• Title/Summary/Keyword: ST2 protein

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Influence of the lung mechanical ventilation with injurious parameters on 7-ketocholesterol synthesis in Sus Scrofa

  • Klimenko, Oxana V.;Vobruba, Vaclav;Martasek, Pavel
    • BMB Reports
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    • v.43 no.4
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    • pp.257-262
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    • 2010
  • The aim of work was to investigate changes of 7-ketocholesterol synthesis in alveolar macrophages in the dynamic of lung mechanical ventilation with injurious parameters. The goal of in vitro part of work was to observe influence of 7-ketocholesterol on iNOS and MIP1 $\beta$ production in bronchoalveolar lavage fluid (BALF) cells. We used 17 healthy domestic pigs randomly assigned into two treatment groups: group I with mechanical ventilation with physiological parameters; group II underwent injurious ventilation with high volume tidal (VT) and low positive end expiratory pressure (PEEP). Cells were analyzed for CYP27A1 protein and gene expression levels, 7-ketocholesterol production. In alveolar macrophages of group II, we obtained increase of production of CYP27A1 protein and 7-ketocholesterol, as well as the expression of the CYP27A1 gene at the 2nd hour of ventilation. In the in vitro experiments we show dose-dependent increase of MIP1 $\beta$ and decrease of CYP27A1, iNOS protein production after 7-ketocholesterol treatment.

Effects of Selaginella Tamariscina on Apoptosis via the Activation of Caspase-3 in HL-60 (권백의 Caspase-3 활성화를 통한 HL-60 세포에서 세포사멸 유도효과)

  • Nam Hang Woo;Lee Sung Won;An Byung Sang;Chough Won Joon;Kim Yeong Mok;Mun Yean Ja;Ahn Seong Hun;Woo Won Hong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.751-758
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    • 2003
  • In our previous studies, we reported that Selaginella Tamariscina(ST) induced apoptotic cell death in HL-60 cells selectively. The cell viability after treatment with extract of ST was quantified by MTT assay and trypan bleu exclusion method. The results showed that application with ST in HL-60 induced 40% cell death at the concentration of 400 ㎍/ml. The cancericidic effect of Selaginella Tamariscina was mediated by apoptosis. Thus, HL-60 cells exposed to Selaginella Tamariscina displayed the DNA fragmentation ladder and nucleus chromatin condensation characteristic for apoptosis. The enzyme activity of caspase-3 and actived caspase-3 protein were markedly increased in HL-60 cells treated with the extract of Selaginella Tamariscina. In addition, the extract of Selaginella Tamariscina induced cleavage of PARP, a known substrate for caspase-3. The expression of Bcl-2, anti-apoptotic protein, was decreased by treatment of the aqueous extract of Selaginella Tamariscina in a dose-dependent manner. And the expression of pro-apoptotic Bax protein was increased. In conclusion, our results suggest that the extract of Selaginella Tamariscina may induce the apoptotic death of HL-60 cells via activation of caspase-3, cleavage of PARP protein, depletion of cellular ATP levels and Bcl-2 degradation.

Forage Productivity of Two Sorghum and a Sorghum-Sudangrass Hybrids harvested at Different Growth Stages (수확기에 따른 수수와 수수-수단그라스 교잡종의 사료생산성)

