• Title/Summary/Keyword: insulin cell

Search Result 604, Processing Time 0.026 seconds

Production of human insulin analogue using recombinant Escherichia coli

  • Lee, Ji-Seon;Park, Jin-Guk;Cho, Jung-Woo;Park, Sun-Ho;Nam, Doo-Hyun
    • 한국생물공학회:학술대회논문집
    • /
    • 2003.04a
    • /
    • pp.34-38
    • /
    • 2003
  • For the production of $B^{30}-homoserine$ insulin analog as a novel anti-diabetic drug, the fermentative study was attempted for the maximal gene expression of HTS-fused $B^{30}-homoserine$ insulin precursor in the recombinant Escherichia coli cells. In a batch fermentation, the maximal production of insulin precursor as much as 38.95 mg/L-h, which occupied more than 12.8% of total cell protein. was achieved when the gene expression was induced by 0.5 mM IPTG at the middle logarithmic growth phase. The HTS-fused $B^{30}-homoserine$ insulin precursor was recovered from a batch culture through the processes of cell harvest, collection of insoluble fraction after sonication and purification by nickel affinity column chromatography. The isolated insulin precursor was 14 mg/L with a recovery yield of 35.9% of expressed gene product. The insulin A and B chain mixture was recovered after the insulin precursor was subjected to CNBr cleavage and purified by nickel affinity column chromatography. The isolated insulin chains were then sulfitolyzed with sodium thiosulfat and sodium tetrathionate, and reconstituted to insulin analog with ${\beta}-mercaptoethanol$, followed by purification with CM-Sepharose C-25 column chromatography.

  • PDF

Effects of Insulin and IGFS on Growth and Functional Differentiation in Primary Cultured Rabbit Kidney Proximal Tubule Cells -Growth and membrane transport-

  • Han, Ho-Jae;Park, Kwon-Moo
    • The Korean Journal of Physiology
    • /
    • v.29 no.2
    • /
    • pp.191-202
    • /
    • 1995
  • The purpose of this study was to compare effects of insulin and IGFs on growth, apical membrane enzyme activities and membrane transport systems of primary cultured rabbit kidney proximal tubule cells. Results were as follows: 1. Insulin and IGF-I produced significant growth stimulatory effects at $5{\times}10^{-10}M.\;IGF-II(5×10^{-10}\;M)$ did not stimulate significant cell growth. 2. Insulin stimulated the phosphorylation of a 97 KD protein. It was difficult to determine whether this band represents insulin and/or the IGF-I receptor. 3. The activities of apical membrane enzymes (alkaline phosphatase, leucine aminopeptidase, and ${\gamma}-glutamyl \;transpeptidase)$ were observed to be diminished after the cells were placed in the culture environment. 4. The uptake of ${\alpha}-MG,$ Pi and Na was significantly increased in cells incubated with insulin or IGF-I, IGF-II had no effect on the uptake of these substrates. 5. Na-pump activity, as assayed by Rb uptake, was significantly increased in cells treated with insulin or IGFs. In conclusion, insulin and IGF-I exert stimulatory effects on growth and membrane transporter(glucose, Na, Pi, and Na-pump) activities in primary cultured rabbit kidney proximal tubule cells. IGF-II had no effect on cell growth and membrane transporter(glucose, Na and Pi) activities.

  • PDF

Electron microscopic study on the insulin-, glucagon-, somatostatin-, and pancreatic polypeptide secreting cells in Korean native goat (한국재래산양 췌장의 insulin, glucagon, somatostatin 및 pancreatic polypeptide 분비세포에 관한 전자현미경적 연구)

