• Title/Summary/Keyword: RINm5F cells

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High Glucose Potentiates the Alloxan-induced Cytotoxicity in Cultured Rat Insulinoma Cells (흰쥐 인슐린종세포에서 고농도 포도당의 Alloxan 독성 증강 효과)

  • 이병래;차종희;박재윤;고춘남;박평심
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.5
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    • pp.875-880
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    • 2000
  • Reactive oxygen species are produced under diabetic conditions and possibly cause various forms of tissue damage in patients with diabetes mellitus. The aim of this study was to examine the effects of high glucose on the alloxan-induced beta cell injury. The insulinoma (RINm5F) cells were clutured either with high glucose (22.2 mM) or normoglucose (5.6 mM) in RPMI 1460 media for 3 days. The SOD activities were determined by spectrophotometric assay and nitroblue tetrazolium (NBT) stain. The effects of high glucose on the cytotoxicity of alloxan were also investigated in RINm5F cells and the cells viability were determined by 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) methods. Results showed that the CuZn-SOD activity was decreased but Mn-SOD activity was increased significantly in RINm5F cells cultured with high glucose (22.2 mM) media. The cytotoxicity of alloxan was increased by high glucose compared with normoglucose in RINm5F cells. Diethyl-dithiocarbarmate (DDC), as inhibitor of CuZn-SOC, also potentiate the alloxan-induced cytotoxocity in RINm5F cells. These results suggest that, in RINm5F cells, short term culture with high glucose media decreases Cu-Zn-SOD activity and the decreased activity of CuZn-SOD many one of the causative factors of beta-cell injury induced by high glucose.

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Protective Effect of Coptidis Rhizoma on SNAP-Induced Cytotoxicity in Pancreatic RINm5F Cells (SNAP 유도성 RINm5F 세포 독성에 대한 황연 추출물의 방어효과)

  • 류도곤;권강범;양정예;김은경;김강산
    • The Journal of Korean Medicine
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    • v.24 no.2
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    • pp.159-165
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    • 2003
  • Objectives : Coptidis rhizoma (CR) is an oriental medicine that has been used in many traditional prescriptions against diabetes mellitus in Korea for centuries. Our purpose was to determine the protective effect and its action mechanism of CR on the cytotoxicity of pancreatic -cell line (RINm5F cell). Methods : In this experiment, we used methods such as MTT assay for detection of cytotoxicity, DNA fragmentation assay for detection of apoptotic cell death, LDH activity assay for detection of necrotic cell death, and measurement of $DiOC_{6}$ (3) retention for detection of mitochondrial membrane potential (MMP). Background : Nitric oxide (NO) is believed to playa key role in the process of pancreatic -cell destruction leading to insulin-dependent diabetes mellitus (IDDM). Results : Exposure of RINm5F cells to chemical NO donor such as S-nitroso-N-acetylpenicillamine (SNAP) induced cytotoxic events such as DNA fragmentation and lactate dehydrogenase (LDH) release into medium. However, pretreatment of RINm5F cells with CR extract ($10~50{\mu\textrm{g}}/ml$) for 3 hours prevented SNAP-induced DNA fragmentation and LDH release into medium through the inhibition of MMP disruption. Conclusions : These results suggest that CR may be a candidate for a therapeutic or preventing agent against IDDM.

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Characterization of Voltage-Sensitive Calcium Channels and Insulin Secretion and the effect of 4,4'-Dichlorobiphenyl in RINm5f cells

  • Lee, Ihn-Soon;Hur, Eun-Mi;Sungkwon Chung;Kim, Kyong-Tai
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.47-47
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    • 2001
  • Opening of $Ca^{2+}$ -channels represents the final common pathway for insulin secretion in pancreatic beta-cells and related cell lines. We investigated voltage-sensitive calcium channels (VSCCs) and insulin secretion in RINm5F, an insulinoma cell line derived from rat pancreatic beta-cells. Several types of VSCCs were identified in RINm5f cells: dihydropyridine-sensitive L-type, $\omega$-conotoxin GVIA-sensitive N-type, $\omega$-agatoxin IVA-sensitive P-type channels, and $\omega$-conotoxin MVIIC sensitive Q-type channels.(omitted)

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Tat-CIAPIN1 protein prevents against cytokine-induced cytotoxicity in pancreatic RINm5F β-cells

