) (mLTC-1 세포에 hCG 처리에 의해 유도된 소포체 스트레스가 IRE1/XBP1 경로의 활성화 유발)

  • Park, Sun-Ji;Kim, Tae-Shin;Lee, Dong-Seok
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1039-1045
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    • 2014
  • This study analyzed whether human chorionic gonadotropin (hCG) induces ER stress via the IRE/XBP1 pathway in mouse Leydig tumor (mLTC-1) cells. In a previous study, we demonstrated that the unfolding protein response (UPR) plays an important role in the expression of steroidogenic enzymes by modulating the ATF6 pathway, as well as ER stress-mediated apoptosis in hCG-stimulated Leydig cells. Although UPR signaling has been reported to regulate the IRE1/XBP1 pathway, it is not known whether hCG-induced ER stress in Leydig cells can activate the pathway. To investigate the activation of the IRE1/XBP1 pathway in mLTC-1 cells after hCG treatment, we performed a Western blot analysis to detect the phospho-IRE1 protein and an RT-PCR analysis to validate splicing of XBP1 mRNA. We used ER stress-activated indicator (ERAI) constructs for monitoring the activity of IRE1 and then analyzed by fluorescence microscopy and flow cytometry. The expression levels of the phospho-IRE1 protein markedly increased in response to the hCG treatment. In the mLTC-1 cells transfected with an F-XBP1-venus/F-$XBP1{\Delta}DBD$-venus construct, the hCG treatment led to the appearance of green fluorescent cells and detectable fluorescence in the nucleus and cytosol, respectively. In addition, splicing of XBP1 mRNA significantly increased after the hCG treatment. Taken together, these results indicate that hCG-induced ER stress leads to activation of the IRE1/XBP pathway in Leydig cells.

Cytotoxic Effects of Tenebrio molitor Larval Extracts against Hepatocellular Carcinoma (갈색거저리 유충 추출물의 간암세포에 대한 세포독성 효능)

  • Lee, Ji-Eun;Lee, An-Jung;Jo, Da-Eun;Cho, Ju Hyeong;Youn, Kumju;Yun, Eun-Young;Hwang, Jae-Sam;Jun, Mira;Kang, Byoung Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.2
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    • pp.200-207
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    • 2015
  • Various natural products or their derivatives, mostly originating from plants, fungi, and bacteria, have been exploited as therapeutic drugs to treat various human diseases. In addition to previously explored organisms, research on natural compounds has now expanded into unexamined living organisms in order to identify novel bioactive substances. Here, we determined whether or not the larval form of the mealworm beetle Tenebrio molitor, a species of darkling beetle, contains cytotoxic substances that exclusively affect cancer cell viability. Ethanol extract and its solvent partitioned fractions, hexane and ethyl acetate fractions, showed anticancer effects against various human cancer cells derived from the prostate (PC3 and 22Rv1), cervix (HeLa), liver (PLC/PRF5, HepG2, Hep3B, and SK-HEP-1), colon (HCT116), lung (NCI-H460), breast (MDA-MB231), and ovary (SKOV3). Cell death induced by the fractions was a mix of apoptosis, necrosis, and autophagy. The hexane fraction was administered intraperitoneally to nude mice bearing a hepatocellular carcinoma SK-HEP-1 and showed inhibition of tumor growth in vivo. Therefore, we concluded that worm extracts contain cytotoxic substances, which can be enriched by proper fractionation protocols, and further separation and purification could lead to the identification of novel molecules to treat human cancers.

Effects of Various Chitosan Oligomer Molecular Weight Levels on the Disorders of Lipid Metabolism and Immune-related Factors in Rats Treated 2,3,7,8-Tetrachlorodibenzo-p-dioxin (다이옥신계 TCDD(2,3,7,8-Tetrachlorodibenzo-p-dioxin)에 노출된 흰쥐의 지질대사 및 면역관련 인자에 대하여 키토산 올리고머의 분자량별 섭취효과)