  • Lee, Heungshik S.;Lee, In-se;Kang, Tae-cheon;Won, Moo-ho;Yi, Seong-joon
    • Korean Journal of Veterinary Research
    • /
    • v.35 no.1
    • /
    • pp.55-65
    • /
    • 1995
  • Ultrastructures of pancreatic endocrine cells containing glucagon, insulin, somatosratin and pancreatic polypeptide were studied in the pancreas of the Korean native goat by immunohistochemical and elecron microscopy. Glucagon immunoreatctive cells were round or fusiform in shape and contained secretory granules of 200-260 nm in diameter. The secretory granules were high in electron density and had a halo between the limiting membrane and the central granule core. Insulin immunoreactive cells were round or oval in shape, and contained various sizes of secretory granules from 135 to 300 nm in diameter. The secretory granules were low or moderate electron density and had a variform halo. Somatostatin immunoreactive cells were elliptical or fusiform shape with cytoplasmic processes. They contained the secretory granules of 140-320 nm with moderate electron densities. Pancreatic polypeptide immunoreactive cells were elliptical or fusiform and contained small secretory granules with high electron densities. The secretory granules were 120-230 nm in diameter and the least in number.

  • PDF

In vitro Anti-diabetic Effects of Crude Extracts of Platycodi Radix (In vitro에서 길경 추출 분획물의 항당뇨 효과 조사)

  • Ko, Byoung-Seob;Kwon, Dae-Young;Hong, Sang-Mee;Park, Sun-Min
    • Korean Journal of Food Science and Technology
    • /
    • v.39 no.6
    • /
    • pp.701-707
    • /
    • 2007
  • Anti-diabetic effect of Platycodi radix (PR) extract fractions was determined if vitro by investigating insulin-like action, insulin sensitizing action, glucose-stimulated insulin secretion, gene expression related to ${\beta}-cell$ function and mass, and ${\alpha}$-glucoamylase suppressing action. Insulin-like activity was not promoted by the treatment of PR methanol factions in 373-L1 fibroblast. However, treatment with 0, 20 and 100% PR methanol fractions along with 1 ng/mL insulin increased insulin-stimulated glucose uptake in 373-L1 adipocytes. In addition, the treatment of 0% and 100% methanol fractions along with differentiation inducers significantly increased the differentiation of 373-L1 fibroblasts to adipocytes. These fractions may contain insulin sensitizer. The 20%, 80% and 100% methanol fractions enhanced glucose-stimulated insulin secretion in Min6 cells, insulin secreting cell line. This was related to the mechanism to promote glucose sensing and ${\beta}-cell$ proliferation, which was regulated by the induction of IRS-2, glucokinase and PDX-1 genes. As expected, 20, 80 and 100% methanol fractions increased mRNA levels of IRS-2, glucokinase and PDX-1 genes. However, PR fractions did not affect the ${\alpha}-glucoamylase$ activity in vitro. These data suggested that PR extract fractions have anti-diabetic actions through improving insulin sensitization, glucose-stimulated insulin secretion, and ${\beta}-cell$ proliferation. Therefore, PR extracts can be beneficial for anti-diabetic treatment in lean diabetic patients.

Relationship between Body Mass Index and Insulin Resistance and Beta Cell Function by Obesity Type in Obese Middle-Aged Men (비만 중년 남성의 비만 유형별 체질량지수와 인슐린저항성 및 베타세포 기능간의 관련성)

  • Shin, Kyung-A
    • Journal of the Korea Convergence Society
    • /
    • v.11 no.10
    • /
    • pp.155-162
    • /
    • 2020
  • This study wanted to confirm the relevance between body mass index(BMI) and insulin resistance and beta-cell function based on abdominal obesity in obese middle-aged men. This study targeted 797 obese middle-aged men who had undergone health checkups at general hospitals in Gyeonggi-do from January 2018 to June 2020. There were 327 in the group with abdominal obesity and 470 in the group without abdominal obesity. Glucose(p<0.001), HbA1c(p=0.003), insulin(p<0.001), HOMA-IR(p<0.001) was different between groups. BMI was a factor affecting insulin resistance and beta cell function regardless of the with or without of abdominal obesity. BMI was associated with the onset of disease of insulin resistance and beta cell functional degradation regardless of the with or without of abdominal obesity. Therefore, it is considered necessary to manage the indicators of the BMI through exercise programs and regular checkups for health management of middle-aged obese men.

