• Title/Summary/Keyword: COX-1, COX-2

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Development of New Molecular Markers for the Identification of Male Sterile Cytoplasm in Peppers (Capsicum annuum L.)

  • Min, Woong-Ki;Kim, Byung-Dong;Kim, Sung-Gil;Lee, Sang-Hyeob
    • Horticultural Science & Technology
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    • v.29 no.1
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    • pp.53-60
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    • 2011
  • Cytoplasmic male sterility (CMS) induced by mutant mitochondria genome, has been used for commercial seed production of $F_1$ hybrid cultivars in diverse crops. In pepper (Capsicum annuum L.), two sterile cytoplasm specific gene organization, atp6-2 and coxII were identified. An open reading frame, orf456 nearby coxII gene has been speculated to induce male sterility (MS) by mutagenic analysis. Moreover, molecular markers for atp6-2 and coxII of mitochondrial genotype (mitotype) were developed. However, the Cytoplasmic MS specific markers, atp6SCAR and coxIISCAR markers appeared in both N and S cytoplasms when polymerase chain reaction (PCR) cycles prolonged more than 40 cycles. Since the reported molecular markers were dominant markers, the presence of the faint sterile-specific band in normal cytoplasm may lead to the mis-classification of pepper breeding lines. To solve this problem, one common forward primer and two different reverse primers specific to normal coxII and sterile orf456 genes were designed after analyzing their gene organizations. By using these three primers, N and S coxII specific bands were co-amplified in male-sterile lines, but only normal coxII specific band was amplified in maintainer lines. Since the reverse primer for sterile coxII was specifically designed 275 bp downstream of orf456, relatively stable PCR amplification patterns were observed regardless of the number of PCR cycles. These primer sets easily identified different mitotypes among the divergent breeding lines, commercial cultivars and diverse germplasms.

Effect of bee venom on cell proliferation and cyclooxygenase 2 expression in the dentate gyrus of mice with acetic acid induced hyperalgesia (세포 증식과 COX 2 발현에 미치는 봉독의 효과)

  • Lyoo, Eun-kyoung;Choi, Do-young;Lee, Jae-dong
    • Journal of Acupuncture Research
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    • v.20 no.2
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    • pp.112-122
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    • 2003
  • 연구 배경 및 목적 : 봉독은 항염증 등의 효과를 가지고 있다고 알려져 있지만 세포 증식과 관련된 봉독의 효과에 대한 고찰을 위해 본 연구(硏究)에서는, 봉독(蜂毒) 약침(藥鍼) 자극(刺戟)이 아세트산 유발(誘發) 통각(痛覺) 과민증(過敏症)을 가진 쥐의 치상회(齒狀回)에서의 세포(細胞) 증식(增植)과 COX2 발현(發現)에 미치는 영향(影響)에 대해 알아보고자 하였다. 실험방법 : 대조군, 아세트산 처치군, 아세트산 0.1mg/kg 봉독 처치군, 아세트산 1mg/kg 봉독처치군(n=5 in each group)의 네 군으로 나누고 해당 군에 일벌 봉독(蜂毒)(Sigma Chemical Co., St. Louis, MO, USA)을 양측 족삼리 경혈(ST36)에 주입시키고 30분 후 아세트산(100% acetic acid 1% 용액의 0.5ml)을 복강내로 주입하여 복부 긴장 회수를 세었으며, BrdU 양성, COX 2 양성 세포수를 면역 화학 조직법을 수행하여 세어 보았다. 결과 : 아세트산 처치군에서는 대조군에 비해 5 bromo 2' deoxyuridine 양성 세포의 수는 감소(減少)되며, 치상회(齒狀回)에서의 COX 2의 발현(發現)은 증강(增强)되는 것으로 보여졌다. 봉독(蜂毒) 주입(注入)은 아세트산 유발(誘發) 복부(腹部) 긴장(緊張) 횟수와 치상회(齒狀回)에서의 COX2 발현(發現)을 억제(抑制)하여, 치상회(齒狀回)에서의 세포(細胞) 증식(增植)을 증가(增加)시켰다. 결론 : 이번 결과(結果)에서 보면, 치상회(齒狀回)에서의 COX 2의 발현(發現)은 세포(細胞) 증식(增植) 억제(抑制)와 관련(關聯)되며 봉독(蜂毒)은 COX 2 발현(發現) 억제(抑制)를 통해 치상회(齒狀回)에서의 새로운 세포(細胞) 형성(形成)을 증가(增加)시킨다는 것을 알 수 있다.

