• Title/Summary/Keyword: In vitro cytotoxicity

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Ethanol extract of Allium fistulosum inhibits development of non-alcoholic fatty liver disease

  • Hwang, Jin-Taek;Shin, Eun Ju;Chung, Min-Yu;Park, Jae Ho;Chung, Sangwon;Choi, Hyo-Kyoung
    • Nutrition Research and Practice
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    • v.12 no.2
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    • pp.110-117
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    • 2018
  • BACKGROUND/OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) is a leading cause of chronic liver disease and is closely associated with metabolic syndrome. In the present study, we observed the effect of ethanol extract of Allium fistulosum (EAF) on NAFLD and have suggested the possibility of using EAF as a natural product for application in the development of a treatment for NAFLD. MATERIALS/METHODS: The preventive effect on hepatic lipid accumulation was estimated by using an oleic acid (OA)-induced NAFLD model in vitro and a Western diet (high-fat high-sucrose; WD)-induced obese mouse model. Animals were divided into three groups (n = 7): normal diet group (ND), WD group, and WD plus 1% EAF group. RESULTS: EAF reduced OA-stimulated lipid accumulation in HepG2 cells in the absence of cellular cytotoxicity and significantly blocked transcriptional activation of sterol regulatory element-binding protein 1 and fatty acid synthase genes. Subsequently, we investigated these effects in vivo in mice fed either ND or WD in the presence or absence of EAF supplementation. In comparison to the ND controls, the WD-fed mice exhibited increases in body weight, liver weight, epididymal fat weight, and accumulation of fat in hepatocytes, and these effects were significantly attenuated by EAF supplementation. CONCLUSIONS: Allium fistulosum attenuates the development of NAFLD, and EAF elicits anti-lipogenic activity in liver. Therefore, EAF represents a promising candidate for use in the development of novel therapeutic drugs or drug combinations for the prevention and treatment of NAFLD.

In vivo and In vitro hair growth promotion effects of extract from Glycine soja Siebold et Zucc

  • Yang, Jae Chan;Kim, Bo Ae
    • Journal of Applied Biological Chemistry
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    • v.59 no.2
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    • pp.137-143
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    • 2016
  • Hair is a dermal adjunctive organ that protects the body from external physical and chemical stimuli; hair undergoes anagen, catagen, and telogen phases, with hair-loss occurring during the telogen phase. Alopecia is a condition wherein a person undergoes hair-loss far exceeding the normal amount, owing to diverse external factors. Wild beans are rich in isoflavone and amino acids known to prevent hair-loss; compared to cultivated beans, many wild bean species have higher protein content. This study aimed to develop a hair growth promoting solution, with superior hair growth promoting effects and fewer side effects, using naturally obtained Glycine soja Siebold et Zucc (GSSZ) extracts. Seven-week-old C57BL/6N male mice were classified into different experimental groups. Hair growth was observed in GSSZ-treated mice, and compared against that seen in 3 % minoxidil (MXD, positive control)-treated mice. Visual observations revealed a greater reduction in hair-loss in MXD and GSSZ application groups, compared to that in TXN group (hair loss induction using 1 % testosterone). Evaluation using an image analysis software revealed that compared to the positive control, TXN + GSSZ group showed the highest hair growth. TXN + MXD and control groups exhibited similar follicular cell growth, while the hair growth promotion patterns were similar in the negative control (normal), TXN + GSSZ, and TXN groups, as observed via histological analysis. GSSZ did not induce cytotoxicity (even at 2 mg/mL) in keratinocytes and dermal papilla cells; alternately, dermal papilla cell proliferation was activated in a (GSSZ) concentration-dependent manner. Therefore, the GSSZ extract promoted hair growth and increased hair growth-related cell activity, and could therefore be utilized in alopecia treatment.

Chemosensitizing effect and mechanism of imperatorin on the anti-tumor activity of doxorubicin in tumor cells and transplantation tumor model

