• Title/Summary/Keyword: IGF-l

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Glucosamine Hydrochloride and N-Acetylglucosamine Influence the Response of Bovine Chondrocytes to TGF-β3 and IGF in Monolayer and Three-Dimensional Tissue Culture

  • Pizzolatti, Andre Luiz A.;Gaudig, Florian;Seitz, Daniel;Roesler, Carlos R.M.;Salmoria, Gean Vitor
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.781-791
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    • 2018
  • BACKGROUND: Glucosamine hydrochloride (GlcN HCl) has been shown to inhibit cell growth and matrix synthesis, but not with N-acetyl-glucosamine (GlcNAc) supplementation. This effect might be related to an inhibition of critical growth factors (GF), or to a different metabolization of the two glucosamine derivatives. The aim of the present study was to evaluate the synergy between GlcN HCl, GlcNAc, and GF on proliferation and cartilage matrix synthesis. METHOD: Bovine chondrocytes were cultivated in monolayers for 48 h and in three-dimensional (3D) chitosan scaffolds for 30 days in perfusion bioreactors. Serum-free (SF) medium was supplemented with either growth factors (GF) $TGF-{\beta}$ ($5ng\;mL^{-1}$) and IGF-I ($10ng\;mL^{-1}$), GlcN HCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF + GlcN HCl; (c) SF + GlcNAc; (d) SF + GF; (e) SF + GF + GlcN HCl; and (f) SF + GF + GlcNAc. Cell proliferation, proteoglycan, collagen I (COL1), and collagen II (COL2) synthesis were evaluated. RESULTS: The two glucosamines showed opposite effects in monolayer culture: GlcN HCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcN HCl added to GF stimulated cell proliferation more than when compared to GF only, but the proteoglycan synthesis was smaller than GF. However, GlcNAc added to GF improved the cell proliferation and proteoglycan synthesis more than when compared to GF and GF/GlcN HCl. The synthesis of COL1 and COL2 was observed in all groups containing GF. CONCLUSION: GlcN HCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.

Increased interleukin-6 and TP53 levels in rotator cuff tendon repair patients with hypercholesterolemia

  • Jong Pil Yoon;Seung Gi Min;Jin-Hyun Choi;Hyun Joo Lee;Kyeong Hyeon Park;Sung Hyuk Yoon;Seong Soo Kim;Seok Won Chung;Hun-Min Kim;Dong Hyun Kim
    • Clinics in Shoulder and Elbow
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    • v.25 no.4
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    • pp.296-303
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    • 2022
  • Background: A previous study reported that hyperlipidemia increases the incidence of tears in the rotator cuff tendon and affects healing after repair. The aim of our study was to compare the gene and protein expression of torn rotator cuff tendons in patients both with and without hypercholesterolemia. Methods: Thirty patients who provided rotator cuff tendon samples were classified into either a non-hypercholesterolemia group (n=19, serum total cholesterol [TC] <200 mg/dL) and hypercholesterolemia group (n=11, serum TC ≥240 mg/dL) based on their concentrations of serum TC. The expression of various genes of interest, including COL1A1, IGF1, IL-6, MMP2, MMP3, MMP9, MMP13, TNMD, and TP53, was analyzed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). In addition, Western blot analysis was performed on the proteins encoded by interleukin (IL)-6 and TP53 that showed significantly different expression levels in real-time qRT-PCR. Results: Except for IGF1, the gene expression levels of IL-6, MMP2, MMP9, and TP53 were significantly higher in the hypercholesterolemic group than in the non-hypercholesterolemia group. Western blot analysis confirmed significantly higher protein levels of IL-6 and TP53 in the hypercholesterolemic group (p<0.05). Conclusions: We observed an increase in inflammatory cytokine and matrix metalloproteinase (MMP) levels in hypercholesterolemic patients with rotator cuff tears. Increased levels of IL-6 and TP53 were observed at both the mRNA and protein levels. We suggest that the overexpression of IL-6 and TP53 may be a specific feature in rotator cuff disease patients with hypercholesterolemia.