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Cyclooxygenase-2 over-expression is associated with increased mast cells in CCl4-induced hepatic fibrosis

  • Jekal, Seung-Joo;Lee, Jae-Hyoung;Park, Seung-Teack
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.4
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    • pp.229-238
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    • 2012
  • Cyclooxygenase(COX-2) is an inducible enzyme that catalyzes the synthesis of prostaglandins (PGs) from arachidonic acid. Over-expression of COX-2 has been reported to be associated with progressive hepatic fibrosis in chronic hepatic C infection and rat liver fibrosis induced by carbon tetrachloride($CCl_4$). Recently, it is well known that mast cell products can stimulate the proliferation of hepatic stellate cells and key players in liver fibrosis. But little is known regarding their role in $CCl_4$-induced liver fibrosis in rat. Our aim was to investigate the relation between COX-2 expression and mast cells during liver fibrosis after $CCl_4$ treatment. Thirty Wistar rats were divided into five groups (non-treated 0, 2, 4, 6 and 8-week after $CCl_4$-treatment). Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to assess the expression of ${\alpha}$-smooth muscle actin (${\alpha}$-SMA), collagen-1 and COX-2 in liver tissue from $CCl_4$-treated rats. The density of collagen and mast cells were determined using a computerized image analysis system in liver sections stained with picrosirius red and toluidine blue, respectively. The expression levels of ${\alpha}$-SMA, collagen-1 and COX-2 mRNA were significantly higher at 2 wk in $CCl_4$-treated groups than non-treated group. The number of mast cells in liver tissues increased gradually from 2 wk to 6 wk depending on the fibrosis severity but decreased abruptly at 8 wk. The significant increase of collagen-1 and ${\alpha}$-SMA mRNA expression in $CCl_4$-treated rats was continued until 6 wk while the COX-2 mRNA was significantly decreased at 8 wk. These results suggest that increased mast cells are closely associated with COX-2 over-expression during hepatic fibrogenesis of $CCl_4$-treated rats.

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Clinical Implication of Cyclooxygenase-2 Expression for Rectal Cancer Patients with Lymph Node Involvement (림프절 전이를 동반한 직장암 환자들에서 Cyclooxygenase-2 발현의 임상적 의미)

