• Title/Summary/Keyword: in vitro experiments

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Effects of Oleo Gum Resin of Ferula assa-foetida L. on Senescence in Human Dermal Fibroblasts - Asafoetida reverses senescence in fibroblasts -

  • Moghadam, Farshad Homayouni;Mesbah-Ardakani, Mehrnaz;Nasr-Esfahani, Mohammad Hossein
    • Journal of Pharmacopuncture
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    • v.20 no.3
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    • pp.213-219
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    • 2017
  • Objectives: Based on data from Chinese and Indian traditional herbal medicines, gum resin of Ferula assa-foetida (sometimes referred to asafetida or asafoetida) has several therapeutic applications. The authors of various studies have claimed that asafetida has cytotoxic, antiulcer, anti-neoplasm, anti-cancer, and anti-oxidative effects. In present study, the anti-aging effect of asafetida on senescent human dermal fibroblasts was evaluated. Methods: Senescence was induced in in vitro cultured human dermal fibroblasts (HDFs) through exposure to $H_2O_2$, and the incidence of senescence was recognized by using cytochemical staining for the activity of ${\beta}$-galactosidase. Then, treatment with oleo gum resin of asafetida was started to evaluate its rejuvenating effect. The survival rate of fibroblasts was evaluated by using methyl tetrazolium bromide (MTT) assays. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blot assays were performed to evaluate the expressions of apoptotic and anti-apoptotic markers. Results: Our experiments show that asafetida in concentrations ranging from $5{\times}10^{-8}$ to $10^{-7}g/mL$ has revitalizing effects on senescent fibroblasts and significantly reduces the ${\beta}$-galactosidase activity in these cells (P < 0.05). Likewise, treatment at these concentrations increases the proliferation rate of normal fibroblasts (P < 0.05). However, at concentrations higher than $5{\times}10^{-7}g/mL$, asafetida is toxic for cells and induces cell death. Conclusion: The results of this study indicate that asafetida at low concentrations has a rejuvenating effect on senescent fibroblasts whereas at higher concentrations, it has the opposite effect of facilitating cellular apoptosis and death.

Analysis of an External Stimulator's Impact on the Heart (체외 전기자극기가 심장에 미치는 영향의 분석 및 평가)

  • Kim, Mun-Soo;Choe, Seung-Wook
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.11
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    • pp.1213-1217
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    • 2011
  • Electric stimulators are used for various treatments, such as for pain relief and to improve rehabilitation in and out of the hospital. However, if the stimulation pulse affects the patient's heart, it can cause critical cardiac disorders such as arrhythmia or ventricular fibrillation. As a result, it must be ensured that the transmission length of the stimulation pulse does not exceed the proper range in the design of an electric stimulator. Furthermore, every anticipated risk factor must be monitored in in-vitro and in-vivo experiments. A new stimulator was designed to supply continuous 0.001 J stimulation pulses at a rate of 60 pulses per second. To evaluate the safety of the new electric stimulator and to measure its energy transfer and pulse transmission length, we built a conduction model that was filled with saline and measured the electric field at various positions in response to real stimulations. In an animal experiment with two pigs, heart disorders were induced by applying electric stimulation to tissues near the heart. These heart disorders were different from the result obtained with 9 V DC stimulation.

Suppression of Bacterial Wilt in Tomato Plant Using Pseudomonas putida P84 (Pseudomonas putida P84 균주를 이용한 토마토 풋마름병의 억제)

  • Seo, Sang-Tae;Park, Jong-Han;Kim, Kyung-Hee;Lee, Sang-Hyun;Oh, Eun-Sung;Shin, Sang-Chul
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.32-36
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    • 2008
  • Bacterial wilt caused by Ralstonia solanacearum has become a severe problem on tomato in Korea and no effective control measures are available yet. Pseudomonas species play key roles for the biocontrol of many plant diseases especially in soil. A rhizobacterial population of 150 Pseudomonas strains, isolated from the rhizosphere soil of various plants grown at different sites, was screened for 2,4-diacetylphloroglucinol producing gene (PhlD) by PCR. Two strains (P83 and P84) among them were found to be phlD positive. When the isolates were analysed by 16S rDNA (Sensu Stricto), all isolates yielded amplified products of 1,018bp. Of the 150 isolates of Pseudomonas spp., a bacterial strain P. putida P84 isolated from tomato rhizosphere showed to suppress a wide range of phytopathogenic bacteria in vitro. The best source of carbon for P84 strain were glucose, arabinose, inositol and melibiose. In greenhouse experiments, P84 strain suppressed the development of bacterial wilt in tomato with a control value of 60%.

