• 제목/요약/키워드: PCR.

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자궁내막증 환자의 정상위치 및 이소성 자궁내막에서의 Osteopontin의 발현 (Expression of Osteopontin in Eutopic and Ectopic Endometrial Tissues in Endometriosis)

  • 구윤희;김정훈;김지선;이영진;김성훈;채희동;강병문
    • Clinical and Experimental Reproductive Medicine
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    • 제34권3호
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    • pp.149-157
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    • 2007
  • 목 적: 본 연구는 세포간 부착과 유착에 관여하는 당단백질의 하나인 osteopontin (OPN)의 mRNA와 단백질의 발현 양상을 자궁내막증 환자의 정상위치 자궁내막조직과 이소성 자궁내막조직, 대조군의 자궁내막조직에서 비교, 분석하기 위해 시행되었다. 연구방법: 개복 또는 복강경 수술을 통하여 자궁내막증으로 확진된 환자 32명을 연구 대상으로 하였고 같은 시기에 자궁경부 상피내암 또는 자궁내막증 이외의 양성 부인과 질환으로 수술적 치료를 받은 환자 34명을 대조군으로 하였다. 수술 시 자궁내막종 또는 복강내 자궁내막 이식물로부터 이소성 자궁내막조직을 얻었고, 동시에 정상위치 자궁내막조직을 생검하였다. 자궁내막조직 내의 OPN mRNA의 발현 정도는 실시간 역전사 중합효소연쇄반응을 이용하여 비교하고 단백질 발현에는 western blot 분석을 사용하였다. 각 군 간의 비교에는 ANOVA와 Krusxal-Wallis test를 사용하였으며 p-value 0.05 미만을 유의한 것으로 판정하였다. 결 과: 월경주기의 증식기와 분비기 모두에서 자궁내막중 환자의 정상위치 및 이소성 자궁내막조직에서의 OPN mRNA의 발현은 대조군의 자궁내막조직에 비해 유의하게 높은 것으로 나타났다 자궁내막중 환자에서 정상위치 자궁내막조직의 경우 OPN mRNA의 발현은 증식기에 비해 분비 기에서 의미 있게 증가하였으나 이소성 자궁내막 조직에서는 분비기에서 뚜렷하게 감소하는 양상으로 나타났다. OPN 단백질의 발현도 mRNA의 발현과 마찬가지로 자궁내막중 환자에서 유의하게 높은 것으로 확인되었다. 결 론: 본 연구의 결과는 자궁내막증이 있는 여성의 정상위치 및 이소성 자궁내막 조직에서의 증가된 OPN mRNA 및 단백질의 발현이 자궁내막 조직의 유착 및 침습에 관여할 수 있음을 시사한다.

Serum Beta-2 Microglobulin: a Possible Marker for Disease Progression in Egyptian Patients with Chronic HCV Related Liver Diseases

