• Title/Summary/Keyword: GAPDH-S

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Proteome analysis of sorghum leaf and root in response to heavy metal stress

  • Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Lee, Dong-Gi;Sarker, Kabita;Lee, Moon-Soon;Xin, Zhanguo;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.24-24
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    • 2017
  • Heavy metals at toxic levels have the capability to interact with several vital cellular biomolecules such as nuclear proteins and DNA, leading to oxidative stress in plants. The present study was performed to explore the metal tolerance mechanism in Sorghum seedling. Morpho-physiological and metal ions uptake changes were observed prominently in the seedlings when the plants were subjected to different concentrations of $CuSO_4$ and $CdCl_2$. The observed morphological changes revealed that the plants treated with Cu and Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cu and Cd was markedly increased by treatment with Cu and Cd, and the amount of interacting ions taken up by the shoots and roots was significantly and directly correlated with the applied level of Cu and Cd. Using the 2-DE method, a total of 24 and 21 differentially expressed protein spots from sorghum leaves and roots respectively, 33 protein spots from sorghum leaves under Cd stress were analyzed using MALDI-TOF/TOF MS. However, the over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. The results obtained from the present study may provide insights into the tolerance mechanism of seedling leaves and roots in Sorghum under heavy metal stress.

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Effect of adrenalectomy on gene expression of adrenoceptor subtypes in the hypothalamic paraventricular nucleus

  • Kam, Kyung-Yoon;Shin, Seung Yub;Han, Seong Kyu;Li, Long Hua;Chong, Wonee;Baek, Dae Hyun;Lee, So Yeong;Ryu, Pan Dong
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.207-215
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    • 2004
  • It is well known that the hypothalamic-pituitary-adrenocortical (HPA) axis is under the negative feedback control of adrenal corticosteroids. Previous studies have suggested that glucocorticoids can regulate neuroendocrine cells in the paraventricular nucleus (PVN) by modulating catecholaminergic transmission, a major excitatory modulator of the HPA axis at the hypothalamic level. But, the effects of corticosteroids on the expression of adrenoceptor subtypes are not fully understood. In this work, we examined mRNA levels of six adrenoceptor subtypes (${\alpha}_{1A}$, ${\alpha}_{1B}$, ${\alpha}_{2A}$, ${\alpha}_{2B}$, ${\beta}_1$ and ${\beta}_2$) in the PVN of normal and adrenalectomized (ADX) rats. Total RNA ($2.5{\mu}g$) was extracted from PVN micropunches of brain slices ($500{\mu}m$) and analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The levels of corticotropin-releasing hormone (CRH) mRNA were increased in the ADX rats relative to normal rats, indicating that the PVN had been liberated from the negative feedback of corticosteroids. Among the six adrenoceptor subtypes examined, mRNA levels for ${\alpha}_{1B}$- and ${\beta}_1$-adrenoceptors were increased, but the level for ${\beta}_2$-adrenoceptors was decreased in the ADX rats. The mRNA levels for the other three subtypes and for the general and neuronal specific housekeeping genes, glyceroaldehyde-3-phosphate dehydrogenase (GAPDH) and N-enolase, respectively, were not changed in the ADX rats. In conclusion, the results indicate that adrenal steroids selectively regulate the gene expression of adrenoceptor subtypes in the PVN.

Eotaxin mRNA Expression in Bronchial Mucosa of Patients with Asthma (천식 환자의 기관지 조직에서 Eotaxin mRNA 발현에 관한 연구)

  • In, Kwang-Ho;Cho, Jae-Yun;Kang, Sae-Yong;Lee, Sang-Youb;Shim, Jae-Jeong;Kang, Kyung-Ho;Yoo, Se-Hwa;Na, Young-Soon;Kim, Han-Gyum
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.697-704
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    • 1998
  • Background: Asthma is a chronic inflammatory disease of the airways characterized by a marked infiltration of eosinophils in the bronchial mucosa. Asthmatic bronchial mucosa produces many factors described as being chemotactic for inflammatory cells. IL-5, RANTES, and MCP-1 alpha are the chemotactic factors for eosinophils, but their roles are controversial. Recently eotaxin that is a potent eosinophil chemoattractant cytokine was detected in a guinea-pig model of allergic airway inflammation, and human eotaxin was cloned. Eotaxin is a specific chemoattractant for eosinophils, but its role in asthma is not confirmed. We examined the in vivo expression of eotaxin in bronchi of asthmatic patients. Methods : 11 asthmatics and 2 normal controls were enrolled. All subjects were underwent bronchoscopy with bronchial biopsies in 2nd or 3rd carina. RNA extraction from biopsy samples was done by acid-guanidium method. Semi-quantitaive RT-PCR was done for evaluation of eotaxin mRNA expression The extent of eosinophil infiltration was evaluated by counting the eosinophils in submucosa in HPF of microscope. Results : Eotaxin mRNA expressed in symptomatic, uncontrolled asthma. Steroid inhibited expression of eotaxin mRNA in asthma. Expression of eotaxin mRNA correlated with eosinophil infiltration in bronchial tissues. Conclusion: Expression of eotaxin mRNA increases in uncontrolled asthma and eotaxin is involved in the recruitment of eosinophils.