The Effect of Palmultang(八物湯) on the Ovarian Functions and Differential Gene Expression of Caspase-3, MAPK and MPG in Female Mice (팔물탕(八物湯)이 자성생쥐의 생식능력과 Caspase-3, MAPK 및 MPG 유전자 발현에 미치는 영향)

  • Joo, Jin-Man;Baek, Seung-Hee;Kim, Eun-Ha;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.3
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    • pp.91-110
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    • 2007
  • Purpose : These experiments were undertaken to evaluate the effect of administration of Palmultang on ovarian functions and differential gene expressions related cell viabilities caspase-3, MAPK and MPG in female mice. Materials and Methods : We administered the Palmultang to 6-week-old female ICR mice for 4, 8, or 12 days. The female mice were injected PMSG and hCG for ovarian hyperstimulation. And then recovered ovaries were minced and extracted mRNA and analyzed cell viability related gene expression. We chose the caspase-3 for cell apoptosis, MAPK and MPG genes for cell viability and DNA repair. To compare the differences, we set a control group treated with plain water at the same volume by the same way. Results : In case of administration of Palmultang, the mean number of total ovulated oocytes and the number of morphologically normal oocytes increased significantly compared to a control group. We were also examined the embryonic developmental competence in vitro. The administration of Palmultang in a concentration with 10 and 100 mg/ml were beneficial effect of embryonic development in preimplantation period. The administration of Palmultang play a role of prevention of cell apoptosis and DNA damages and also increased cell proliferation resulted in ovarian functions. Conclusion : From our results suggested that the medication of Palmultang has beneficial effect on reproductive functions of female mice via prevention of cell apoptosis and DNA damaging and promotion of cell proliferation.

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Probiotic Characteristics of Lactobacillus brevis KT38-3 Isolated from an Artisanal Tulum Cheese

  • Hacioglu, Seda;Kunduhoglu, Buket
    • Food Science of Animal Resources
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    • v.41 no.6
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    • pp.967-982
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    • 2021
  • Probiotics are living microorganisms that, when administered in adequate amounts, provide a health benefit to the host and are considered safe. Most probiotic strains that are beneficial to human health are included in the "Lactic acid bacteria" (LAB) group. The positive effects of probiotic bacteria on the host's health are species-specific and even strain-specific. Therefore, evaluating the probiotic potential of both wild and novel strains is essential. In this study, the probiotic characteristics of Lactobacillus brevis KT38-3 were determined. The strain identification was achieved by 16S rRNA sequencing. API-ZYM test kits were used to determine the enzymatic capacity of the strain. L. brevis KT38-3 was able to survive in conditions with a broad pH range (pH 2-7), range of bile salts (0.3%-1%) and conditions that simulated gastric juice and intestinal juice. The percentage of autoaggregation (59.4%), coaggregation with E. coli O157:H7 (37.4%) and hydrophobicity were determined to be 51.1%, 47.4%, and 52.7%, respectively. L. brevis KT38-3 produced β-galactosidase enzymes and was able ferment lactose. In addition, this strain was capable of producing antimicrobial peptides against the bacteria tested, including methicillin and/or vancomycin-resistant bacteria. The cell-free supernatants of the strain had high antioxidant activities (DPPH: 54.9% and ABTS: 48.7%). Therefore, considering these many essential in vitro probiotic properties, L. brevis KT38-3 has the potential to be used as a probiotic supplement. Supporting these findings with in vivo experiments to evaluate the potential health benefits will be the subject of our future work.