  • Ouda, SM;Khairy, AM;Sorour, Ashraf E;Mikhail, Mikhail Nasr
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권17호
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    • pp.7825-7829
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    • 2015
  • Background: Egypt has the highest prevalence of HCV infection in the world (~14.7%). Around 10-15% of HCV-infected persons will advance to cirrhosis within the first 20 years. The incidence of HCC is expected to grow in the next two decades, largely due to HCV related cirrhosis, and detection of HCC at an early stage is critical for a favorable clinical outcome. No simple reliable non-invasive marker has been available till now. B2M, a non-glycosylated polypeptide composed of 99 amino acids, is one of the components of HLA class I molecules on the surfaces of all nucleated cells. It has been reported that the level of serum B2M is elevated in patients with chronic hepatitis C and HCV-related HCC when compared to HCV-negative patients or healthy donors. Determining the clinical utility of serum B2M as a marker for disease progression in Egyptian patients with HCV related chronic hepatitis, cirrhosis and hepatocellular carcinoma was the aim of the present study. Materials and Methods: In this analytical cross sectional study 92 participants were included in 4 equal groups: Group (1) non cirrhotic chronic HCV; Group (2) HCV related liver cirrhosis; Group (3) HCC on top of HCV,; and Group (4) healthy controls. History taking, clinical examination, routine labs and abdominal ultrasound were conducted for all patients, PCR and Metavir scores for group (1) patients, and triphasic CT abdomen and AFP for Group (3) patients. B2M levels were measured in serum with a fully-automated IMX system. Results: The mean serum B2M level of Group (1) was $4.25{\pm}1.48{\mu}g/ml$., Group (2) was $7.48{\pm}3.04$, Group (3) was $6.62{\pm}2.49$ and Group (4) was $1.62{\pm}0.63$. Serum B2M levels were significantly higher in diseased than control group (p<0.01) being significantly higher in cirrhosis ($7.48{\pm}3.04$) and HCC groups ($6.62{\pm}2.49$) than the HCV group ($4.25{\pm}1.48$) (p<0.01). There was a significant correlation between B2M Level and ALK, total and direct bilirubin and INR (p<0.05), and a significant inverse correlation between B2M level and albumin, total proteins, HB andWBCS values (p<0.05). There was no significant correlation between B2M level and viral load or Metavir score, largest tumour size or AFP (p>0.05). The best B2M cut-off for HCV diagnosis was 2.6 with a sensitivity of 100%, a specificity of 92%, a positive predictive value (PPV) of 97% and a negative predictive value (NPV) of 100%. The best B2M cut-off for HCC diagnosis was 4.55 which yielded sensitivity, specificity, positive predictive value, negative predictive values of 74%, 62%, 39.5, 87.8% respectively (p-value <0.01) while best cut-off for cirrhosis was 4.9, with sensitivity 74 % and specificity 74%.The sensitivity for HCC diagnosis increased upon B2M and AFP combined estimation to 91%, specificity to 79%, NPV to 95% and accuracy to 83%. Conclusions: Serum B2M level is elevated in HCV related chronic liver diseases and may be used as a marker for HCV disease progression towards cirrhosis and carcinoma.

Association of Six Susceptibility Loci with Prostate Cancer in Northern Chinese Men

  • Zhang, Yu-Rong;Xu, Yong;Yang, Kuo;Liu, Ming;Wei, Dong;Zhang, Yao-Guang;Shi, Xiao-Hong;Wang, Jian-Ye;Yang, Fan;Wang, Xin;Liang, Si-Ying;Zhao, Cheng-Xiao;Wang, Fei;Chen, Xin;Sun, Liang;Zhu, Xiao-Quan;Zhu, Ling;Yang, Yi-Ge;Tang, Lei;Jiao, Hai-Yan;Huo, Zheng-Hao;Yang, Ze
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6273-6276
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    • 2012
  • Background/Aim: Six prostate cancer (PCa) susceptibility loci were identified in a genome-wide association study (GWAS) in populations of European decent. However, the associations of these 6 single-nucleotide polymorphisms (SNPs) with PCa has remained tobe clarified in men in Northern China. This study aimed to explore the loci associated with PCa risk in a Northern Chinese population. Methods: Blood samples and clinical information of 289 PCa patients and 288 controls from Beijing and Tianjin were collected. All risk SNPs were genotyped using polymerase chain reaction (PCR)-high resolution melting curve technology and gene sequencing. Associations between PCa and clinical covariates (age at diagnosis, prostate-specific antigen [PSA], Gleason score, tumor stage, and level of aggressiveness) and frequencies of alleles and genotypes of these SNPs were analyzed using genetic statistics. Results: Among the candidate SNPs, 11p15 (rs7127900, A) was associated with PCa risk (P = 0.02, odds ratio [OR] = 1.64, 95% confidence interval [CI] = 1.09-2.46). Genotypes showed differences between cases and controls on 11p15 (rs7127900, A), 11q13 (rs7931342, T), and HNF1B (rs4430796, A) (P = 0.03, P = 0.01, and P = 0.04, respectively). The genotype TG on 11q13 (rs7931342, T) was positively associated with an increased Gleason score (P = 0.04, OR = 2.15, 95% CI = 1.02-4.55). Patients carrying TG on 17q24 (rs1859962, G) were negatively associated with an increased body mass index (BMI) (P = 0.03, OR = 0.44, 95% CI = 0.21-0.92) while those with AG on HNF1B (rs4430796, A) were more likely to have PSA increase (P = 0.002). Conclusion: Our study suggests that 11p15 (rs7127900, A) could be a susceptibility locus associated with PCa in Northern Chinese. Genotype TG on 11q13 (rs7931342, T) could be related to an increased Gleason score, AG on HNF1B (rs4430796, A) could be associated with PSA increase, and TG on 17q24 (rs1859962, G) could be negatively associated with an increased BMI in Chinese men with PCa.

Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권18호
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

Disease Progression from Chronic Hepatitis C to Cirrhosis and Hepatocellular Carcinoma is Associated with Increasing DNA Promoter Methylation

  • Zekri, Abd El-Rahman Nabawy;Nassar, Auhood Abdel-Monem;El-Rouby, Mahmoud Nour El-Din;Shousha, Hend Ibrahim;Barakat, Ahmed Barakat;El-Desouky, Eman Desouky;Zayed, Naglaa Ali;Ahmed, Ola Sayed;Youssef, Amira Salah El-Din;Kaseb, Ahmed Omar;El-Aziz, Ashraf Omar Abd;Bahnassy, Abeer Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6721-6726
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    • 2013
  • Background: Changes in DNA methylation patterns are believed to be early events in hepatocarcinogenesis. A better understanding of methylation states and how they correlate with disease progression will aid in finding potential strategies for early detection of HCC. The aim of our study was to analyze the methylation frequency of tumor suppressor genes, P14, P15, and P73, and a mismatch repair gene (O6MGMT) in HCV related chronic liver disease and HCC to identify candidate epigenetic biomarkers for HCC prediction. Materials and Methods: 516 Egyptian patients with HCV-related liver disease were recruited from Kasr Alaini multidisciplinary HCC clinic from April 2010 to January 2012. Subjects were divided into 4 different clinically defined groups - HCC group (n=208), liver cirrhosis group (n=108), chronic hepatitis C group (n=100), and control group (n=100) - to analyze the methylation status of the target genes in patient plasma using EpiTect Methyl qPCR Array technology. Methylation was considered to be hypermethylated if >10% and/or intermediately methylated if >60%. Results: In our series, a significant difference in the hypermethylation status of all studied genes was noted within the different stages of chronic liver disease and ultimately HCC. Hypermethylation of the P14 gene was detected in 100/208 (48.1%), 52/108 (48.1%), 16/100 (16%) and 8/100 (8%) among HCC, liver cirrhosis, chronic hepatitis and control groups, respectively, with a statistically significant difference between the studied groups (p-value 0.008). We also detected P15 hypermethylation in 92/208 (44.2%), 36/108 (33.3%), 20/100 (20%) and 4/100 (4%), respectively (p-value 0.006). In addition, hypermethylation of P73 was detected in 136/208 (65.4%), 72/108 (66.7%), 32/100 (32%) and 4/100 (4%) (p-value <0.001). Also, we detected O6MGMT hypermethylation in 84/208 (40.4%), 60/108 (55.3%), 20/100 (20%) and 4/100 (4%), respectively (p value <0.001. Conclusions: The epigenetic changes observed in this study indicate that HCC tumors exhibit specific DNA methylation signatures with potential clinical applications in diagnosis and prognosis. In addition, methylation frequency could be used to monitor whether a patient with chronic hepatitis C is likely to progress to liver cirrhosis or even HCC. We can conclude that methylation processes are not just early events in hepatocarcinogenesis but accumulate with progression to cancer.

체외수정 생쥐 배아에서의 배아 줄기세포 확립 (Establishment of Mouse Embryonic Stem Cell-like Cells from In Vitro Fertilized Embryos)

  • 문신용;박용빈;김희선;오선경;천대우;서창석;최영민;김정구;이진용;김석현
    • Clinical and Experimental Reproductive Medicine
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    • 제29권1호
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    • pp.1-12
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    • 2002
  • Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.

산양삼(cultivated wild Panax ginseng) 추출물이 조골세포 활성에 미치는 영향 (Effects of Cultivated Wild Panax ginseng Extract on the Proliferation, Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells)