  • 이석순;최상집;김태주
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.11 no.2
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    • pp.121-128
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    • 1991
  • Forage productivity of two sorghum hybrids, Pioneer(P) 931 and P 956 and a sorghum-sudangrass hybrid, P 988, was studied. The 1st-cut of sorghun hybrids was made for silage 6 times from heading stage at a week-interval and regrowth (2nd-cut) was harvested on Oct. 13 although plants were not reached the same growth stage of 1st-cut. However, the 1st- and 2nd-cut of sorghum-sudangrass were made for green fodder 6 times from 4 weeks before heading stage at a week-interval. 1. Percent dry matter(DM) of 1st-cut of sorghum and 1st- and 2nd-cut of sorghum-sudangrass increased as growth stage advanced. Percent DM of 2nd-cut of sorghum decreased as the 1st-cut date delayed due to insufficient growing period on Oct. 13. 2. In the 1st-cut of sorghum the proportion of leaf blade (LB) or culm+leaf sheath (LS) to total DM decreased, but that of panicle increased as growth stage advanced. The panicle proportion of P 956 increased and culm+LS decreased more rapidly compared with those of P 931. IIowever, in sorghum-sudangrass hybrid the proportion of LB decreased and culmf LS increased as growth stage advanced through heading stage. 3. In the 1st-cut of sorghum hybrids crude(C) protein and C. fiber decreased, but nitrogen free extract (NFE) increased as growth stage advanced although C. fat and C. ash were similar among the growth stages. In the 2nd-cut of sorghum C. protein and C. ash increased, but NFE decreased as harvesting date delayed due to plants were younger although C. fat and C. fiber were similar among the harvesting dates. In the sorghum-sudangrass C. protein of 1st-cut decreased, but C. fiber of 1st-cut and NFE of the 1st-and 2nd-cut increased as growth stage advanced. 4. DM yield of 1st-cut of sorghum increased from heading stage to 3 weeks after heading and then levelled off, but total DM including regrowth was similar among the harvesting time. In the sorghum-sudangrass hybrid DM yield of 1st-cut and total DM yield including 2nd-and 3rd-cut increased as the harvestingtime delayed.

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Enhanced Sialylation of Recombinant Erythropoietin in CHO Cells by Human Glycosyltransferase Expression

  • Jeong, Yeon-Tae;Choi, One;Lim, Hye-Rim;Son, Young-Dok;Kim, Hong-Jin;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1945-1952
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    • 2008
  • Sialylation, the attachment of sialic acid residues to a protein, can affect the biological activity and in vivo circulatory half-life of glycoproteins. Human ${\alpha}2$,3-sialyltransferase (${\alpha}2$,3-ST) and ${\beta}1$,4-galactosyltransferase (${\beta}1$,4-GT) are responsible for terminal sialylation and galactosylation, respectively. Enhanced sialylation of human erythropoietin (EPO) by the expression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT was achieved using recombinant Chinese hamster ovary (CHO) cells (EC1). The sialic acid content and sialylation of N-glycans were evaluated by HPLC. When ${\alpha}2$,3-ST was expressed in CHO cells (EC1-ST2), the sialic acid content (moles of sialic acid/mole of EPO) increased from 6.7 to 7.5. In addition, the amount of trisialylated glycans increased from 17.3% to 26.1 %. When ${\alpha}2$,3-ST and ${\beta}1$,4-GT were coexpressed in CHO cells (EC1-GTST15), the degree of sialylation was greater than that in EC1-ST2 cells. In the case of EC1-GTST15 cells, the sialic acid content increased to 8.2 and the proportion of trisialylated glycans was markedly increased from 17.3% to 35.5%. Interestingly, the amount of asialoglycans decreased only in the case of GTST15 cells (21.4% to 14.2%). These results show that coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT is more effective than the expression of ${\alpha}2$,3-ST alone. Coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT did not affect CHO cell growth and metabolism or EPO production. Thus, coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT may be beneficial for producing therapeutic glycoproteins with enhanced sialylation in CHO cells.

Pharmacopuncture of Anti-inflammatory Herbal Compounds Suppresses Colon Inflammation-induced c-Fos like Protein Expression in Rats (소염(消炎) 약침(藥鍼)이 대장염 유발 흰쥐의 c-Fos 단백 발현에 미치는 효과)