  • Yeo, Hyeon Ji;Shin, Min Jea;Kim, Dae Won;Kwon, Hyeok Yil;Eum, Won Sik;Choi, Soo Young
    • BMB Reports
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    • v.54 no.9
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    • pp.458-463
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    • 2021
  • Cytokines activate inflammatory signals and are major mediators in progressive β-cell damage, which leads to type 1 diabetes mellitus. We recently showed that the cell-permeable Tat-CIAPIN1 fusion protein inhibits neuronal cell death induced by oxidative stress. However, how the Tat-CIAPIN1 protein affects cytokine-induced β-cell damage has not been investigated yet. Thus, we assessed whether the Tat-CIAPIN1 protein can protect RINm5F β-cells against cytokine-induced cytotoxicity. In cytokine-exposed RINm5F β-cells, the transduced Tat-CIAPIN1 protein elevated cell survivals and reduced reactive oxygen species (ROS) and DNA fragmentation levels. The Tat-CIAPIN1 protein reduced mitogen-activated protein kinases (MAPKs) and NF-κB activation levels and elevated Bcl-2 protein, whereas Bax and cleaved Caspase-3 proteins were decreased by this fusion protein. Thus, the protection of RINm5F β-cells by the Tat-CIAPIN1 protein against cytokine-induced cytotoxicity can suggest that the Tat-CIAPIN1 protein might be used as a therapeutic inhibitor against RINm5F β-cell damage.

The protective effect of Halal food extract in pancreatic beta cell lines.

  • Kim, Seong-sun;Jin, Yu-Mi;Song, Young-Jae;AYE, AYE;Soh, Ju-Ryoun;Jeon, Yong-Deok;Jin, Jong-Sik
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.121-121
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    • 2019
  • In Islamic dietary guidelines, Halal foods are allowed as edible blessed food. Most foods were categorized within halal for Muslims. The main point of Halal food is that foods are clean in every process and based on Halal standard which might be different in each country. Most pancreatic ${\beta}$ cells synthetize, store, and release insulin. Specific molecular, functional as well as ultrastructural traits of pancreatic ${\beta}$ cells could control their insulin secretion properties and survival phentoype. Insulin-secreting pancreatic ${\beta}$-cells are essential regulators of mammalian metabolism. In addition, the pancreatic ${\beta}$ cell plays an important role in the pathogenesis of type 1 and type 2 diabetes as improving glucose homeostasis by preserving, expanding and improving the function of this key cell type. However, the pharmacological effect of halal food has not been unclear yet, especially food habit-dependent diabetes. The aim of the this study was to determine the preventive effect of Iran plants extract (Almond, Garlic, Cumin, Ginkgo biloba, Holy basil, Psyllium, Satureja khuzistanica, Fenugreek, Green tea, Ipomoea betatas, Blueberry) on RINm5F cells and MIN6 cells as pancreatic ${\beta}$ cell line. The cytotoxicity of the extracts of Iran plants on RINm5F cells and MIN6 cells were measured by using MTT assays. The preventive effects of Iran plant extracts were measured by WST-8 cell proliferation assay on streptozotocin (STZ)-induced cell death in MIN6 cells. In presented result showed that all extract of Iran plants (0.01-10mg/ml) did not show cytotoxicity in RINm5F cells and MIN6 cells. Among non-cytotoxic extract, the protective effects could be detect in high dose concentration. These results suggest that the extract of Iran plants may serve as a potential therapy for diabetes.

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Prediabetic In vitro Model in Pancreatic Beta Cells Induced by Streptozotocin (췌장 베타세포에서 스트렙토조토신으로 유도한 인슐린 의존형 당뇨병 실험 모델)

  • Lee, Ihn-Soon;Rhee, In-Ja;Kim, Kyong-Tai
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.260-267
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    • 1997
  • To establish prediabetes in vitro model concerning the etiology of IDDM(Insulin Dependent Diabetes Mellitus) in cellular level we have designed prediabetes in vitro models in pa ncreatic beta cells. HIT-T15, RINm5F and isolated rat islets were chosen as pancreatic beta cells, and streptozotocin (STZ) used as diabetogenic agent. Degree of beta cell destruction to establish prediabetic in vitro model was determined by cell proliferation and insulin release using thymidine uptake and radio immuno assay. When HIT-T15 and RINm5F cells were treated with STZ, the degree of cell deterioration was dependent upon the origin and passage number of beta cells, and in the case of isolated islets STZ showed the more sensitivity than above two beta cell lines. The concentration and exposure time of STZ treatment to establish prediabetes in vitro model in beta cell lines and isolated rat islets were 2 ~ 10mM, 30 min. and 1 ~ 5mM, 30 min., respectively.

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Effect of Palmiwon on the Streptozotocin induced Prediabetic Model in Panceratic Bita Cells (췌장베타세포에서 스트렙토초토신으로 유도한 당뇨병 실험 모델에 대한 팔미원의 영향)

  • 이인순;이인자
    • Biomolecules & Therapeutics
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    • v.6 no.4
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    • pp.371-377
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    • 1998
  • The aim of the present study was to investigate the effect of Palmiwon on the type 1-prediabetic models induced by streptozotocin (STZ) in RINm5F cells and HIT-T15 cells. Palmiwon increased the cell proliferation and insulin release when pre- and post-treated for the STZ-exposed pancreatic beta cells. The cell proliferation and insulin release of these beta cells were measured by $^3$H-thymidine uptake and RIA, respectively. We also analyzed nutrients such as sugars, fatty acid and amino acids and minerals containing in Palmiwon using by gas chromatography, amino acid analyzer and AA spectrometer, respectively. Palmiwon seems to have protective and recovery properties on the prediabetic model in cellular level, which were ascribe to various nutrients and minerals containing in Palmiwon. From these results, it could be suggested that Palmiwon may be available as preventive and therapeutic prescription of type 1 diabetes mellitus.