  • Lee, Hyung-Sik;Choi, Young-Min;Hur, Won-Joo;Kim, Su-Jin;Kim, Dae-Cheol;Roh, Mee-Sook;Hong, Young-Seoub;Park, Ki-Jae
    • Radiation Oncology Journal
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    • v.27 no.4
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    • pp.210-217
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    • 2009
  • Purpose: To assess the influence of cyclooxygenase-2 (COX-2) expression on the survival of patients with a combination of rectal cancer and lymph node metastasis. Materials and Methods: The study included rectal cancer patients treated by radical surgery and postoperative radiotherapy at the Dong-A university hospital from 1998 to 2004. A retrospective analysis was performed on a subset of patients that also had lymph node metastasis. After excluding eight of 86 patients, due to missing tissue samples in three, malignant melanoma in one, treatment of gastric cancer around one year before diagnosis in one, detection of lung cancer after one year of diagnosis in one, liver metastasis in one, and refusal of radiotherapy after 720 cGy in one, 78 patients were analyzed. The immunohistochemistry for COX-2 was conducted with an autostainer (BenchMark; Ventana, Tucson, AZ, USA). An image analyzer (TissueMine; Bioimagene, Cupertino, CA, USA) was used for analysis after scanning (ScanScope; Aperio, Vista, CA, USA). A survival analysis was performed using the Kaplan Meier method and significance was evaluated using the log rank test. Results: COX-2 was stained positively in 62 patients (79.5%) and negatively in 16 (20.5%). A total of 6 (7.7%), 15 (19.2%), and 41 (52.6%) patients were of grades 1, 2, and 3, respectively for COX-2 expression. No correlation was found between being positive of COX-2 patient characteristics, which include age (<60-year old vs. $\geq$60), sex, operation methods (abdominoperineal resection vs. lower anterior resection), degrees of differentiation, tumor size (<5 cm vs. $\geq$5 cm), T stages, N stages, and stages (IIIa, IIIb, IIIc). The 5-year overall and 5-year disease free survival rates for the entire patient population were 57.0% and 51.6%, respectively. The 5-year overall survival rates for the COX-2 positive and negative patients were 53.0% and 72.9%, respectively (p=0.146). Further, the 5-year disease free survival rates for the COX-2 positive and negative patients were 46.3% and 72.7%, respectively (p=0.118). The 5-year overall survival rates were significantly different (p<0.05) for the degree of differentiation, N stage, and stage, whereas the 5-year disease free survival rates were significant for N stage and stage. Conclusion: Being positive for and the degree of COX-2 expression did not have a significant influence on the survival of rectal cancer patients with lymph node metastasis. However, N stage and stage did significantly influence the rateof survival. Further analysis of a greater sample size is necessary for the verification of the effect of COX-2 expression on the survival of rectal cancer patients with lymph node involvement.

Inhibitory Effect of Benzofuran Compound on Cyclooxygenase

  • Min, Kyung-Rak;Ahn, Ki-Young;Chung, Eun-Yong;Lee, Yong-Rok;Kim, Yeong-Shik;Kim, Young-Soo
    • Natural Product Sciences
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    • v.10 no.6
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    • pp.315-320
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    • 2004
  • Alpha-viniferin was previously isolated as a cyclooxygenase (COX)-2 inhibitor from Carex humilis (Cyperaceae) and is an oligomeric stilbene compound with benzofuran (BF) moieties in its chemical structure. In the present study, a chemically synthetic BF compound, named as 3,3-dimethyl-2,3,4,6,7,8,9,10,11,12,13,14,15,16,17,18-hexadecahydro-1H-benzo[b] cyclopentadeca[d]furan-1-one, was discovered to inhibit bacterial lipo polysaccharide (LPS)-induced prostaglandin $E_2$ $(PGE_2)$ production in macrophages RAW 264.7. The BF compound exhibited a selectively preferred inhibitory effect on COX-2 activity over COX-1 activity. Furthermore, BF compound inhibited LPS-induced COX-2 expression at transcription level. As a down-regulatory mechanism of COX-2 expression shown by BF compound, suppression of nuclear factor $(NF)-{\kappa}B$ activation has been demonstrated. BF compound inhibited LPS-induced $NF-{\kappa}B$ transcriptional activity and nuclear translocation of $NF-{\kappa}B$ p65, in parallel, but did not affect LPS-induced degradation of inhibitory ${\kappa}B{\alpha}$ protein $(I{\kappa}B{\alpha})$. Taken together, anti-inflammatory effect of BF compound on $PGE_2$ production was ascribed by its down-regulatory action on LPS-induced COX-2 synthesis in addition to inhibitory action on enzyme activity of COX-2.

