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논문 기본 정보

자료유형
학위논문
저자정보

홍남희 (전북대학교, 전북대학교 일반대학원)

지도교수
민지호
발행연도
2013
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전북대학교 논문은 저작권에 의해 보호받습니다.

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이 논문의 연구 히스토리 (2)

2013

물벼룩을 카페인, 아이부프로펜, 아스피린 그리고 테트라 사이클린에 노출시켰을때의 스트레스 반응 분석
홍남희 생물공정공 2013.01 학위논문

2012

물벼룩과 대장균을 이용한 카페인, 아이부프로펜, 아스피린 그리고 테트라사이클린의 위해성 평가
홍남희 ,  레타이황 ,  김양훈 외 1명 환경독성보건학회 심포지엄 및 학술대회 2012.05 학술대회자료

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With increase of the human, necessity of pharmaceuticals used to treat disease increased. However discharged pharmaceuticals have affected aquatic ecosystem. These chemicals have been detected in many countries in sewage treatment plant effluents, surface waters, seawaters, groundwater and some drinking waters. For this reason, pharmaceuticals emerge as environmental contaminants. These pharmaceuticals have a specific mode of action, and some chemicals persist in organism. In this study, extensively consumed caffeine, ibuprofen, aspirin and tetracycline were selected to assess toxicity in aquatic environment.
The effects of pharmaceuticals would be evaluated by analyzing the altered expression patterns of specific responsive genes or proteins as biomarkers. This study carries out RT-PCR and 2-DE to express novel biomarkers in D. magna exposed to pharmaceuticals.
This study consists of four chapters. The First chapter was explained about character of pharmaceuticals and Daphnia magna as aquatic organisms and then toxicity test method was introduced using RT-PCR and 2-DE.
The second chapter, we tested growth rate and gene expression patterns of grpE, recA, katG and fabA from E. coli exposed to pharmaceuticals using RT-PCR. In the result, we are demonstrated that the growth of E. coli is reduced by the pharmaceuticals stress and it showed caffeine, ibuprofen, aspirin and tetracycline affected different pattern to each of genes.
The third chapter, we conducted to examine the effect of these pharmaceuticals on the gene expression level of 5different biomarker in Daphnia magna (i.e. Dhb, Vtg, Arnt, CYP4 and CYP314). For this assay, D. magna were investigated after 48h and 21d exposure to the testing pharmaceuticals. 5 genes have showed different responsive patterns when Daphnia were under stressful conditions
The fourth chapter, we carry out proteome analysis using the 2-DE in D. magna exposed to pharmaceuticals. The results showed that the expression patterns of protein profile in Daphnia magna are changed by toxicity of caffeine, ibuprofen, aspirin and tetracycline. This study found many proteins differently expressed in D. magna in relation to various pharmaceuticals exposure.
In this study, we confirmed not conventional toxicity test but molecular level effect of gene and protein about toxicity of pharmaceuticals. Advantageously this toxicogenomic suggest more progressive than conventional toxicity test to provide novel information by biomarker.
#daphnia magna. pharmaceuticals #toxicity test

목차

TABLE OF CONTENTS Ⅰ
FIGURE LEGENDS Ⅳ
TABLE LEGENDS Ⅶ
ABSTRACT Ⅸ
CHAPTER 1: LITERATURE 1
1.1 CONTAMINATION OF AQUATIC ENVIRONMENT BY PHARMACEUTICALS 1
1.2 DAPHNIA MAGNA 3
1.3 PROTEOMIC APPROACHES IN AQUATIC ECOTOXILOGY 6
1.3.1Reverse Transcription Polymerase chain reaction (RT-PCR) 6
1.3.2 Proteomics analysis by two-dimensional electrophoresis 7
1.4 AIMS OF THE THESIS 8
1.5 THESIS ORGANIZATION 10
CHAPTER2: GENE EXPRESSION OF THE grpE, recA, katG AND fabA IN ESCHERICHIA COLI INDUCED BY TOXICITY OF CAFFEINE, IBUPROFEN, ASPIRIN AND TETRACYCLINE 12
2.1 ABSTRACT 12
2.2 INTROCUTTION 13
2.3 EXPERIMENTAL METHODS 14
2.3.1 Exposure experiments to analysis the gene expression 14
2.3.2 Isolation of RNA samples and Semi-quantitative reverse transcriptase-polymerase chain reaction 15
2.3.3 Statistical analysis 16
2.4 RESULTS AND DISCUSSION 19
2.4.1Growth inhibition of caffeine, ibuprofen, aspirin and tetracycline 19
2.4.2Gene expression of caffeine, ibuprofen, aspirin and tetracycline in E. coli 20
2.5 CONCLUSION 21
CHAPTER 3: ACUTE AND CHRONIC TOXICITY ASSESSMENT AND THEGENE EXPRESSION OF DHB, VTG, ARNT, CYP4 ANDCYP314 IN DAPHNIA MAGNA TO CAFFEINE, IBUPROFEN, ASPIRIN AND TETRACYCLINE 25
3.1 ABSTRACT 25
3.2 INTROCUTTION 27
3.3 EXPERIMENTAL METHODS 33
3.3.1 Daphnia magna culture 33
3.3.2 Acute toxicity test 33
3.3.3 Chronic toxicity test 34
3.3.4 Exposure experiments to analyze the gene expression 35
3.3.5 Isolation of RNA samples and Semi-quantitative reverse transcriptase-polymerase chain reaction 35
3.3.6 Statistical analysis 36
3.4 RESULTS AND DISCUSSION 37
3.4.1 Acute toxicity of caffeine, ibuprofen, aspirin and tetracycline 37
3.4.2 Responses of Dhb, vtg, arnt, cyp4 and cyp314 for acute exposure of caffeine, ibuprofen, aspirin and tetracycline to D. magna 38
3.4.3 Chronic toxicity of caffeine, ibuprofen, aspirin and tetracycline 39
3.4.4 Responses of Dhb, vtg, arnt, cyp4 and cyp314 for chronic exposure of caffeine, ibuprofen, aspirin and tetracycline to D. magna 39
3.5 CONCLUSION 41
CHAPTER 4: PROTEOMIC ANALYSIS OF DAPHNIA MAGNAEXPOSED TO CAFFEINE, IBUPROFEN, ASPIRIN AND TETRACYCLINE 49
4.1 ABSTRACT 49
4.2 INTRODUCTION 50
4.3 MATERIANS AND METHODS 52
4.3.1 Daphnia magna culture 52
4.3.2 Exposure experiments to analyze the protein expression 52
4.3.3 Protein isolation and concentration determination 53
4.3.4 Two-dimensional electrophoresis 54
4.3.5 Silver staining 55
4.3.6 Spots analysis 56
4.4 RESULTS AND DISCUTTION 56
4.4.1 Proteomic analysis of D. magna exposed to caffeine, ibuprofen, aspirin and tetracycline 56
4.5 CONCLUSION 59
CHAPTER 5: CONCLUDING REMARKS 74
REFERENCES 76

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