  • Song, Jeong-Bang;Sohn, In-Chul;Ahn, Seong-Hun;Kim, Jae-Hyo
    • Journal of Pharmacopuncture
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    • v.13 no.3
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    • pp.47-62
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    • 2010
  • Objectives: Colitis is an inflammatory bowel disease characterized by colonic mucosal inflammation and chronic relapsing events represents. The purpose of this study is to evaluate effects of pharmacopuncture of anti-inflammatory herbal compound (AiC) applied to the different acupoints in the acute colitis induced by trinitrobenzenesulphonic acid (TNBS) intracolonic injection in rats. Methods: In Male Sprague - Dawley rats, weighing 250~400g, TNBS (5 mg/kg) was infused intrarectally through a silicon rubber catheter into the anus under isoflurane anaesthesia. Acupoints of LI4 (Hapkok), ST25 (Cheonchu), ST36 (Joksamni), and BL25 (Daejangsu) were intramuscularly injected by AiC, respectively (injection volume & times: 0.2 ml / acupoint, twice times on the 2nd & 3rd day). Expressions of cFos protein in the periaqueductal gray (PAG), locus coeruleus (LC), nucleus of solitary tract (Sol), and the 6th lumbar spinal cord (L6 s.c.) were observed at 24 hr after TNBS induced colitis by immunohistochemistry. Results: The expression of c-Fos protein in the L6 s.c., Sol, LC and PAG increased 24 hr after TNBS injection into colorectum as compared to normal and 50% ethanol treated group. AiC to LI4 inhibited the expression of c-Fos protein in Sol and PAG but not L6 s.c. and LC. AiC to ST36 showed significant inhibition the c-Fos expression in L6 s.c., Sol and PAG. AiC to ST25 only showed the effects in L6 s.c. and PAG. AiC to BL25 inhibited significantly the expression of c-Fos protein all over the areas. To investigate whether or not endogenous opioids are involved, intrathecal injection of naltrexone (30ug/30ul) was applied before the 2nd pharmacopuncture treatment 24 hr after TNBS-induced colitis in rat. Naltrexone reversed the inhibition of c-Fos protein expression in the spinal cord and brainstem. Conclusions: These data show that pharmacopuncture of Aic potently inhibits signal pathways ascending hypersensitivity of colorectum after TNBS induced colitis and depends on the endogenous opioids according to acupoints.

Zinc Deficiency Elevates Fecal Protein, But Not Electrolyte and Short-Chain Fatty Acid, Levels in Enterotoxigenic Escherichia coli-Induced Diarrhea in Rats

  • David, Ebuka E.;Yameen, Muhammad A.;Igwenyi, Ikechuku O.;David, Chidinma N.;Nwobodo, Valentine;Ismail, Akindele K.
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.25 no.1
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    • pp.79-86
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    • 2022
  • Purpose: To determine the effect of zinc deficiency on fecal protein, electrolyte, and short-chain fatty acid levels in both heat-stable (ST) and heat-labile (LT) enterotoxigenic Escherichia coli (ETEC)-induced diarrhea in rats. Methods: Albino rats, weighing 100 to 150 g, were divided into 2 groups, with 15 animals each: non-zinc and zinc-deficient. These two groups were sub-divided into three sub-groups with five rats each: control (saline); LT-ETEC; and ST-ETEC. Sodium phytate (30 mmol/L) was added to the animals' water to induce zinc deficiency, while diarrhea was induced using 5×109 ETEC cells/mL. Fecal protein levels were estimated using the Bradford method, while sodium and potassium levels were determined using atomic absorption spectrophotometry. Short-chain fatty acids were measured using gas chromatography-mass spectrometry. Results: Among the non-zinc and zinc-deficient groups, there were significant increases (p=0.04), (p=0.03) in fecal protein concentrations (mg/mL) in the LT-ETEC- (4.50±0.33), (6.50±0.26) and ST-ETEC- (3.85±0.19), (5.98±0.32) induced groups compared to the control groups (2.60±0.52), (3.50±0.11) respectively. Fecal sodium and potassium levels (mg/L) were significantly (p=0.029) increased in non-zinc-deficient rats induced with LT-ETEC (9.35±0.95, 1.05±0.48), and ST-ETEC (9.96±1.02, 1.21±0.45) compared with the control group (8.07±0.44, 0.47±0.17) but the increase were not statistically significant (p=0.059) in the zinc deficient rat groups. Fecal acetate and propionate levels (mg/g) significantly (p=0.032) increased when induced with LT-ETEC and ST-ETEC in non-zinc and zinc-deficient groups compared with the control groups. Conclusion: Zinc deficiency among rats with ETEC-induced diarrhea elevated fecal protein loss but may not have an effect on fecal sodium, potassium and short-chain fatty acid levels.