Identification of the Interaction between Insulin-like Growth Factor Binding Protein-4 (IGFBP-4) and Heterogeneous Nuclear Ribonucleoprotein L (hnRNP L) (IGF결합 단백질-4(IGFBP-4)와 이질 핵 리보핵산단백질 L (hnRNP L)의 상호결합의 식별)

  • Choi, Mieyoung
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1311-1316
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    • 2013
  • Heterogeneous nuclear ribonucleoprotein L (hnRNP L) is a major pre-mRNA binding protein and it is an abundant nuclear protein that shuttles between the nucleus and the cytoplasm. hnRNP L is known to be related to many cellular processes, including chromatin modification, pre-mRNA splicing, mRNA export of intronless genes, internal ribosomal entry site (IRES)-mediated translation, mRNA stability, and spermatogenesis. In order to identify the cellular proteins interacting with hnRNP L, this study performed a yeast two-hybrid screening, using a human liver cDNA library. The study identified insulin-like growth factor binding protein-4 (IGFBP-4) as a novel interaction partner of hnRNP L in the human liver. It then discovered, for the first time, that hnRNP L interacts specifically with IGFBP-4 in a yeast two-hybrid system. The authenticity of this two-hybrid interaction of hnRNP L and IGFBP-4 was confirmed by an in vitro pull-down assay.

Effects of Astragalus Polysaccharides, Achyranthes bidentata Polysaccharides, and Acantbepanax senticosus Saponin on the Performance and Immunity in Weaned Pigs

  • Kang, P.;Xiao, H.L.;Hou, Y.Q.;Ding, B.Y.;Liu, Y.L.;Zhu, H.L.;Hu, Q.Z.;Hu, Y.;Yin, Y.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.6
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    • pp.750-756
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    • 2010
  • Two trials were conducted to study the effects of two Chinese herbal polysaccharides, Astragalus polysaccharides (APS) and Achyranthes bidentata polysaccharides (ABPS), and one Chinese herbal saponin, Acantbepanax senticosus saponin (ASS), on the immunity and growth performance of weaned pigs. Experiment 1 was a 14-day growth assay, in which 32 weaned pigs were randomly allocated to one of four dietary treatments: i) 0.05% talcum powder control; ii) 0.05% APS; iii) 0.05% mixture of APS and ASS in a 1:1 ratio by weight; and iv) 0.05% mixture of APS, ASS, and ABPS in a ratio of 1:1:1 by weight. Blood samples were collected on day 14 to determine plasma parameters. Feed intake, body weight gain, and feed efficiency were also determined. Experiment 2 was a 21-day immunity assay, in which 16 weaned pigs were randomly allotted to one of two dietary treatments: i) 0.05% talcum powder control; and ii) 0.05% mixture of APS and ASS in a 1:1 ratio by weight. On day 21, pigs were challenged with lipopolysaccharide (LPS) and 3 h later blood samples were collected and analyzed for lymphocyte proliferation as well as interleukin 6 (IL-6), insulin-like growth factor 1 (IGF-1), growth hormone (GH), and cortisol levels. In Experiment 1, feeding Chinese herbal polysaccharides and saponin increased growth performance of the pigs. The effects of the mixture of APS and ASS were especially notable, as there was a significant improvement in growth performance compared with the control (p<0.05). The plasma concentration of immunoglobulin G (IgG), nitric oxide (NO), and nitric oxide synthase (NOS) were increased in all treatments groups, with the mixture of APS and ASS increasing the level of IgG and NOS significantly (p<0.05), compared with the control. There was no difference in the NO level between the control and treatment groups (p>0.05). In Experiment 2, Chinese herbal polysaccharides and saponin showed immunostimulating effects. The level of cortisol, GH, and IGF-I were significantly increased (p>0.05), and the level of IL-6 showed a significant decrease (p<0.05) in the APS and ASS treatment after the LPS challenge. The mixture of APS and ASS could stimulate the blood lymphocyte proliferation significantly whether the LPS was injected or not (p<0.05). These results show that Chinese herbal extracts can improve growth performance and stimulate immunity of weaned pigs. A mixture of APS and ASS, compared with APS alone, could be a new kind of immunostimulant for weaned pigs, which could result in greater positive effects on their growth performance and immunity.