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A Study of the Anti-inflammatory Effects of Abies nephrolepis MAX. Extract in RAW 264.7 Cells (RAW 264.7 세포에서 분비나무(Abies nephrolepis MAX.) 추출물의 항염 효과에 대한 연구)

  • Min-Jeong Oh;Hyeon-Ji Yeom;Jin-Young Lee
    • Journal of Life Science
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    • v.34 no.3
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    • pp.160-169
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    • 2024
  • In this study, Abies nephrolepis MAX. was divided into A. nephrolepis MAX. stem (AS) extract and A. nephrolepis MAX. leaf (AL) extract. Their anti-inflammatory abilities and applicability as cosmetic materials were determined. Tests of the cell survival rate measured using RAW 264.7 cells and extracts of AS and AL showed 97.8% and 95.6% cell viability at a 500 ㎍/ml concentration. To determine anti-inflammatory activity, we examined the inhibitory effects on the production of LPS-induced NO in RAW 264.7 cells by Griess assay. The results showed that the AS and AL extracts presented a concentration-dependent inhibition of NO production. The protein expression inhibitory effects of AS and AL extracts were measured by western blot at 25, 50, and 100 ㎍/ml concentrations. β-actin was used as a positive control. The results of western blot of extracts from AS showed that the expression inhibition rate of the iNOS protein was decreased by 50.1% at the 100 ㎍/ml concentration. Additionally, the results of western blot of AL extracts showed that the expression inhibition rate of COX-2 and iNOS protein was decreased by 66% and 8.2% at the 100 ㎍/ml concentration. The mRNA inhibitory effect was measured by RT-PCR at 25, 50, and 100 ㎍/ml concentrations. GAPDH was used as a positive control. Consequently, the iNOS mRNA expression effect by RT-PCR of AS extract demonstrated by RT-PCR decreased by 27.9% at the 100 ㎍/ml concentration, and the iNOS and IL-6 mRNA expression effect of AL extract measured by RT-PCR decreased by 48.6% and 48.7% at the 100 ㎍/ml concentration.

Anti-inflammatory Activities Verification of Ambrosia trifida L. extract in RAW 264.7 Cells (RAW 264.7 세포에서의 단풍잎돼지풀 추출물의 항염증 활성 검증)

  • Yoo, Dan-Hee;Lee, Jin-Young
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.79-89
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    • 2020
  • This study was performed to evaluate the anti-inflammatory activities of 70% ethanol extract from Ambrosia trifida L. (AT). The electron donating ability and ABTS+ radical scavenging ability of extract from AT was shown to be 84.1% and 92.5% at 1,000 ㎍/ml concentration. The astringent effect of extract from AT was shown to be 94.7% at 1,000 ㎍/ml. The anti- inflammatory activities of extract of AT were investigated using RAW 264.7 cells induced by lipopolysaccharide (LPS). The cell toxicity effect of AT extract on RAW 264.7 performed MTT assay. As a result of the measured cell toxicity effect, 90% or more was shown with cell viability at a 500 ㎍/ml concentration. In nitric oxide synthesis inhibition effect, it was shown that extract from AT concentration dependent inhibited nitric oxide production. The protein expression inhibitory effect of AT extract was measured by western blot at 25, 50, and 100 ㎍/ml concentration and the β-actin used as a positive control. Consequently, the inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 protein expression inhibitory effect was decreased by 8.6%, 25.1% at 100 ㎍/ml concentration. The phosphorylation of extracellular signal-regulated kinase 1/2, p38, c-Jun NH2-terminal kinase and Iκ-Bα protein expression inhibitory effect was a decreased dependent concentration. The mRNA expression inhibitory effect was measured by reverse transcription - polymerase chain reaction at 25, 50, and 100 ㎍/ml concentration and the glyceraldehyde-3-phosphate dehydrogenase used as a positive control. Consequently, the iNOS, COX-2, interleukin (IL)-1β, IL-6 and tumor necrosis factor-α mRNA expression inhibition effect was a decreased dependent concentration in an LPS-activated macrophage. In conclusion, AT extract may have some effects on inflammatory factors as potential anti-inflammatory agents and natural substance for cosmetics.