Effects of Curculiginis Rhizoma Pharmaco-Acupuncture at $ST_{36}$ on Monosodium Iodoacetate(MIA)-induced Osteoarthritic Rats (족삼리의 선모(仙茅)약침이 Monosodium Iodoacetate로 유발한 골관절염 흰쥐에 미치는 영향)

  • Ji, Min Jung;Lim, Seong Chul;Kim, Jae Soo;Lee, Hyun Jong;Lee, Yun Kyu
    • Journal of Acupuncture Research
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    • v.32 no.1
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    • pp.53-66
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    • 2015
  • Objectives : This study was designed to investigate effects of Curculiginis Rhizoma pharmaco-acupuncture at $ST_{36}$ on monosodium iodoacetate-induced osteoarthritic rats. Methods : Twenty rats were divided into four groups consisting of 5 rats: rats receiving no injection(normal), rats injected with monosodium Iodoacetate(MIA, control), rats injected with MIA and normal saline(N-S), and rats injected with MIA and Curculiginis Rhizoma (CRPA). N-S and CRPA were administered once a day at $ST_{36}$ during 21 days. After that we examined the weight-bearing ability of hind paws, liver and kidney function, immunocell, cytokines, proteins, and gene expression of cytokines. Injury of synovial tissue was measured by H & E, Safranin O immunofluorescence. Results : The weight-bearing ability of the hind paws, Serum TNF-${\alpha}$, IL-$1{\beta}$, IL-6, PGE2, LTB4, DPD, Osteocalcin, Protein COX-2 of CRPA decreased significantly. Protein Arachidonate 5 lipoxygenase of CRPA was decreased, but not significantly. Expression of gene COX-2, TNF-${\alpha}$, IL-$1{\beta}$, IL-6, NOS2 of CRPA decreased. In histological observations, CRPA was improved, compared with other control groups. Conclusions : It can be suggested that Curculiginis Rhizoma pharmaco-acupuncture at $ST_{36}$ has anti-inflammatory and pain relief effects on monosodium iodoacetate-induced osteoarthritic rats.

가미귀용탕(加味歸茸湯) 및 육미지황탕(六味地黃湯) 약침(藥鍼)이 생쥐의 성장(成長)에 미치는 영향(影響)

  • Kim, Min-Jung;Hong, Kwon-Eui
    • Journal of Pharmacopuncture
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    • v.9 no.3 s.21
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    • pp.105-116
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    • 2006
  • Objectives : To investigate the effect of Ga-mee-Gwui-ryong-tang herbal acupuncture(GR-HA) at GB39 and ST36 and the effect of Yuk-mee-ji-hwang-tang herbal acupuncture(YM-HA) at GB39 and KI10 on growth in mice. Methods : Mice were divided into 7 group(n=10) ; saline-ST36, saline-GB39 and KI10, GR-HA-ST36 group treated with GR-HA at right ST36, GR-HA-GB39 group treated with GR-HA at right GB39, YM-HA-GB39 group treated with YM-HA at right GB39, YM-HA-KI10 group treated with YM-HA at right KI10. Saline-ST36 group, saline-GB39 group and saline-KI10 group were injected with saline at right ST36, GB39, KI10 respectively. Saline injection and HA were performed 3 times a week for 18 days. The length(total height, body length and tail length), weight and the protein efficiency ratio were measured as well as IGF-1 level in serum. Results : 1. GR-HA at GB39 and GR-HA at ST36 promoted growth in weight, tail length and total height but there was no statistical significance. 2. GR-HA at GB39 significantly promoted protein efficiency on the 14th day and GR-HA at ST36 on the 3rd day. 3. GR-HA at GB39 significantly increased serum IGF-1. GR-HA at ST36 increased IGF-1 slightly but with no statistical significance. 4. YM-HA at GB39 and YM-HA at KI10 promoted growth in weight, height, tail length and total height but there was no statistical relevance. 5. YM-HA at GB39 and YM-HA at KI10 significantly promoted protein efficiency on the 14th day. 6. YM-HA at GB39 and YM-HA at KI10 significantly increased serum IGF-1. Serum IGF-1 level of YM-HA-KI10 group was significantly higher than that of YM-HA-GB39 group. 7. Serum IGF-1 level of GR-HA-GB39 group was significantly higher than that of YM-HA-GB39 group. Conclusions : GR-HA and YM-HA both may promote growth. This study suggests that herbal acupuncture at GB39 is more effective with Gwui-ryong-tang than Yuk-mee ji-hwang-tang to promote growth. It also suggests that Yuk-mee ji-hwang-tang herbal acupuncture is more effective at KI10 than at GB39.