  • 서현주;어현지;김현준;전권석;박광훈;홍세철;정진부
    • 한국자원식물학회지
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    • 제33권4호
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    • pp.227-236
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    • 2020
  • 고령사회에서 노년기 건강의 큰 문제로 대두되고 있는 골다공증은 특히 폐경 후 여성들에게서 가장 그 발생빈도가 높게 나타났으며, 현재 골다공증 예방 및 치료에 사용되고 있는 약제는 대부분 골흡수 억제제로써 진행된 골소실을 회복 시킬 수는 없기 때문에 골형성 증가를 통한 골다공증 예방과 치료에 관한 연구가 활발히 이루어지고 있다. 산양삼(cultivated wild Panax ginseng, CWP)에 대한 연구는 다수가 원기회복, 자양강장 및 면역증강 효과 등에 대한 것이나 골대사에 미치는 영향에 대한 연구는 거의 없는 실정이다. 이에 본 연구에서는 산양삼 추출물이 조골세포에서 골관련 유전자 발현에 미치는 영향을 확인함으로써 골다공증 예방 및 치료 효과를 갖는 천연 소재로의 활용 가능성을 검토하고자 하였다. 산양삼 추출물 처리가 조골 세포의 증식에 미치는 영향을 알아보기 위해 MTT assay를 실시하였고, MC3T3-E1 세포생존률은 FBS가 첨가되지 않은 배양액만 처리한 대조군과 산양삼 추출물을 처리한 실험군 모두에서 동일한 수준으로 나타났으며 이로써 산양삼 추출물의 안전성을 확인할 수 있었다. 또한 산양삼 추출물을 처리한 실험군과 대조군과의 세포증식률을 비교하였을 때 산양삼 추출물 50 ㎍/mL 농도 처리군에서 유의적으로 세포증식이 촉진되었으며 25 ㎍/mL과 100 ㎍/mL 농도 처리군에서도 대조군보다 높은 경향을 나타내었다. 산양삼 추출물이 조골 세포의 활성에 미치는 영향을 알아보기 위해 조골세포의 분화초기 표지인자인 ALP활성을 측정하였으며 그 결과 모든 산양삼 추출물 처리군이 대조군과 비교하여 유의적으로 높은 활성을 나타내었으며 특히 산양삼 추출물 50 ㎍/mL 농도 처리군에서 가장 높은 활성을 나타내었다. 산양삼 추출물의 농도에 따른 석회화 형성도를 확인하기 위해 무기질화된 세포의 기질을 alizarin red로 염색하였고 산양삼 추출물을 처리한 실험군과 대조군과의 석회화 형성도를 비교하였을 때 산양삼 추출물 50 ㎍/mL 농도 처리군에서 유의적으로 석회화 형성이 촉진되었으며 25 ㎍/mL과 100 ㎍/mL 농도 처리군에서도 대조군보다 높은 경향을 나타내었다. 산양삼 추출물이 MC3T3-E1 조골세포에서 골 형성 관련 유전자 발현에 미치는 영향을 확인하기 위해 Runx2, ALP, OPN, OCN등의 유전자를 정량 real-time PCR을 통해 분석하였으며 대조군과 비교하여 모든 산양삼 추출물 처리군에서 농도 의존적이고 유의적으로 골 형성 관련 유전자발현이 증가되었다. 따라서 산양삼추출물이 골 형성 관련 유전자인 Runx2, ALP, OPN, OCN 발현을 증가시켜MC3T3-E1 조골세포의 분화를 촉진하고, 골 석회화 형성 촉진에 기여하였을 것으로 사료된다. 그러나 산양삼 추출물이 골형성과 관련하여 어떠한 기전으로 유전자의 발현을 조절하였는지에 대한 유전자 및 단백질 수준의 추가적인 연구와 산양삼 추출물의 분화 촉진과 석회화 형성능이 산양삼의 사포닌계 진세노사이드 성분의 영향인지에 대한 후속 연구가 필요할 것으로 사료된다.

추출 온도에 따른 금불초 꽃 추출물의 항노화 효능 (Anti-aging Effect of Inula britannica var. chinensis Flower Extract According to the Extraction Temperature)

  • 전지민;유대성;천종우;권순식;전소하;박수남
    • 대한화장품학회지
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    • 제40권1호
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    • pp.109-120
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    • 2014
  • 본 연구에서는 추출 온도에 따른 금불초 꽃(I. britannica var. chinensis) 추출물의 항노화 효능을 알아 보았다. 추출 온도에 따른 세포독성을 살펴본 결과, 상온, $45^{\circ}C$$65^{\circ}C$ 추출물 모두 0.1% 이하의 농도에서 세포독성이 관찰되지 않았다. 0.1% 이하의 농도에서 항노화 효능평가를 수행한 결과, 0.1% 농도의 상온 추출물 경우에 멜라닌 생성 억제율은 24.5%로 $45^{\circ}C$$65^{\circ}C$ 추출물과 비교하여 멜라닌 생성 억제율이 큰 것으로 나타났다. 0.1% 농도의 상온 추출물을 처리한 세포의 히알루론산 합성 관련 유전자 HAS-1, HAS-2 및 HAS-3의 발현율은 각각 123.3%, 137.8% 및 133.2%를 나타냈으며, 이는 비교물질인 L-ascorbic acid (115.6%, 121.0% 및 131.4%) 경우 보다 약간 크게 나타났다. 금불초 꽃 45 $^{\circ}C$ 추출물의 경우, 0.1%의 농도에서 염증유발 유전자인 TNF-${\alpha}$, COX-2 및 IL-1${\alpha}$의 발현율이 대조군 대비 각각 30.3%, 12.8%, 25.7%로 감소하였으며, 이는 상온, $65^{\circ}C$ 추출물과 비교하여 염증유발 유전자 발현 억제 효과가 더 큰 것으로 나타났다. 이상의 결과들로 볼 때, 금불초 꽃 추출물은 추출 온도에 따라 멜라닌 생성 억제, 히알루론산 합성 관련 유전자 발현 증가 효과, 염증유발 유전자 발현 억제 효과를 나타내며, 그로 인하여 피부에서 미백, 보습, 항염 효능을 통해 피부 항노화 효능을 가질 것으로 사료된다. 이는 금불초 추출물이 항노화 화장품 원료로서 응용 가능성이 있음을 시사한다.