  • Liang, Xin-li;Ji, Miao-miao;Liao, Zheng-gen;Zhao, Guo-wei;Tang, Xi-lan;Dong, Wei
    • The Korean Journal of Physiology and Pharmacology
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    • v.26 no.3
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    • pp.145-155
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    • 2022
  • Multidrug resistance of tumors has been a severe obstacle to the success of cancer chemotherapy. The study wants to investigate the reversal effects of imperatorin (IMP) on doxorubicin (DOX) resistance in K562/DOX leukemia cells, A2780/Taxol cells and in NOD/SCID mice, to explore the possible molecular mechanisms. K562/DOX and A2780/Taxol cells were treated with various concentrations of DOX and Taol with or without different concentrations of IMP, respectively. K562/DOX xenograft model was used to assess anti-tumor effect of IMP combined with DOX. MTT assay, Rhodamine 123 efflux assay, RT-PCR, and Western blot analysis were determined in vivo and in vitro. Results showed that IMP significantly enhanced the cytotoxicity of DOX and Taxol toward corresponding resistance cells. In vivo results illustrated both the tumor volume and tumor weight were significantly decreased after 2-week treatment with IMP combined with DOX compared to the DOX alone group. Western blotting and RT-PCR analyses indicated that IMP downregulated the expression of P-gp in K562/DOX xenograft tumors in NOD/SCID mice. We also evaluated glycolysis and glutamine metabolism in K562/DOX cells by measuring glucose consumption and lactate production. The results revealed that IMP could significantly reduce the glucose consumption and lactate production of K562/DOX cells. Furthermore, IMP could also remarkably repress the glutamine consumption, α-KG and ATP production of K562/DOX cells. Thus, IMP may sensitize K562/DOX cells to DOX and enhance the antitumor effect of DOX in K562/DOX xenograft tumors in NOD/SCID mice. IMP may be an adjuvant therapy to mitigate the multidrug resistance in leukemia chemotherapy.

Screening test for Dendropanax morbifera Leveille extracts: in vitro comparison to ox-LDL-induced lipid accumulation, ethanol-induced fatty liver and HMG-CoA reductase inhibition (황칠나무 추출물의 고지혈증 완화 효과 스크리닝)

  • Youn, Ji Sun;Kim, Min Seo;Na, Hye Jin;Jung, Hae Rim;Song, Chang Khil;Kang, So Young;Kim, Ji Yeon
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.1-8
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    • 2018
  • The objective of this study was to compare the antihyperlipidemic effects of different Dendropanax morbifera leaf extracts in vitro. The extracts differed in terms of specimen age, harvesting season, and extraction method. RAW 264.7 cells were pretreated with these extracts and stimulated by oxidized low-density lipoprotein. Ethanol was used to induce toxicity in HepG2 cells. Cellular lipid accumulation was quantified using oil red O staining in both these cells. The extracts were evaluated for their inhibitory effects on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. RAW 264.7 cells treated with the 60% ethanol extract of an 8-year-old specimen harvested in November exhibited the lowest lipid accumulation. The 30% ethanol extract of a 5-year-old specimen harvested in May exhibited the greatest protection from ethanol-induced cytotoxicity in HepG2 cells. The hot water extract of an 8-year-old specimen harvested in May showed the greatest inhibition of HMG-CoA reductase. These results showed that D. morbifera extracts prepared from leaves that are harvested in May possess the highest antihyperlipidemic effects.

Rare ginsenoside Ia synthesized from F1 by cloning and overexpression of the UDP-glycosyltransferase gene from Bacillus subtilis: synthesis, characterization, and in vitro melanogenesis inhibition activity in BL6B16 cells

  • Wang, Dan-Dan;Jin, Yan;Wang, Chao;Kim, Yeon-Ju;Perez, Zuly Elizabeth Jimenez;Baek, Nam In;Mathiyalagan, Ramya;Markus, Josua;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.42-49
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    • 2018
  • Background: Ginsenoside F1 has been described to possess skin-whitening effects on humans. We aimed to synthesize a new ginsenoside derivative from F1 and investigate its cytotoxicity and melanogenesis inhibitory activity in B16BL6 cells using recombinant glycosyltransferase enzyme. Glycosylation has the advantage of synthesizing rare chemical compounds from common compounds with great ease. Methods: UDP-glycosyltransferase (BSGT1) gene from Bacillus subtilis was selected for cloning. The recombinant glycosyltransferase enzyme was purified, characterized, and utilized to enzymatically transform F1 into its derivative. The new product was characterized by NMR techniques and evaluated by MTT, melanin count, and tyrosinase inhibition assay. Results: The new derivative was identified as (20S)-$3{\beta},6{\alpha},12{\beta}$,20-tetrahydroxydammar-24-ene-20-O-${\beta}$-D-glucopyranosyl-3-O-${\beta}$-D-glucopyranoside(ginsenoside Ia), which possesses an additional glucose linked into the C-3 position of substrate F1. Ia had been previously reported; however, no in vitro biological activity was further examined. This study focused on the mass production of arduous ginsenoside Ia from accessible F1 and its inhibitory effect of melanogenesis in B16BL6 cells. Ia showed greater inhibition of melanin and tyrosinase at $100{\mu}mol/L$ than F1 and arbutin. These results suggested that Ia decreased cellular melanin synthesis in B16BL6 cells through downregulation of tyrosinase activity. Conclusion: To our knowledge, this is the first study to report on the mass production of rare ginsenoside Ia from F1 using recombinant UDP-glycosyltransferase isolated from B. subtillis and its superior melanogenesis inhibitory activity in B16BL6 cells as compared to its precursor. In brief, ginsenoside Ia can be applied for further study in cosmetics.