Feedback Control of Cyclooxygenase-2 Expression by Prostaglandin E2 in Rheumatoid Synoviocytes

  • Min, So-Youn;Jung, Young Ok;Do, Ju-Ho;Kim, So-Yang;Kim, Jeong-Pyo;Cho, Chul-Soo;Kim, Wan-Uk
    • IMMUNE NETWORK
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    • v.3 no.3
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    • pp.201-210
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    • 2003
  • Objective: The role of prostaglandin $E_2$ (PGE2) in the etiopathogenesis of immune and inflammatory diseases has become the subject of recent debate. To determine the role of PGE2 in rheumatoid arthritis (RA), we tested the effect of exogenous PGE2 on the production of cyclooxygenase-2 (COX-2) by rheumatoid synoviocytes. Methods: Fibroblast-like synoviocytes (FLS) were prepared from the synovial tissues of RA patients, and cultured in the presence of PGE2. The COX-2 mRNA and protein expression levels were determined by RT-PCR and Western blot analysis, respectively. The PGE2 receptor subtypes in the FLS were analyzed by RT-PCR. Electrophoretic mobility shift assay (EMSA) was used to measure the NF-${\kappa}B$ binding activity for COX-2 transcription. The in vivoeffect of PGE2 on the development of arthritis was also tested in collagen induced arthritis (CIA) animals. Results: PGE2 ($10^{-11}$ to $10^{-5}M$) dose-dependently inhibited the expression of COX-2 mRNA and the COX-2 protein stimulated with IL-$1{\beta}$, but not COX-1 mRNA. NS-398, a selective COX-2 inhibitor, displayed an additive effect on PGE2-induced COX-2 downregulation. The FLS predominantly expressed the PGE2 receptor (EP) 2 and EP4, which mediated the COX-2 suppression by PGE2. Treatment with anti-IL-10 monoclonal antibodies partially reversed the PGE2-induced suppression of COX-2 mRNA, suggesting that IL-10 may be involved in modulating COX-2 by PGE2. Experiments using an inducer and an inhibitor of cyclic AMP (cAMP) suggest that cAMP is the major intracellular signal that mediates the regulatory effect of PGE2 on COX-2 expression. EMSA revealed that PGE2 inhibited the binding of NF-${\kappa}B$ in the COX-2 promoter via a cAMP dependent pathway. In addition, a subcutaneous injection of PGE2 twice daily for 2 weeks significantly reduced the incidence and severity of CIA as well as the production of IgG antibodies to type II collagen. Conclusion: Our data suggest that overproduced PGE2 in the RA joints may function as an autocrine regulator of its own synthesis by inhibiting COX-2 production and may, in part, play an anti-inflammatory role in the arthritic joints.

AN INVESTIGATION OF IMMUNIZATION AGAINST SOMATOSTATIN BY MEASURING ANTIBODY TITRES, SOMATOSTATIN AND SOMATOTROPIN PROFILES IN GILTS

  • Du, Z.L.;Hacker, R.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.2
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    • pp.211-218
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    • 1993
  • The effect of active immunization against porcine somatostatin (SRIF-14) on somatostation and somatotropin secretion profile in 18 gilts was investigated. Gilts were assigned to the following treatments: control (sham injection, n = 6); bovine serum albumin (BSA) (injection of BSA with bacterial protein adjuvant, n = 6); SRIF (injection of BSA-SRIF-14 conjugate with bacterial protein adjuvant n = 6). Serum SRIF and pST were assayed from the blood samples taken on day 7 after the last immunization injection. Anti-SRIF antibody titres were assayed in weekly samples two weeks after the initial immunization to one week after the last immunization. Results revealed that the immunization protocol used in the present investigation failed to produce antibodies capable of neutralizing endogenous somatostatin. In addition, the porcine somatotropin assay revealed no significant differences in baseline pST concentration, mean peak amplitude and number of peaks during a 24 h secretory period among SRIF, BSA and control treatment. There were also no differences in SRIF baseline concentration, peak amplitude, and number of peaks during a 24 h secretory period among any of the three treatments. Circulating concentrations of pST and pSRIF were highly correlated (r = -0.09). Furthermore, anti-SRIF antibody titre was not detected in the serum of the gilts actively immunized against SRIF. These data, collectively, suggest that the protocol employed in the present investigation for active immunization against SRIF is not an effective method for changing SRIF and pST secretion profiles of the gilt and thus to enhance performance.