EFFECT OF ZIZYPHI FRUCTUS EXTRACT ON THE BIOLOGICAL ACTIVITY OF GINGIVAL FIBROBLAST (대조 추출물분획이 치은 섬유아세포의 생물학적 활성화에 미치는 영향)

  • Yang, Chang-Ho;Lee, Yong-Moo;Cho, Ki-Yeong;Bae, Ki-Hwan;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.24 no.1
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    • pp.144-154
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    • 1994
  • Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

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Intensive Proteomic Approach to Identify Secreted Peptides/Proteins from 3T3-L1 Adipocytes using Gel Electrophoresis and Liquid Chromatograph Separation Methods (젤 전기영동 및 액체 크로마토그래피 분리 방법을 이용하여 지방 세포로부터 분비되는 단백질들에 대한 프로테오믹스 연구 방법)

  • Hwang, Hyun-Ho;Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.55 no.3
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    • pp.203-212
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    • 2011
  • Adipocytes have been known to secrete a number of important proteins called adipokines with roles in energy metabolism, reproduction, cardiovascular function and immunity. In this study we have attempted to identify intensively secretory proteins from 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into mature adipocytes and then the cells were left in serum-free medium. The supernatant was filtrated and dialyzed. Lyophilized secretome was fractionated by two different methods, 1-D SDS PAGE and RP-FPLC. The tryptic peptides from the gel slices and the FPLC fractions were analyzed by nanoLC/ESI-MS/MS. We identified a total of 303 identical proteins from two methods, 251 proteins from 1-D gel and 184 proteins from RP-FPLC. 86 of them were listed as a secretory protein Finally, we identified many known or unknown secreted proteins existed in the low level including adiponectin, angiotensinogen, bone morphogenetic protein-1 (BMP-1), macrophage migration inhibitory factor (MIF), insulin like growth factor-II (IGF-II), interleukin-6 (IL-6), follistatin-related protein-1, minecan, and resistin. The existence of some of secreted proteins has been confirmed in RNA level. This proteomic experiment is useful for the intensive screening of secretory proteins in many kinds of other cells.

Effects of Boheo-tang and Boheo-tang plus Cervi Pantotrichum Cornu : on Lactation in Postpartum mice (보허탕(補虛湯)과 보허탕가녹용(補虛湯加鹿茸)이 산후 생쥐의 유즙분비에 미치는 영향)

  • Lee, Eun-Hee;Kim, Tae-Hee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.4
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    • pp.1-16
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    • 2008
  • Purpose: The purpose of this study is to investigate the effect of Boheo-tang (B) and Boheo-tang plus cervi pantotrichum cornu (B+CP) on lactation in postpartum C57BL/6N mice. Methods: Normal saline(control), Band B+CP (8mu l/g$) were administerd p.o. twice a day for 20 days. Lactating mammary gland tissues were examined through light microscope by the way of HE staining and immunohistochemical assay. Milk producing associated gene expression were accessed by RT-PCR. Results: In mammary gland, amount of adipose tissues were decreased in both Band B+CP treated group. And the ductal branches and alveolar tissues increased in both treated group. Immunoreactivity of prolactin receptors was increased both treated group, and immunoreactivity of oxytocin receptors was increased in the B+CP treated group. In both treated group, IGF-l mRNA expression was increased and TGF-$\beta$ mRNA expression was decreased. And PRL mRNA expression was increased in the B+CP treated group. PL-l mRNA expression was decreased in the B treated group but increased in the B+CP treated group. Conclusion: This study shows that treatment of Boheo-tang and Boheo-tang plus cervi pantotrichum cornu can improve postpartum lactation in C57BL/6N mice.

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Evaluation of the optimal dissolved oxygen level for rainbow trout (Oncorhynchus mykiss) in the recirculating aquaculture system (순환여과 양식시스템 내 무지개송어(Oncorhynchus mykiss)의 적정 용존산소 농도평가)

  • Kunhong PARK;Jinseo CHOI;Younghun LEE;Jeonghwan PARK
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.59 no.4
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    • pp.387-398
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    • 2023
  • Conventional aquaculture faces declining productivity, shifting to recirculating aquaculture system (RAS), known for minimizing water usage and maintaining consistent water temperatures for year-round fish growth. Rainbow trout (Oncorhynchus mykiss), a globally important cold-water species and the third most farmed fish in inland waters of Korea, valued for its fecundity and rapid growth. Dissolved oxygen, an important environmental factor affecting fish production and economics, highlights the need for smart aquaculture practices. Since 2018, the rise of intelligent aquaculture platforms, incorporating information and communications technology (ICT), emphasizes the essential role of RAS implementation. This eight-week study aimed to determine the optimal dissolved oxygen concentration for rainbow trout in RAS, utilizing a device for continuous monitoring, control and record. Dissolved oxygen concentrations were set at 5-6 mg/L, 9-10 mg/L, 14-15 mg/L and 17-18 mg/L. The growth rate significantly decreased at 5-6 mg/L, with no significant differences in other experimental groups. In hematological analysis, growth hormone (GH) was significantly highest at 5-6 mg/L, followed by 9-10 mg/L while Insulin-like growth factor-1 (IGF-1) was significantly lowest at 5-6 mg/L. In conclusion, the optimal dissolved oxygen concentration for rainbow trout in RAS is approximately 9-10 mg/L. Higher concentrations do not contribute to further growth or profitability.