GATA2-Mediated Transcriptional Activation of Notch3 Promotes Pancreatic Cancer Liver Metastasis

  • Lin, Heng;Hu Peng;Zhang, Hongyu;Deng, Yong;Yang, Zhiqing;Zhang, Leida
    • Molecules and Cells
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    • v.45 no.5
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    • pp.329-342
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    • 2022
  • The liver is the predominant metastatic site for pancreatic cancer. However, the factors that determine the liver metastasis and the specific molecular mechanisms are still unclear. In this study, we used human pancreatic cancer cell line Hs766T to establish Hs766T-L3, a subline of Hs766T with stable liver metastatic ability. We performed RNA sequencing of Hs766T-L3 and its parental cell line Hs766T, and revealed huge differences in gene expression patterns and pathway activation between these two cell lines. We correlated the difference in pathway activation with the expression of the four core transcriptional factors including STAT1, NR2F2, GATA2, and SMAD4. Using the TCGA database, we examined the relative expression of these transcription factors (TFs) in pan-cancer and their relationship with the prognosis of the pancreatic cancer. Among these TFs, we considered GATA2 is closely involved in tumor metastasis and may serve as a potential metastatic driver. Further in vitro and in vivo experiments confirmed that GATA2-mediated transcriptional activation of Notch3 promotes the liver metastasis of Hs766T-L3, and knockdown of either GATA2 or Notch3 reduces the metastatic ability of Hs766T-L3. Therefore, we claim that GATA2 may serve as a metastatic driver of pancreatic cancer and a potential therapeutic target to treat liver metastasis of pancreatic cancer.

Monitoring Cellular Immune Responses after Consumption of Selected Probiotics in Immunocompromised Mice

  • Kang, Seok-Jin;Yang, Jun;Lee, Na-Young;Lee, Chang-Hee;Park, In-Byung;Park, Si-Won;Lee, Hyeon Jeong;Park, Hae-Won;Yun, Hyun Sun;Chun, Taehoon
    • Food Science of Animal Resources
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    • v.42 no.5
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    • pp.903-914
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    • 2022
  • Probiotics are currently considered as one of tools to modulate immune responses under specific clinical conditions. The purpose of this study was to evaluate whether oral administration of three different probiotics (Lactiplantibacillus plantarum CJLP243, CJW55-10, and CJLP475) could evoke a cell-mediated immunity in immunodeficient mice. Before conducting in vivo experiments, we examined the in vitro potency of these probiotics for macrophage activation. After co-culture with these probiotics, bone marrow derived macrophages (BMDMs) produced significant amounts of proinflammatory cytokines including interleukin-6 (IL-6), IL-12, and tumor necrosis factor-α (TNF-α). Levels of inducible nitric oxide synthase (inos) and co-stimulatory molecules (CD80 and CD86) were also upregulated in BMDMs after treatment with some of these probiotics. To establish an immunocompromised animal model, we intraperitoneally injected mice with cyclophosphamide on day 0 and again on day 2. Starting day 3, we orally administered probiotics every day for the last 15 d. After sacrificing experimental mice on day 18, splenocytes were isolated and co-cultured with these probiotics for 3 d to measure levels of several cytokines and immune cell proliferation. Results clearly indicated that the consumption of all three probiotic strains promoted secretion of interferon-γ (IFN-γ), IL-1β, IL-6, IL-12, and TNF-α. NK cell cytotoxicity and proliferation of immune cells were also increased. Taken together, our data strongly suggest that consumption of some probiotics might induce cell-mediated immune responses in immunocompromised mice.

Effect of Evodiae Fructus on the ovarian function and gene expression of caspase-3, MAP kinase and MPG in female mice (오수유 투여가 자성생쥐의 생식능력과 caspase-3, MAPK 및 MPG유전자 발현에 미치는 영향)

  • Lee, Ja-Young;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.2
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    • pp.60-78
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    • 2009
  • Purpose: These experiments were undertaken to evaluate the effect of administration of Evodiae Fructus on ovarian functions and differential gene expressions related caspase-3, MAPK and MPG in female mice. Methods: We administered the Evodiae Fructus to 6-week-old female ICR mice for 4, 8, or 12 days. With different concentration of Evodiae Fructus, the female mice were injected PMSG and hCG for ovarian hyperstimulation. The mice divided into 3 groups for each experiment. We chose the caspase-3 for cell apoptosis, MAPK and MPG genes for cell viability and DNA repair. Results: In case of 4, 8, 12 day of Evodiae Fructus, we were examined the mean number of total ovulated oocytes and the number of morphologically normal oocytes. We were also examined the embryonic developmental competence in vitro. In addition we were also examined the differential expression of cell viability related genes, caspase-3, MAPK and MPG according to concentration and duration of Evodiae Fructus administration. MPG gene expressions for cell viability and DNA repaie were increased in dose dependent manner than that of control group in 4-day administration group. Conclusion: It is suggested that the medication of Evodiae Fructus has beneficial effect on reproductive functions of female mice via promotion of cell proliferation.