Inhibitory effect of Chelidonii Herba water extract on production of Nitric Oxide, Expression of iNOS and COX-2 in lipopolysaccharide-activated Raw 264.7 cells (백굴채(白屈菜)의 물추출물이 lipopolysaccharide로 유도된 Nitric Oxide의 생성 및 iNOS와 COX-2의 발현에 미치는 영향)

  • Zhao Rong-Jie;Kim Young-Woo;Byun Sung-Hui;Kim Sang-Chan
    • Herbal Formula Science
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    • v.12 no.2
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    • pp.163-173
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    • 2004
  • Chelidonii Herba (CHE, Baek-gul-chae in Korean), which has its original description in Gu-Hwang-Bon-Cho, a classic book of oriental Herbal book, is widely used in the treatment of stomach cancer, jaundice, gasrtic ulcer, edema and stomach pain, in Korea, Japan and China. The present study was conducted to evaluate the effect of CHE on the nitric oxide (NO) production, iNOS and COX-2 expression in lipopolysaccharide - activated Raw 264.7 cells. After the treatment of CHE, NO production was monitored by measuring the nitrite content in culture medium, cell viability was measured by MIT assay. COX-2 and iNOS were determined by lmmunoblot analysis. The production of nitric oxide was significantly inhibited by pretreatment (1h) with CHE (0.1-0.3 mg/ml) on LPS-activated Raw264.7 cells. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein were up-regulated by LPS, but the increased levels of iNOS and COX-2 were inhibited by pretreatment of CHE (0.1-0.3 mg/ml), respectively. Thus, the present data suggest that CHE may play an important role in adjunctive therapy in Gram-negative bacterial infections.

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Intraspecific variation of gene structure in the mitochondrial large subunit ribosomal RNA and cytochrome c oxidase subunit 1 of Pyropia yezoensis (Bangiales, Rhodophyta)

  • Hwang, Il Ki;Kim, Seung-Oh;Hwang, Mi Sook;Park, Eun-Jeong;Ha, Dong-Soo;Lee, Sang-Rae
    • ALGAE
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    • v.33 no.1
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    • pp.49-54
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    • 2018
  • Red algal mitochondrial genomes (mtDNAs) can provide useful information on species identification. mtDNAs of Pyropia / Porphyra (Bangiales, Rhodophyta) have shown diverse variation in their size and gene structure. In particular, the introns and intronic open reading frames found in the ribosomal RNA large subunit gene (rnl) and cytochrome c oxidase subunit 1 gene (cox1) significantly vary the mitochondrial genome size in Pyropia / Porphyra species. In this study, we examined the exon / intron structure of rnl and cox1 genes of Pyropia yezoensis at the intraspecific level. The combined data of rnl and cox1 genes exhibited 12 genotypes for 40 P. yezoensis strains, based on the existence of introns. These genotypes were more effective to identify P. yezoensis strains in comparison to the traditional DNA barcode cox1 marker (5 haplotypes). Therefore, the variation in gene structure of rnl and cox1 can be a novel molecular marker to discriminate the strains of Pyropia species.

Involvement of MAPK activation in chemokine or COX-2 productions by Toxoplasma gondii

  • Kim Ji-Young;Ahn Myoung-Hee;Song Hyun-Ouk;Choi Jong-Hak;Ryu Jae-Sook;Min Duk-Young;Cho Myung-Hwan
    • Parasites, Hosts and Diseases
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    • v.44 no.3
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    • pp.197-207
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    • 2006
  • This experiment focused on MAPK activation in host cell invasion and replication of T. gondii, as well as the expression of CC chemokines, MCP-1 and $MIP-1\alpha$, and enzyme, COX-2/prostaglandin $E_2(PGE_2)$ in infected cells via western blot, $[^3H]-uracil$ incorporation assay, ELISA and RT-PCR. The phosphorylation of ERK1/2 and p38 in infected HeLa cells was detected at 1 hr and/or 6 hr postinfection (PI). Tachyzoite proliferation was reduced by p38 or JNK MAPK inhibitors. MCP-1 secretion was enhanced in infected peritoneal macrophages at 6 hr PI. $MIP-1\alpha$ mRNA was increased in macrophages at 18 hr PI. MCP-1 and $MIP-1\alpha$ were reduced after treatment with inhibitors of ERK1/2 and JNK MAPKs. COX-2 mRNA gradually increased in infected RAW 264.7 cells and the secretion of COX-2 peaked at 6 hr PI. The inhibitor of JNK suppressed COX-2 expression. $PGE_2$ from infected RAW 264.7 cells was increased and synthesis was suppressed by PD98059, SB203580, and SP600125. In this study, the activation of p38, JNK and/or ERK1/2 MAPKs occurred during the invasion and proliferation of T. gondii tachyzoites in HeLa cells. Also, increased secretion and expression of MCP-1, $MIP-1\alpha$, COX-2 and $PGE_2$ were detected in infected macrophages, and appeared to occur via MAPK signaling pathways.