Growth Associated Hormones Response and Fat Metabolism Change in Finishing Pigs Fed with n-Methyl-d, L-Aspartate

  • Xi, Gang;Xu, Zirong;Xiao, Ping
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1026-1030
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    • 2002
  • A trial was conducted to investigate the effect of dietary NMA on several growth associated hormones and fat metabolism in finishing pigs. A total of 84 crossbred finishing pigs (average initial BW of $56{\pm}$0.37kg) were divided into 6 pens, 14 pigs per pen (7 gilts and 7 barrows per pen). 3 pens of pigs were fed with control diet (corn-soybean meal) and the others were fed control diet addition with 50 mg/kg NMA. During the trial, all pigs were given free access to feed and water. After 44 days trial, 8 pigs from each treatment (4 gilts and 4 barrows, weight similar to average group weight, $86.94{\pm}0.71kg$ for control group, and $90.55{\pm}1.51kg$ for NMA treated group) were sacrificed to collect the sample of the liver, longissimus muscle, subcutaneous fat (10th rib). The addition of NMA in diet increased the IGF-I, Insulin, T3, T4 levels in serum by 50.68% (p<0.05), 38.36% (p<0.05), 123.33% (p<0.01), 60.58% (p<0.03), respectively. Meanwhile, IGF-I level in the liver and the muscle were increased with 17.83% (p<0.03) and 26.00% (p<0.03) with addition of NMA. The data from subcutaneous fat (10th rib) analysis showed that supplement of 50 mg/kg NMA decreased the total activities of malic dehydrogenase (MDH) by 20.54% (p<0.05), glucose-6- phosphate dehydrogenase (G-6-DPH) by 16.97% (p<0.05), and decreased the specific activities of MDH and G-6-DPH by 37.46% (p<0.01) and 35.06% (p<0.01), respectively. The hormone sensitive lipase (HSL) total activity was increased by 25.00% (p<0.05) in NMA treated pigs. These results indicated that addition of 50 mg/kg NMA to diet can induce the endocrine great change in finishing pigs, furthermore, inhibit the fat synthesis through suppressing lipogenic enzymes and promote the fat degradation by elevating HSL activity in finishing pigs.

Randomized, Double-blind, and Placebo-controlled Human Trial to Evaluate the Efficacy and Safety of Allium Fistulosum L. Root Extract on Improvement of Child Height Growth: Study Protocol (총백추출물의 어린이 키 성장에 대한 유효성 및 안전성을 평가하기 위한 무작위배정, 이중눈가림, 위약 대조 인체적용시험: 인체적용시험 프로토콜)

  • Shim, Soo Bo;Ko, Byoung Seob;Ryuk, Jin Ah;Lee, Jung Hwan;Lee, Ho Bong;Ha, Ki Chan;Kim, Yeung Mi;Lee, Hye Lim
    • The Journal of Pediatrics of Korean Medicine
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    • v.35 no.2
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    • pp.11-20
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    • 2021
  • Objectives The purpose of the study is to evaluate the safety of the Allium Fistulosum extract in children and its effectiveness in height growth. Methods This study is randomized, double-blind, placebo-controlled trial. The participants are children between the 3rd and 25th percentiles in height, and between the ages of 5 and 12 years. They are randomly assigned to treatment group or control group. The treatment group will take 5 g (1 g as Allium Fistulosum extract) for 24 weeks, 1 time a day. The control group will take the 5 g (0 g as Allium Fistulosum extract) of placebo for 24 weeks, 1 time a day. The primary outcome is change in height, and the secondary outcomes are growth rate, height standard deviations, Insulin-like growth factor-1 (IGF-1), Insulin-like growth factor binding protein-3 (IGFBP-3), IGF1-1/IGFBP-3 ratio, growth hormone, bone age, osteocalcin, and Z-score for growth. Results This protocol has been approved by the institutional review board (IRB) of Daejeon Korean Medicine Hospital of Daejeon University (IRB No. DJDSKH-20-BM-15), and registered in the Clinical Research Information Service (CRIS) (Registry No. KCT0005981). Conclusions This study will provide clinical information about the effectiveness and safety of Allium Fistulosum extract in children for their growth.