Ginsenoside Rg3 ameliorates myocardial glucose metabolism and insulin resistance via activating the AMPK signaling pathway

  • Ni, Jingyu;Liu, Zhihao;Jiang, Miaomiao;Li, Lan;Deng, Jie;Wang, Xiaodan;Su, Jing;Zhu, Yan;He, Feng;Mao, Jingyuan;Gao, Xiumei;Fan, Guanwei
    • Journal of Ginseng Research
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    • v.46 no.2
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    • pp.235-247
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    • 2022
  • Background: Ginsenoside Rg3 is one of the main active ingredients in ginseng. Here, we aimed to confirm its protective effect on the heart function in transverse aortic coarctation (TAC)-induced heart failure mice and explore the potential molecular mechanisms involved. Methods: The effects of ginsenoside Rg3 on heart and mitochondrial function were investigated by treating TAC-induced heart failure in mice. The mechanism of ginsenoside Rg3 for improving heart and mitochondrial function in mice with heart failure was predicted through integrative analysis of the proteome and plasma metabolome. Glucose uptake and myocardial insulin sensitivity were evaluated using micro-positron emission tomography. The effect of ginsenoside Rg3 on myocardial insulin sensitivity was clarified by combining in vivo animal experiments and in vitro cell experiments. Results: Treatment of TAC-induced mouse models with ginsenoside Rg3 significantly improved heart function and protected mitochondrial structure and function. Fusion of metabolomics, proteomics, and targeted metabolomics data showed that Rg3 regulated the glycolysis process, and Rg3 not only regulated glucose uptake but also improve myocardial insulin resistance. The molecular mechanism of ginsenoside Rg3 regulation of glucose metabolism was determined by exploring the interaction pathways of AMPK, insulin resistance, and glucose metabolism. The effect of ginsenoside Rg3 on the promotion of glucose uptake in IR-H9c2 cells by AMPK activation was dependent on the insulin signaling pathway. Conclusions: Ginsenoside Rg3 modulates glucose metabolism and significantly ameliorates insulin resistance through activation of the AMPK pathway.

Study on Anti-Skin Aging Effect of Sanguisorba officinalis L. (지유(地楡)의 피부 노화(老化)에 대한 연구(硏究))

  • Kim, Kyoung-Shin;Tak, Dong-Yul;Kim, Byoung-Soo;Kang, Jung Soo
    • Journal of Haehwa Medicine
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    • v.21 no.2
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    • pp.63-72
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    • 2013
  • To develop a new anti-skin aging cosmetics or functional foods by using antioxidative activity and collagenase inhibitor, a potent collagenase or elastase inhibitor was screened from various extracts of medicinal plants and its optimal extraction condition was investigated. And antioxidative activity, antimicrobial activity and inhibitory of effect against collagenase activity were investigated. In the these results, we selected the Sanguisorba (Sanguisorba officinalis L.) that presents a potential biological activities. Sanguisorba which is very rich in triterpenoid saponin and tannins was recently reported its anti-oxidant activities and phytoestogenic activities in vivo test and many clinical studies. The experiments were carried out in vitro to determine anti-oxidant activities of Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, Elastase and collagenase activity assay. It show that the Sanguisorba extracts have the most significant anti-oxidant on free radical scavenging activity assay, and also inhibited significantly activities of elastase, collagenase. Further, Sanguisorba extracts are activated Type I collagen protein expression in CCD-986sk cells. These result suggest that the Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, elastase and collagenase activity assay, Type I collagen protein expression in CCD-986sk cells effected could be developed cosmetic ingredients for anti-aging.