• Natural Product Sciences
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• 제3권1호
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• pp.19-28
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• 1997

Constitutive cyclooxygenase (COX-I) is present in cells under physiological conditions, whereas inducible cyclooxygenase (COX-II) is induced by some cytokines, mitogens, and endotoxin presumably in pathological conditions such as inflammation. We have evaluated the inhibitory effects of solvent fractionated extracts of natural products on the activities of COX-I and COX-II. Oxygen uptake COX assay was performed, as a primary screening from the tissue extracts of bovine seminal vesicles (BSV), by monitoring the initial rate of oxygen uptake using an oxygen electrode. Additionally, we evaluated plant extracts for the inhibitory effects of COX-I (in HEL cells) and COX-II (in lipopolysaccharide activated J774A.1 macrophages) using thin layer chromatography of prostanoids produced from $^{14}C-labelled$ arachidonic acid (AA). The use of such models of COX-I and COX-II assay will lead to the identification of specific inhibitors of cyclooxygenases with presumably less side effects than present therapies. Inhibitory effects of 50 kinds of plant extracts on the COX-I and COX-II activities were determined and the active fractions were found in the ethyl acetate fractions of Dryopteris crassirhizoma (roots), Amomum cardamomum (roots), Triticum aestivum (seeds), Perilla sikokiana (leaves), Anemarrhena asphodeloides (roots). Especially, the ethyl acetate fraction of Dryopteris crassirhizoma (roots), which exhibited the strong inhibition against BSV COX $(IC_{50},\;65.4\;{\mu}g/ml)$, COX-I $(IC_{50},\;8.5\;{\mu}g/ml)$, and COX-II $(IC_{50},\;17.2\;{\mu}g/ml)$, is under investigation to isolate active principles using activity-guided fractionation method.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4453-4458
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• 2012

Angiogenesis plays a significant role in colorectal cancer (CRC) and cyclooxygenase-2 (COX-2) appears to be involved with multiple aspects of CRC angiogenesis. Our aim was to investigate the inhibitory effects of Tan II-A (Tanshinone II-A, Tan II-A) on tumor growth in mice, as well as alteration of expression of COX-2 and VEGF in CRC. We established the mice xenograft model of C26 CRC cell line, and injected 0.5, 1, 2mg/kg of Tan II-A and 1mg/kg of 5-FU in respectively in vivo. Then, we assayed tumor weight and volume, and evaluated microvascular density and expression of VEGF. COX-2 promoter and COX-2 plasmids were transfected into HCT-116 cells, followed by detection of COX-2 promoter activity by chemiluminescence, and detection of COX-2 mRNA expression by fluorescence quantitative PCR. Taken together, the results showed Tan II-A could inhibit tumor growth and suppress the VEGF level in vivo. HCT-116 cell experiments showed marked inhibitory effects of Tan II-A on COX-2 and VEGF in a dose-dependent manner. The results indicate that Tan II-A can effectively inhibit tumor growth and angiogenesis of human colorectal cancer via inhibiting the expression level of COX-2 and VEGF.

• 원예과학기술지
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• 제29권1호
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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.

• Journal of Chest Surgery
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• 제32권3호
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• pp.249-254
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• 1999