  • Lee, Joon-Ho;Hwang, Seok-Youn;Lim, Beong-Ou;Lee, Yeon-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.4
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    • pp.471-479
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    • 2012
  • This study was conducted to investigate the effects of various levels of chitosan oligomer (CO) molecular weight on the disorders of lipid metabolism and immune-related factors induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), that is a endocrine disrupter, using adult male rats (SD) for 3 weeks. These 40 animals were divided into five groups. Three kinds of CO were used by molecular weight (MW) (less than 1000, 1000~3000, and 5000~10000) and added 4% to basal diets respectively. TCDD (40 ${\mu}g$/kg B.W) was intraperitoneally injected into rats at the beginning of the experiment. The relative weights of the livers were increased in all rats treated with TCDD, and the brain and testis weights were increased in all CO diet groups, compared to the control and TCDD groups. The levels of white blood cells (WBC) and red blood cells (RBC), hemoglobin, hematocrits (HCT), and platelets were significantly lowered by treating TCDD. By the way, RBC and HCT tended to recover by CO diets. The elevation of serum total and HDL cholesterol levels induced by TCDD treatment was significantly reduced by CO (5000~10000 MW) diets. The apparent increasing of the total lipid, cholesterol, and triglyceride levels of rat livers induced by TCDD was tended to be suppressed in those fed CO diets. Especially, diets with less than 1000 MW significantly diminished liver triglycerides. The levels of serum immunoglobulin (Ig) A, IgG1 and IgM were significantly high in rats fed CO (5000~10000 MW) diets. The decreasing levels of IgE by treatment with TCDD tended to recover all the CO diet groups to the level of control group. In histochemical observation, the fat droplets and apoptosis of liver due to TCDD treatment were markedly alleviated in all CO diet groups. These results indicated that CO, though not regular according to molecular weight, can exert improving effects on lipid accumulation, hepatocytic disorders, abnormal blood cells, and some immunoglobulins induced by TCDD.

Anti-Inflammatory Effects of Poly-${\gamma}$-Glutamic Acid on DNCB-Induced Allergic Contact Dermatitis in Dogs (개에서 DNCB에 의해 유발된 알레르기성 접촉피부염에 대한 폴리감마글루탐산의 항염증 효과)

  • Kim, Hyun-Gon;Kim, Kil-Soo;Oh, Tae-Ho
    • Journal of Veterinary Clinics
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    • v.29 no.4
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    • pp.283-296
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    • 2012
  • Allergic contact dermatitis (ACD) is an inflammatory skin disease and regarded as a prototype of T-cell mediated delayed-type hypersensitivity reactions. Poly-${\gamma}$-glutamic acid (PGA) is a biodegradable polymer that is produced by Bacillus subtilis. This study was performed to assess the effects of PGA in a canine model of ACD. ACD was induced on the back of dogs induced by sensitization and repeated application by 2,4-dinitro-1-chlorobenzene (DNCB). Topical treatment of PGA was applied once a day for 12 days and skin biophysical parameters including transepidermal water loss (TEWL), skin hydration, skin pH, skin thickness and erythema index, were measured every two days during experimental periods. Histopathology and immunohistochemistry were performed to evaluate the antiinflammatory effect. In skin biophysical parameters, TEWL, skin hydration, skin thickness and erythema index were significantly increased, with a maximum increase appeared on day 2 (p < 0.05). On the other hand, skin pH was significantly decreased, with a maximum decrease appeared on day 2 (p < 0.01). After the completion of PGA treatment, skin biophysical parameters were significantly reached those of baseline in a time-dependent manner (p < 0.05). In histopathology, marked increases of epidermal thicknesses were induced after DNCB challenge with numerous inflammatory cell infiltrations and edematous changes, decreases of connective tissue occupied regions in dermis. In addition, marked increases of cytokine - tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$)-immunoreactivities in the dermis and of apoptotic markers - caspase-3 and PARP-immunoreactivities in the epidermis were observed in DNCB-PBS control as compared with intact control, respectively (p < 0.01). It means, the ACD and related apoptotic changes were induced by DNCB in the present study. However, these ACD induced by DNCB and related apoptosis in epidermis were significantly inhibited by treatment of PGA treated skin, the decreases of infiltrated inflammatory cells and related decreases of pro-inflammatory cytokine immunoreactivities were also observed (p < 0.01). Based on these findings, PGA may have anti-inflammatory and alleviatory effects in the allergic contact dermatitis.