NK cell-activating receptor NKp46 does not participate in the development of obesity-induced inflammation and insulin resistance

  • Gracia Nathalie;Beatriz Dal Santo Francisco Bonamichi;Jieun Kim;Jiwon Jeong;Haneul Kang;Emirrio Reinaldie Hartland;Eveline Eveline;Jongsoon Lee
    • Molecules and Cells
    • /
    • v.47 no.3
    • /
    • pp.100007.1-100007.11
    • /
    • 2024
  • Recent evidence establishes a pivotal role for obesity-induced inflammation in precipitating insulin resistance and type-2 diabetes. Central to this process is the proinflammatory M1 adipose-tissue macrophages (ATMs) in epididymal white adipose tissue (eWAT). Notably, natural killer (NK) cells are a crucial regulator of ATMs since their cytokines induce ATM recruitment and M1 polarization. The importance of NK cells is shown by the strong increase in NK-cell numbers in eWAT, and by studies showing that removing and expanding NK cells respectively improve and worsen obesity-induced insulin resistance. It has been suggested that NK cells are activated by unknown ligands on obesity-stressed adipocytes that bind to NKp46 (encoded by Ncr1), which is an activating NK-cell receptor. This was supported by a study showing that NKp46-knockout mice have improved obesity-induced inflammation/insulin resistance. We therefore planned to use the NKp46-knockout mice to further elucidate the molecular mechanism by which NKp46 mediates eWAT NK-cell activation in obesity. We confirmed that obesity increased eWAT NKp46+ NK-cell numbers and NKp46 expression in wild-type mice and that NKp46-knockout ablated these responses. Unexpectedly, however, NKp46-knockout mice demonstrated insulin resistance similar to wild-type mice, as shown by fasting blood glucose/insulin levels and glucose/insulin tolerance tests. Obesityinduced increases in eWAT ATM numbers and proinflammatory gene expression were also similar. Thus, contrary to previously published results, NKp46 does not regulate obesity-induced insulin resistance. It is therefore unclear whether NKp46 participates in the development of obesity-induced inflammation and insulin resistance. This should be considered when elucidating the obesity-mediated molecular mechanisms that activate NK cells.

MODULATION OF IRRADIATION-INDUCED CELL DEATH BY INSULIN-LIKE GROWTH FACTOR-II IN MC3T3 OSTEOBLASTS (Insulin-like growth factor-II가 방사선에 의한 MC3T3 조골세포의 세포사멸에 미치는 영향)

  • Park, Kyeong-Lok
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.33 no.6
    • /
    • pp.617-624
    • /
    • 2007
  • Insulin-like growth factor(IGF) is the most abundant growth factor in bone matrix. Recent studies have shown that it can sensitize apoptotic cell death of osteoblasts. Thus, this study investigated whether IGF-II aggravates irradiation-induced cell death of osteoblasts. Cultured MC3T3 osteoblasts were irradiated and IGF-II was added at the concentration of 50 ng/ml immediately after the irradiation. Cell viability was measured by MTT assay. Changes in cell death and cell cycle were analyzed by flow cytometry. The expression of proapoptotic gene bax and antiapoptotic gene bcl-2 was quantified by real time RT-PCR and Western blot. A dose of 30 Gy caused G2/M arrest and increased cell death through both necrosis and apoptosis, while irradiation from 4 to 10 Gy little affected cell cycle and death. IGF-II treatment reduced cell viability without stimulating cell proliferation and changing cell cycle. Combined treatment of IGF-II with irradiation decreased cell viability and proliferation and increased cell death along with G2/M arrest. These effects were not different from those of irradiation only. At transcriptional and protein levels, IGF-II treatment did not affect bax and bcl-2 expression, whereas irradiation increased the expression ofbax without changes in bcl-2. IGF-II in combination with irradiation showed similar findings. These results suggest that IGF-II could modulate apoptotic cell death through mechanisms other than an imbalance between bax and bcl-2 gene expression, although its effect was overridden by irradiation.