Study on the development of polycaprolacton silica nanohybrid for bone substitutes (폴리카프로락톤 실리카 나노 복합체를 이용한 골이식대체재 개발에 관한 연구)

  • Jung, Keu-sik;Lim, Sung-Bin;Chung, Chin-Hyung;Hong, Ki-Seok;Kim, Jong-Yeo
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.425-448
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    • 2004
  • A bioactive and degradable poly(epsilon -caprolactone)/silica nanohybrid(PSH) was synthesized for the application as a bone substitute. PSH was manufactured by using silica and polycaprolacton. PSH was manufactured in some composition after low crystaline apatite had been formed in simulated body fluid and, was used this study. The safety of the PSH was established by test of acute, and subacute toxicity, sensitization cytotoxicity and sterility. In order to assess activity of osteoblast, the test for attaching osteoblast, proliferation test for osteoblast, differentiating gene expression test are performed in vitro. And bone substitutes were grafted in rabbit's calvarium, during 8 weeks for testing efficacy of bone substitutes. Degree of osteogenesis and absorption of substitutes were evaluated in microscopic level. In result, it was not appeared that acute and subacute toxicity, sensitization in intradermal induction phase, topical induction phase and challenge phase. It was shown that the test can not inhibit cell proliferation. adversely, it had some ability to accelerate cell proliferation. The result of sterility test described bacterial growth was not detected in most test tube. The attaching and proliferation test of osteoblast had good results. In the result of differentiating gene expression test for osteoblast, cbfa1 and, alkaline phosphatase, osteocalcin and GAPDH were detected with mRNA analysis. In the PSH bone formation test, ostgeoblastic activity would be different as material constitution but it had good new bone formation ability except group #218. futhermore, some material had been absorbed within 8 weeks. Above studies, PSH had bio-compatibility with human body, new bone formation ability and accelerate osteoblastic activity. So it would be the efficient bone substitute material with bio-active and biodegradable.

Study on Activities of Antioxidant and Anticancer of Germanium-fortified Tricholoma matsutake Myceliumm (게르마늄 강화 송이균사체의 항산화 및 항암 활성에 관한 연구)

  • Kim, Hae-Ja;Kim, Wan-Gyeom;Cho, Hwa-Eun;Choi, Yun-Hee;Lee, Ki-Nam;Chong, Myong-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.473-479
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    • 2009
  • The purpose of this study was evaluated physiological activity effect of organic germanium in culture broth of germanium-fortified Tricholoma matsutake mycelium and germanium-fortified yeast. Proliferation Tricholoma matsutake mycelium and yeast was inhibited by addition of germanium. Contents of organic germanium in Tricholoma matsutake mycelium and yeast was increased in dose-dependent manner. And low concentration(1,000 ppm) of germanium in mycelium was almost changed organic germanium. In the result of antioxidant activity as SOD-like activity, contents of total polyphenol compound and electron donating ability, activity of germanium-fortified Tricholoma matsutake mycelium was higher than that of germanium-fortified yeast. To evaluate of antitumor effects in vitro, we examined nitric oxide production of Raw 264,7 cell and cytotoxicity of HT1080 cell by MTT assay. Nitric oxide production of germanium-fortified Tricholoma matsutake mycelium was shown low level in low concentration(1,000 ppm) than other groups. The anticancer effect of germanium-fortified Tricholoma matsutake mycelium on HT 1080 cell was indicated a strong inhibitory effect in low concentration(1,000 ppm). These results suggest that organic germanium in culture broth of germanium-fortified Tricholoma matsutake mycelium has valuable physiological activities as antioxidant and anticancer effect, and it was higher than that of germanium-fortified yeast.

Synthesis and Evaluation of Antitumor Activity of Novel 1,4-Naphthoquinone Derivatives (IV)