배경: Cox-Maze 술식후의 좌심방의 수축력은 우심방에 비하여 회복률이 낮은 것으로 알려져 있다. 술전 좌심방의 크기가 큰 환자군에서의 Cox-Maze 술식은, 좌심방 후벽의 많은 부위를 전기전도계와 분리함으로써 술후 좌심방 기능의 회복을 저하시키는 한 요인이 될 수 있다. 본 연구자들은 좌심방이 심하게 확장된 경우에 좌, 우 폐정맥을 각각 분리하는 변형된 Cox-Maze 술식을 시행함으로써 이러한 술식이 술후 좌심방 기능의 회복에 도움을 주는지 여부에 대해 알아보고자 하였다. 대상 및 방법: 1995년 2월부터 1997년 10월까지 승모판막 질환과 동반된 심방세동으로 Cox-Maze 술식을 시행한 환자들 중, 수술후 6개월 이상의 추적관찰이 이루어지고 동율동 전환이 이루어진 환자를 대상으로 하였다. I군은 좌, 우 폐정맥사이의 거리가 6.5cm미만인 경우(n=30), II군은 좌, 우 폐정맥사이의 거리가 6.5cm 이상인 경우(n=16)로서 I, II군은 좌,우 폐정맥을 함께 분리하는 기존의 Cox-Maze 술식을 시행하였고, III군은 폐정맥사이의 거리가 6.5cm이상인 경우에 좌, 우 폐정맥을 각각 분리하는 변형된 Cox-Maze 술식을 사용하였다(n=9). 결과:각 군사이에는 좌심방 크기를 제외한 검사소견은 차이가 없었으며 III군의 경우 수술시 대동맥차단 시간이 유의하게 증가되었다(p<0.05). 술후 장기 추적 검사상 우심방의 수축력 회복은 I군에서 90%, II군에서 81%, 그리고 III군에서 100%에서 회복됨이 관찰되었다. 좌심방의 수축력회복은 I군에서 63%, II군에서 31%에서 회복되어 좌심방이 확장된 II군에서 유의한 감소가 있었으며 (p<0.04), III군에서는 66%에서 회복되었으나 I, II군과 유의한 차이는 없었다. 결론:이러한 양폐정맥 사이를 분리, 절개하는 변형된 Cox-Maze 술식은 좌심방 수축력의 회복에 긍정적인 영향을 미칠 것으로 추정되며, 향후 더 많은 환자군에서 연구가 시행되어야 하리라 사료된다.

• 대한의생명과학회지
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• 제15권2호
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• pp.113-118
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• 2009

Actin cytoskeletal architecture is believed to be a crucially important modulator of chondrocyte phenotype. 2DG(2-Dexoy-D-glucose) induces reorganization of actin cytoskeletal architecture in chondrocytes. In this study, we have investigated the effects of 2DG on dedifferentiation and inflammation via reorganization of cytoskeletal architecture in rabbit articular chondrocytes, with a focus on p38 kinase pathway. Treatment of 2DG alone reduced type II collagen and COX-2 expression in chondrocytes. But, 2DG reduced type II collagen was recovered by CD, disruptor of actin cytoskeletal architecture, whereas did not affect on COX-2 expression and production of $PGE_2$ compared with 2DG alone treated cells. Treatment of 2DG with JAS, inducer of cytoskeletal architecture polymerization, accelerated reduction of type II collagen expression and synthesis of proteoglycan but did not affect on COX-2 expression and production of $PGE_2$. Also, 2DG stimulated activation of p38 kinase. This result showed that 2DG regulates type II collagen but not cyclooxygenase-2 expression through reorganization of cytoskeletal architecture via p38 kinase pathway in rabbit articular chondrocytes.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.321.2-321.2
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• 2002

Possible role of anti-inflammatory effects of a new iridoids, patridoid I. II and II-A which were isolated from Patrinia saniculaefolia. examined by assessing their effects on tumor necrosis factor $\alpha$ (TN F$\alpha$) and 2 enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the lipopolysaccaride (LPS)-stimulated murine macrophage-like cell line RAW 264.7. Among them. patridoid II consistently inhibited the production of TNF$\alpha$ and NO production in a dose dependent manner. But patridoid I and patrioid ll isomer palrioid ll-A. these compounds very weakly inhibited NO producion. Moreover. treatment of macrophage with these compounds, the decrease in NO products was accompanied by a decrease in iNOS protein level as assessed by Western Blot. But these compounds did not affect COX-2 protein expression in LPS-stimulated macrophage. Our results suggest that patridoid ll could become a leading compound for developing a novel of anti-inflammalory drugs.