Clusterin(SGP-2) in the Salivary Glands of Insulin Injected Rats under Stress (스트레스에 의한 혈당변화가 타액선내 Clusterin 발현에 미치는 영향)

  • 김선호
    • Journal of Oral Medicine and Pain
    • /
    • v.23 no.4
    • /
    • pp.309-326
    • /
    • 1998
  • In general, the major causative factor of halitosis is thought to be a sulfated compounds. Clusterin, a sulfated glycoprotein-2(SGP-2), is frequently found in diabetic conditions and cold stress conditions. The same result is werum glucose level to diabeteic and cold stress conditions that founded Clusterin. Therefore, this study was performed to examine Clusterin in the slivary glands under stress conditions before insulin injection I.M. Fourty rats were diveded into 3 groups ; 1) 10 rats of gorup I were selected as a control 2) 15 rats beloning to group II were bathed in cold water for 30 seconds twice a day 3) 15 rats in group III received cold stress and injected I.M. with insulin. The rats were sacrifeced at day 0, 3, 5, 7 and 10 of the experiment and the submandibular glands and parotid galnds were removed. RNAs were purified from the salivary of the salivary glands were subjected to Hematoxillin-Eosin stainings and examined under the light microscope. The obtained results were as follows : 1. With immunohistochemistric method, in normal control goup, Clusterin was moderately stained in the intercalated ductal cell of the submandibualr glands, mild stained in the striated ductal cell of the submandibular glands, heavily stained on the cytoplasm of the intercalated ductal cell in the mucous submandibular glands nad slightrly stained in the intercalated ductal cell of the paroted gland, expressed negativity in the acina cell. 2. With immunohistochemistric method, Clusterin slightly increased in the acina cell of the submandibular glands under stress condition at 3 days after experement, moderately stained at 5 days after experiment so revealed positive response. And hearily in the intercalated ductal cell and mildly lin the acina celluar eytoplasm of the parotid glands under stress condition at 3 days experiment. 3. With immunohistochemistric method, no remarkable differences are found between the normal control group and stress conditioned group that insulin administration was performed before. 4. In the stressor-giving group, Clusterin mRNA was porminently expressed in submandibular gland after 5 days after experiment, in parotid gland after 3 days after experiment, performed in immunoelectrophoresis method. 5. In the insulin-injected nad stressor-giving group, Clusterin mRNA was not observed in all experimental submandibular and parotid gland, performed in immunoelectrophoresis method.

  • PDF

Effect of White, Taegeuk, and Red Ginseng Root Extracts on Insulin-Stimulated Glucose Uptake in Muscle Cells and Proliferation of β-cells

  • Cha, Ji-Young;Park, Eun-Young;Kim, Ha-Jung;Park, Sang-Un;Nam, Ki-Yeul;Choi, Jae-Eul;Jun, Hee-Sook
    • Journal of Ginseng Research
    • /
    • v.34 no.3
    • /
    • pp.192-197
    • /
    • 2010
  • Recent studies have indicated that $\beta$-cell dysfunction and insulin resistance are important factors in the development of type 2 diabetes. The present study investigated the effect of extracts from different parts of white, Taegeuk, and red ginseng root on insulin-stimulated glucose uptake in muscle cells and proliferation of $\beta$-cells. Extracts of the fine roots of Taegeuk ginseng significantly enhanced glucose uptake compared with the control. White ginseng lateral root extracts enhanced insulin-induced glucose uptake. Proliferation of $\beta$-cells was significantly increased by Taegeuk ginseng main and lateral root extracts and by red ginseng lateral and fine root extracts. In conclusion, different root parts of white, Taegeuk, and red ginseng differentially affect glucose uptake and pancreatic $\beta$-cell proliferation.