  • Kim Bok Hee;Yoo Jikang;Park Si-Hyun;Jung Jae-Kyung;Cho Hoon;Chung Yongseog
    • Archives of Pharmacal Research
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    • v.29 no.2
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    • pp.123-130
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    • 2006
  • 1,4-Naphthoquinones are widely distributed in nature and many clinically important antitumor drugs containing a quinone moiety, such as anthracyclines, mitoxantrones and saintopin, show excellent anticancer activity. In this study, 2- or 6-substituted 5,8-dimethoxy-1,4-naphthoquinone (DMNQ) and 5,8-dihydroxy-1,4-naphthoquinone (DHNQ) derivatives were synthesized, and their cytotoxic activity against L1210 and P388 cancer cells was examined. Their antitumor activity was also assessed in mice bearing S-180 cells in the peritoneal cavity. In comparison with the DMNQ derivatives, the DHNQ derivatives exhibited more potent bioactivities than the DMNQ derivatives against both L1210 and P388 cells in vitro and S-180 cells in vivo. The $ED_{50}$ values of the DHNQ derivatives against P388 cells were in the range of 0.18-1.81 ${\mu}g/mL$ whereas those of the DMNQ derivatives were in the range of 0.26-40.41 ${\mu}g/mL$. The T/C ($\%$) values of the DHNQ derivatives, 8, 17, 18, 19, and 20, were found to be comparable to or even better than that of adriamycin. It was also observed that the 2-substituted derivatives (8, 19, 20) showed better antitumor activity than the 6-substituted derivatives (7, 17, 18) in the mice bearing S-180 cells in the peritoneal cavity.

Antinociceptive, anti-inflammatory, and cytotoxic properties of Origanum vulgare essential oil, rich with β-caryophyllene and β-caryophyllene oxide

  • Moghrovyan, Armenuhi;Parseghyan, Lilya;Sevoyan, Gohar;Darbinyan, Anna;Sahakyan, Naira;Gaboyan, Monica;Karabekian, Zaruhi;Voskanyan, Armen
    • The Korean Journal of Pain
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    • v.35 no.2
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    • pp.140-151
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    • 2022
  • Background: Essential oils are of great interest for their analgesic and anti-inflammatory properties. We aimed to study the content of the essential oil of the Origanum vulgare of the Armenian highlands (OVA) in different periods of vegetation and to investigate its antinociceptive and anti-inflammatory effects in mice (in vivo) and cytotoxic action in cultured cells (in vitro). OVA essential oil was extracted from fresh plant material by hydro-distillation. Methods: For OVA essential oil contents determination the gas chromatography-mass spectrometry method was used. Formalin and hot plate tests and analysis of cell viability using the methyl-thiazolyl-tetrazolium (MTT) assay were used. Results: The maximal content of β-caryophyllene and β-caryophyllene oxide in OVA essential oil was revealed in the period of blossoming (8.18% and 13.36%, correspondently). In the formalin test, 4% OVA essential oil solution (3.5 mg/mouse) exerts significant antinociceptive and anti-inflammatory effects (P = 0.003). MTT assay shows approximately 60% cytotoxicity in HeLa and Vero cells for 2.0 µL/mL OVA essential oil in media. Conclusions: The wild oregano herb of Armenian highlands, harvested in the blossoming period, may be considered as a valuable source for developing pain-relieving preparations.

Study of the Antioxidant Effect of Ojayeonjong-hwan (오자연종환(五子衍宗丸) 추출물의 항산화 효과에 관한 연구)

  • Jung, Yu-jin;Park, Sang-eun;Hong, Sang-hoon
    • The Journal of Internal Korean Medicine
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    • v.43 no.3
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    • pp.344-362
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    • 2022
  • Objective: In this study, the antioxidant activity of Ojayeonjong-hwan extracts was compared, and the following results were obtained. Methods: For hydrothermal and ethanol extracts, DPPH free radical and ABTS cationic radical erasing activity and reducing power using the FRAP method were compared, and the association between the antioxidant power of each extract and total phenol content was investigated. Significant results were obtained through in vitro apoptosis analysis through FFITC staining, mitochondrial membrane potential analysis, and ROS level measurement using C2C12 myoblastoma. Results: 1. In a comparison of DPPH free radical and ABTS cationic radical scavenging activity, water, and 70% ethanol extracts of Ojayeonjong-hwan (WEO and EEO) showed superior radical scavenging ability. 2. In the results of reducing power using the FRAP method, WEO and EEO showed antioxidant activity, which was shown to be dependent on the total phenol content contained in the extracts. 3. In comparison to the protective effect against H2O2-induced oxidative stress in C2C12 myoblasts, water extracts had no significant effect, but 70% ethanol extracts inhibited H2O2-mediated cytotoxicity in a concentration-dependent manner. 4. The cytotoxic protective effect of EEO against oxidative stress in C2C12 myoblasts was correlated with its inhibitory effects on H2O2-induced apoptosis and cell-cycle arrest. 5. In H2O2-treated C2C12 myoblasts, the apoptosis inhibitory effects of EEO were associated with the suppression of mitochondrial dysfunction and DNA damage. 6. The protective effects of EEO against H2O2-induced oxidative stress in C2C12 myoblasts were directly related to the inhibition of ROS generation. Conclusions: Ojayeonjong-hwan extracts all have protective potential against oxidative stress.