• Journal of Acupuncture Research
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• 제19권4호
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• pp.74-88
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• 2002

Objectives : This study is aimed to investigate the effects of bee venom and melittin on cell death in synovial cell line. Methods : It was evaluated by using MTT assay, morphologic method, DNA fragmenation, NO generation, flow cytometry, immunocytochemistry analysis, RT-PCR, Western blot. Results : The obtained results are summarized as follows: 1. The MTT assay demonstrated that synovial cell viability was significantly inhibitted dose-dependently by treatment with bee venom and melittin in comparison with control. 2. The morphologic study demonstrated that synovial cell showed apoptosis after treatment with bee venom and melittin for 6 hours using microscope. 3. In case of NO generation bee venom group and melittin group showed significant inhibition in comparison with control. 4. The Flow cytometry demonstrated that apoptosis of synovial cell treated with bee venom and melittin was related with stop of cell cycle in stage of $G_0/G_1$. 5. DNA fragmenation demonstrated that synovial cell treated with bee venom and melittin showed DNA ladder below l Kbp. 6. Immunocytochemistry assay demonstrated that COX-II and PLA2 were strongly down-regulated by treatment with bee venom and melittin whereas iNOS was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment. 7. RT-PCR analysis demonstrated that iNOS were strongly down-regulated by treatment with bee venom and melittin whereas COX-II was almostly not expressed by bee venom treatment and slightly expressed by melittin treatment. 8. Western blot demonstrated that iNOS were strongly down-regulated by treatment with $15{\mu}g/ml$ bee venom whereas COX-II was strongly down-regulated from $5{\mu}g/ml$ bee venom. Conclusions : These results suggest that bee venom and melittin have significant effect on cell death in synovial cell line and further study is needed in vivo.

• Molecules and Cells
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• 제20권3호
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• pp.416-422
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• 2005

We previously used Southern blot analysis to detect restriction-length polymorphisms between male fertile and cytoplasmic male sterile (CMS) cytoplasms at the coxII and atp6 loci of the mtDNA of Capsicum annuum L. Two copies of atp6 were found in each male fertile and CMS pepper lines. Interestingly, one of the copies of atp6 in CMS pepper was a 3'-truncated pseudogene. The open reading frame of the coxII gene was the same in the fertile (N-) and CMS (S-) lines. However, the nucleotide sequence in the S-cytoplasm diverged from that in the N-cytoplasm 41 bp downstream of the stop codon. To develop CMS-specific sequence-characterized amplified region (SCAR) markers, inverse PCR was performed to characterize the nucleotide sequences of the 5' and 3' flanking regions of mitochondrial atp6 and coxII from the cytoplasms of male fertile (N-) and CMS (S-) pepper plants. Based on these data, two CMS-specific SCAR markers, 607 and 708 bp long, were developed to distinguish N-cytoplasm from S-cytoplasm by PCR. The CMS-specific PCR bands were verified for 20 cultivars containing either N- or S-cytoplasm. PCR amplification of CMS-specific mitochondrial nucleotide sequences will allow quick and reliable identification of the cytoplasmic types of individual plants at the seedling stage, and assessment of the purity of $F_1$ seed lots. The strategy used in this report for identifying CMS-specific markers could be adopted for many other crops where CMS is used for F1 seed production.

• Journal of Chest Surgery
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• 제33권11호
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• pp.863-868
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• 2000