Protective Effects of New Herbal Composition (MH-30) against Radiation Injuries in Hematopoietic and Self-Renewal Tissues (생약조성물(MH-30)의 면역조혈계 및 재생조직 방사선 손상에 대한 방호 효과)

  • Jung, Uhee;Park, Hae-Ran;Lee, Ho-Yong;Baek, Ga-Young;Jo, Sung-Kee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.7
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    • pp.948-957
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    • 2016
  • We previously developed an herbal composition (HemoHIM) based on the water extracts of Angelica gigas radix, Cnidium officinale rhizoma, and Paeonia japonica radix to protect and recover hematopoietic and intestinal tissues against radiation injuries. In this study, to develop a composition with improved activities based on enhanced fat-soluble polyphenol contents, we prepared a new herbal composition, MH-30, from the above three herbs by 30% ethanol extraction and hot water extraction. HPLC analysis of the ethanol fractions of MH-30 and HemoHIM revealed that MH-30 had higher contents of many fat-soluble polyphenol compounds than HemoHIM (8.7-fold increase for decursin), whereas contents of water-soluble polyphenol compounds showed little differences between the two compositions. Then, we evaluated MH-30 and HemoHIM for their in vitro antioxidant and immune cell-stimulating activities as well as in vivo protective effects against radiation injuries in hematopoietic and self-renewal tissues. In antioxidant activity assays, MH-30 showed higher hydroxyl radical scavenging activity than HemoHIM (1.4- to 1.9-fold for compositions and 2.3- to 4.5-fold for ethanol fractions). On the other hand, MH-30 and HemoHIM exhibited similar immune cell-stimulating activities as measured by in vitro lymphocyte proliferation. MH-30 increased endogenous spleen colony formation, decreased bone marrow cell apoptosis, and enhanced survival of intestinal crypts in irradiated mice, demonstrating effective protection of MH-30 against radiation-induced injuries in hematopoietic and self-renewal tissues. The 30-day survival rate of lethally irradiated mice, a comprehensive index for radioprotective efficacy, was also elevated by MH-30. Noticeably, MH-30 showed higher protective effects than HemoHIM in all mouse experiments. These results demonstrate that MH-30 can protect hematopoietic and self-renewal tissues against radiation injuries more effectively than HemoHIM. Therefore, MH-30 can be a good candidate to reduce radiation injuries in hematopoietic and self-renewal tissues incurred by radiation accidents or cancer radiation therapy.

Effect of Stem Cell-Derived Conditioned Medium on the In Vitro Maturation and Embryonic Development of Parthenogenetic Embryos in Pigs (Stem Cell-Derived Conditioned Medium 첨가가 돼지난자의 체외성숙 및 단위발생란의 초기배 발육에 미치는 영향)

  • Kwon, Dae-Jin;Hwang, In-Sul;Kwak, Tae-Uk;Oh, Keon Bong;Ock, Sun-A;Chung, Hak-Jae;Im, Gi-Sun;Hwang, Seongsoo
    • Reproductive and Developmental Biology
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    • v.39 no.3
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    • pp.89-95
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    • 2015
  • The addition of growth factors and cytokines to in vitro culture (IVC) media could affect embryo development and the quality of the resulting blastocysts. The present study was performed to investigate the effect of porcine induced pluripotent stem cell (piPSC)-culture conditioned medium (CM) on the in vitro maturation (IVM) and development of parthenogentic embryos (parthenotes) in pigs. Cumulus-oocyte complexes (COCs) or activated oocytes were cultured in IVM or IVC medium supplemented with 0 (control), 25, or 50% of stem cell medium (SM) or CM, respectively. The maturation rate of CM-25% group was significantly improved when compared with control group (p<0.05), but that was not different among SM or CM groups. Blastocyst formation rate was significantly higher in CM-25% group (29.2%) than that of control (20.7%), SM-50% (19.6%) and CM-50% (23.66%, p<0.05). Cell number and the apoptotic cell index in blastocysts was significantly lower in SM-25% than in CM-25% group (p<0.05). The embryo quality related genes, OCT4, KLF4, TERT and ZFP42, were significantly increased in CM-25% group compared with control (p<0.05). In conclusion, the addition of 25% of CM to IVM and IVC medium positively influences not only the developmental potential also quality of parthenotes in pig.