Expression of CsRCI2s by NaCl stress reduces water and sodium ion permeation through CsPIP2;1 in Camelina sativa L.

  • Kim, Hyun-Sung;Lim, Hyun-Gyu;Ahn, Sung-Ju
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.194-194
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    • 2017
  • Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

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가뭄 조건 하에서 바이오폴리머 혼합 토양이 Camelina sativa L.의 생장에 미치는 긍정적 영향 (Amended Soil with Biopolymer Positively Affects the Growth of Camelina sativa L. Under Drought Stress)

  • 임현규;김현성;이현숙;신정호;김은석;우효섭;안성주
    • Ecology and Resilient Infrastructure
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    • 제5권3호
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    • pp.163-173
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    • 2018
  • 본 연구에서 사용된 바이오폴리머는 미생물로부터 추출한 ${\beta}$-glucan 및 xanthan gum 이 주성분인 친환경 신소재이며, 토양에 처리하면 강도 증진을 통한 제방의 침식 및 유실 감소뿐만 아니라 토양 내 수분보유력을 증가시키는 것으로 보고되어 있다. 그러나 바이오폴리머가 제방에 적용되었을 때 주변 식생에 미치는 영향에 대한 연구는 미흡한 상황이다. 본 연구는 가뭄 조건에서 토양 내 바이오폴리머 혼합 유무에 따라 Camelina sativa L. (Camelina)의 생육에 미치는 영향을 알아보고자 하였다. 각각 0, 0.25, 0.5, 1%의 바이오폴리머가 혼합된 토양에서 발아로부터 25일간 가뭄 처리 후 표현형을 관찰한 결과, 혼합된 바이오폴리머 농도가 높을수록 가뭄 피해가 감소하였다. 특히 25일 동안의 가뭄 처리된 0.5% 바이오폴리머 혼합구에서 신장, 엽장, 엽폭이 대조구 보다 각각 약 30, 70, 170% 증가하였다. 이후 재급수를 통한 바이오폴리머 혼합구의 회복률 또한 약 80% 높게 나타났다. 식물의 근권 발달을 보기 위해 20일 동안 가뭄을 처리하였으며, 대조구에 비해 바이오폴리머 혼합구에서 측근 형성 및 발달이 약 40% 감소한 것을 확인하였다. Camelina에 가뭄 처리하여 기공전도도, 전해질 유출도, 상대적 수분함량 등 생리적 반응을 조사한 결과, 바이오폴리머의 혼합이 가뭄으로 인한 피해를 각각 약 50, 70, 50% 감소시켰다. 바이오폴리머가 가뭄 조건 하에서 식물의 생장에 미치는 영향을 분자수준에서 알아보기 위해 Reverse Transcription- Polymerase Chain Reaction (RT-PCR)을 통한 유전자 발현양상을 분석하였다. 수분수송 관련 유전자 (aquaporin)인 PIP1;4, 2;1, 2;6 및 TIP1;2, 2;1 발현이 바이오폴리머 혼합구 및 가뭄 처리된 Camelina 잎에서 더 높게 나타났다. 이러한 결과를 종합하였을 때, 바이오폴리머는 가뭄 조건에서 토양 내 수분보유력을 유지시킴으로써 Camelina의 양 수분의 흡수 및 생육에 긍정적인 영향을 주는 것으로 보인다.