배경: Cox-Maze III (CM-III) 술식은 복잡한 심방 절개로 인한 긴 수술시간 때문에 다른 개심 수술과 병행하여 시행하기 어려운 단점이 있다. 대상 및 방법: 저자들은 CM-III 술직을 다음과 같이 변형하여 시행하고 그 임상성적을 분석하였다. \circled1 좌심방이를 절제하는 대신 좌심방이를 외부에서 결찰하고, \circled2 폐정맥 분리 절개선과 좌심방이 사이에 냉동절제술을 시행하며, \circled3 우심방이를 절제하는 대신에 우심방 외측 절개선을 우심방이까지 연장하고, \circled4 후종 우심방 절개 하부에서 우심방 외측을 지나 삼첨판막륜으로 향하는 T-자 절개선을 생략하였다. 저자들이 시행한 변형 술식의 용이성과 효율성을 평가하기 위하여, 우리나라에서도 빈도가 높은 류마치스성 승모판막 질환에서, 전통적인 Cox-III 술식(그룹 I)의 임상결과와, 변형된 CM-III 술식(그룹 II)의 임상결과를 비교하였다. 결과: 그룹 I(n=18)에서 동반된 수술은 승모판막 치환술 10례, 승모판막 성형술3례, 승모판막 치환술과 삼첨판막륜 성형술3례, 승모판막 재치환술 2례 등이었다. 그룹II(n=23)에서 동반된 수술은 승모판막 치환술 7례, 승모판막 성형술 5례, 승모판막 치환술과 삼첨판막륜 성형술 1례, 승모판막 재치환술 10례 등이었다. 그룹 I과 그룹 II에서 평균 대동맥 차단 시간(ACC)은 각각 135$\pm$29분과 104$\pm$18 분, 심패바이패스(CPB) 시간은 각각 240$\pm$33분과 185$\pm$42분이었다. 그룹 I과 그룹 II의 평균 추적 관찰 기간은 각각 47$\pm$14 개월과 29$\pm$4 개월이었다. 그룹 I에서는 16례(88.9%)에서 정상 동율동으로 회복되었고 1례에서 심방세동이 남아 있었으며, 다른 1례는 서맥증후군(sick sinus syndrome)으로 인공 심박조율기를 삽입하였다. 그룹 II에서는 21례(91.3%)에서 정상 동율동으로 회복되었고 2례는 심방세동이 지속되었다. 그룹 I에서 정상동율동으로 회복된 16례는 100%(16/16)에서 우심방의 수축을 심장 초음파검사에서 확인할 수 있었으며, 좌심방의 수축은 75%(12/16)에서 확인할 수 있었다. 그룹 II에서는 정상 동율동으로 회복된 21례 중 100%(21/21)에서 우심방의 수축을 확인할 수 있었으며, 좌심방의 수축은 76.2%(16/21)에서 확인할 수 있었다. 결론: 변형 CM-III 술식은 전통 CM-III 술식에 비하여 ACC time(p<0.005)과 CPB time(p<0.001)을 의미있게 줄이면서도 필적할 만 한 정상 동율동 전환율과 심방 수축력의 회복을 보여주었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권8호
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• pp.3447-3450
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• 2015

Objective: To explore the sensitivity of gastric cancer cells to chemotherapy drugs in elderly patients and its correlation with cyclooxygenase-2 (COX-2) expression in cancer tissue. Materials and Methods: Forty-three elderly patients with gastric cancer (observation group) and 31 young patients with gastrointestinal tumors (control group) who were all diagnosed by pathology and underwent surgery in the 89th Hospital of Chinese People's Liberation Army were selected. Drug sensitivity testing of tumor cells in primary culture was carried out in both groups using a methyl thiazolyl tetrazolium (MTT) method, and the expression of COX-2 and the factors related to multi-drug resistance (MDR) in cancer tissue were assessed by immunohistochemistry. Results: The inhibition rates (IR) of vincristine (VCR), 5-fluorouracil (5-FU), oxaliplatin (L-OHP), mitomycin (MMC) and epirubicin (eADM) on tumor cells in the observation group were dramatically lower than in the control group, with statistical significance (P<0.05 or P<0.01). The positive rates of COX-2, glutathione s-transferase-${\pi}$ (GST-${\pi}$) and P glycoprotein (P-gp) expression in cancer tissue in the observation group were all higher than in control group (P<0.05), while that of DNA topoisomerase $II{\alpha}$ ($TopoII{\alpha}$) expression lower than in the control group (P<0.01). In the observation group, COX-2 expression in cancer tissue had a significantly-positive correlation with GST-${\pi}$ and P-gp (r=0.855, P=0.000; r=0.240, P=0.026), but a negative correlation with $TopoII{\alpha}$ (r=-0.328, P=0.002). In the control group, COX-2 expression in cancer tissue was only correlated with P-gp positively (r=0.320, P=0.011). Bivariate correlation analysis displayed that COX-2 expression in cancer tissue in the observation group had a significantly-negative correlation with the IRs of 5-FU, L-OHP, paclitaxel (PTX) and eADM in tumor cells (r=-0.723, P=0.000; r=-0.570, P=0.000; r=-0.919, P=0.000; r=-0.781, P=0.000), but with hydroxycamptothecine (HCPT), VCR and 5-FU in the control group (r=-0.915, P=0.000; r=-0.890, P=0.000; r=-0.949, P=0.000). Conclusions: Gastric cancer cells in elderly patients feature stronger MDR, which may be related to high COX-2 expression.