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The Secretion Optimization of Oligopeptide with His-Pro Repeats in Bacillus subtilis and Its Anti-Diabetic Effects (고초균에서 His-Pro 반복서열을 갖는 Oligopeptide의 분비 최적화 및 항당뇨 효과)

  • Jeong, Seon Hwa;Choi, Jang Won
    • KSBB Journal
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    • v.32 no.1
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    • pp.71-82
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    • 2017
  • To verify anti-diabetic effect of oligopeptide with His-Pro repeats (mHP peptide), the oligopeptide was first secreted and optimized using the secretion vector, pRBAS with alkaline protease gene promoter and the signal sequence in Bacillus subtilis and directly the anti-diabetic effect of the mHP peptide was investigated in insulinoma cell, RINm5F cell line. The oligopeptide gene was obtained by annealing oligonucleotides with repeated His-Pro sequence and finally was constructed as 18 dipeptides (108 bp and 4.0 kDa) coding gene, named oligopeptide with His-Pro repeats (mHP peptide) to make cyclo(His-Pro) known to be anti-diabetic effects. The region encoding the oligopeptide gene was subcloned into the pRBAS secretion vector (E.coli-Bacillus shuttle vector) after PCR amplification using the designed primers including initiation and termination codons and His tag, named pRBAS-mHP (6.56 kb). To optimize secretion of the oligopeptide, various culture conditions were investigated in Bacillus subtilis LKS. As a result, the secreted oligopeptide was maximally measured (approximately $59.6{\mu}g/mL$) in 3 L batch culture and the highest secretion was achieved at $30^{\circ}C$, PY medium, and carbon sources (particularly barley and glycerol). In the RINm5F cells treated with 2 mM STZ, the oligopeptide treatment (0.1 mg/mL) restored the cell viability (10%) and reduced the nitric oxide (NO) generation (35%) and DNA fragmentation (90%). And also, insulin secretion level was increased to 17% higher than in STZ-treated RINm5F cells. These results suggest that the oligopeptide with His-Pro repeats could be a candidate material for anti-diabetic agent against STZ-induced diabetes.

In vitro Immunomodulating Effects of PALMIWON (팔미원의 in vitro 면역조절 작용)

  • Lee, Ihn-Soon;Rhee, In-Ja
    • YAKHAK HOEJI
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    • v.40 no.6
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    • pp.684-689
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    • 1996
  • PALMIWON is composed of 8 oriental herbs which has been known to show some pharmacological effects in kidney, blood vessels and immune systems, and used for the treatment of kid ney disease, hypertension, nervous disease and diabetic mellitus in the Orient for a long time. Based on our previous report that PALMIWON showed different effects on immune cells and ${\beta}$-cells, the immunoreactivity of ICSA (Islet Cell Surface Antibody) with ${\beta}$-cell (RINm5F) and the cell proliferation and function of interleukin-1${\beta}$ damaged ${\beta}$-cells in the presence of PALMIWON were examined. It was observed that PALMIWON significantly inhibited the immunoreactivity of ICSA with ${\beta}$-cell, and markedly increased cell proliferation and insulin release of interleukin-1${\beta}$ damaged ${\beta}$-cells.

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Protective Effect of Cordyceps sinensis Extract on Cytokine-induced Cytotoxicity of Pancreatic ${\beta}-cells$ (싸이토카인에 의한 췌장 ${\beta}$세포 독성에 대한 동충하초(冬蟲夏草) 추출물의 보호 효과)

  • Song, Je-Ho;Park, Byung-Hyun;Ryu, Do-Gon;Kwon, Kang-Beom
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.791-795
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    • 2008
  • In the present study, Cordyceps sinensis extract (CSE) was evaluated to determine if it could protect pancreatic ${\beta}$ cells against cytokine-induced cytotoxicity of RINm5F cells. Treatment of cells with cytokines resulted in a decrease of viability, which was caused by increase of nitric oxide (NO) production. CSE protected cytokine-mediated viability reduction in a concentration-dependent manner. Incubation with CSE also induced a significant suppression of cytokine-induced inducible nitric oxide synthase (iNOS) protein and NO production. The molecular mechanism by which CSE inhibited iNOS protein expression appeared to involve the inhibition of $NF-{\kappa}B$ activation. The cytokine-stimulated RIN cells showed increases in $NF-{\kappa}B$ binding activity compared to unstimulated cells. However, pretreatment with CSE inhibited cytokines-induced $NF-{\kappa}B$ activation in RINm5F cells.