Suppression of Cyclooxygenase-2 Expression of Skin Fibroblasts by Wogonin, a Plant Flavone from Scutellaria Radix

  • Chi, Yeon-Sook;Kim, Hyun-Pyo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.96-96
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    • 2003
  • Previously, wogonin (5,7-dihydroxy-8-methoxyflavone) was found to suppress proinflammatory enzyme expression including cyclooxygenase-2 (COX-2), contributing to in vivo anti-inflammatory activity against skin inflammation. However, the detailed effect on each skin cell type has not been understood. Therefore, present investigation was carried out to find the effect of wogonin on inflammation-associated gene expression from skin fibroblasts in culture using reverse transcriptase-polymerase chain reaction. As a result, it was found that wogonin (10 - 100 ${\mu}$M) clearly down-regulated COX -2 expression from NIH/3T3 cells treated with 12-O-tetradecanoylphorbol 13-acetate, interleukin-1${\beta}$ or tumor necrosis factor-a. But, the expression levels of COX-1, interleukin-1${\beta}$ and fibronectin were not significantly affected. This finding was well correlated with significant reduction of prostaglandin E$_2$(PGE$_2$) production by wogonin. As a comparison, NS-398 (selective cyclooxygenase-2 inhibitor) did not suppress COX -2 expression and other gene levels, while PGE$_2$production was potently reduced at 0.1 - 10 ${\mu}$M. All these results suggest that COX -2 down-regulation of skin fibroblasts may be, at least in part, one of anti-inflammatory mechanisms of wogonin against skin inflammation.

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Cyclooxygenase-2 Expression in Urinary Bladder Transitional Cell Carcinoma and its Association with Clinicopathological Characteristics

  • Tabriz, Hedieh Moradi;Olfati, Golrokh;Ahmadi, Seyed Ali;Yusefnia, Sudabeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4539-4543
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    • 2013
  • Background: Transitional cell carcinoma (TCC) is the most predominant type of urinary bladder tumor. As cyclooxygenase (COX)-2 is recently introduced as an attractive target molecule in bladder TCC, we evaluated the immunohistochemical expression of this marker and its association with several clinicopathological characteristics. Materials and Methods: This cross-sectional study was performed in the Pathology department of Sina Hospital in Tehran, Iran during 2006-2011. Ninety-two paraffin embedded blocks were selected from patients with urinary bladder TCC who underwent cystectomy or transurethral resection (TUR). Then, we assessed COX-2 expression by immunohistochemical staining using antibody against COX-2. Staining in more than 5% of tumor cells was considered as positive expression. Results: COX-2 was expressed in 50 % of our patients. This marker was markedly expressed in high grade bladder TCC (62.1%) versus other grades and there was statistically a significant difference in COX-2 expression between various grades (p=0.008). In addition, patients' age, lymphatic and perineurial invasion were associated with the expression of COX-2 (p=0.001, 0.015 and 0.039, respectively). However, other parameters such as stage, tumor size, venous invasion and lymph node metastasis did not show any significant relationship with this marker (all, p>0.05). Conclusions: COX-2 was expressed in urinary bladder TCC especially in high grade forms, advocating its probable role in the differentiation of this tumor. Accordingly, COX-2 could be a valuable biological target molecule in the evaluation and treatment of patients with bladder TCC.