臺灣博碩士論文加值系統

Anstract
Cardiovascular diseases (CVDs) are not only the second cause of death in Taiwan but also the leading cause of death globally, taking an estimation of 17.9 million lives each year. CVDs are collectively a group of heart and blood vessel disorders, including coronary heart, cerebrovascular, rheumatic heart disease, and other conditions. Pro-inflammatory cytokines are important mediators of the immune response which is associated with endothelial dysfunction in CVD patients. Branched fatty acid esters of hydroxy fatty acids (FAHFAs) are a new class of endogenous bioactive lipids that have been found enriched in adipose tissue with inflammation regulatory properties by normalizing TNFα. In the present study, a total of 57 healthy subjects were recruited to determine seven types of FAHFAs and four long-chain fatty acids which are majorly two types of FAHFAs (palmitoleic-acid-9-hydroxy-stearic-acid, 9-POHSA, and oleic-acid-9-hydroxy stearic acid, 9-OAHSA) using Liquid Chromatography mass spectrometry at the baseline characteristics including age, sex, height, body weight, body mass index, etc. in the circulation. The other five types of FAHFAs (5-PAHSA, 9-PAHSA, 12-PAHSA, 9-PAHPA, and 9-SAHSA) were not studied due to the limit of detection or undetected. In this study, we determined higher concentrations of 9-POHSA and 9-OAHSA in healthy human circulation that exhibited anti-inflammatory effects by suppressing LPS stimulated pro-inflammatory cytokines (IL-1β and IL-6) expression in RAW 264.7 cell lines. Additionally, we observed a high concentration of 9-PAHSA (a known anti-inflammatory FAHFA and steroid dexamethasone) along with 9-POHSA and 9-OAHSA which exhibited the relative anti-inflammatory ability. These findings suggest that 9-POHSA, 9-OAHSA, and 9-PAHSA could be inflammatory biomarkers in CVD patients.
Keywords: Fatty acid esters of hydroxy fatty acids (FAHFAs), cardiovascular protection, Biomarkers, Anti-inflammation.

CONTENTS
CHAPTER ONE ......................................................................................................................... 9
1. INTRODUCTION ................................................................................................................... 9
1.1. Cardiovascular disease ..................................................................................................... 9
1.2. Lipids and FAHFAs ....................................................................................................... 10
1.3. Hypothesis ...................................................................................................................... 14
CHAPTER TWO .......................................................................................................................... 15
2. MATERIALS AND METHODS .......................................................................................... 15
2.1. Participants ......................................................................................................................... 15
2.2. Biochemical Analysis ......................................................................................................... 15
2.3. Standards Preparation and Calibration Curves................................................................... 16
2.4. Extraction and Determination of the FAHFAs and Fatty Acids ........................................ 16
2.5. LC-MS Conditions ............................................................................................................. 17
2.6. Cell culture ......................................................................................................................... 18
2.7. RNA Isolation and Quantitative Reverse Transcription-Polymerase Chain Reaction (RT-qPCR) Analysis ......................................................................................................................... 18
2.8. Statistical Analysis ............................................................................................................. 19
CHAPTER TRHEE ...................................................................................................................... 20
3. RESULTS .............................................................................................................................. 20
3.1. Characteristics of the subjects and plasma detection ..................................................... 20
8
3.2. Determination of circulating FAHFAs ........................................................................... 20
3.3. Relationship of FAHFAs with their fatty acids precursor. ............................................ 20
3.4. 9-POHSA, 9-OAHSA, and 9-PAHSA possessed the anti-inflammatory effect ............ 21
3.5. Administration of FAHFAs to RAW 264.7 cells in the absence of LPS ....................... 21
CHAFTER FOUR ......................................................................................................................... 23
4. DISCUSSION AND CONCLUSION ................................................................................... 23
4.1. Discussion ...................................................................................................................... 23
4.2. Conclusion ...................................................................................................................... 24
5. REFERENCE ........................................................................................................................ 25
6. Tables and Figures ................................................................................................................. 27
6.1. Table 1. Baseline characteristics of 57 healthy subjects ................................................ 27
6.2. Table 2. FAHFAS, CV-related biomarkers, and fatty acids levels determined in healthy subjects by using LC-MS .......................................................................................................... 28
6.3. Fig 1. Flow diagram of healthy subject’s enrollment..................................................... 29
6.4. Fig. 2. Determination of FAHFAs ................................................................................ 30
6.5. Fig 3. 9-POHSA and 9-OAHSA were major endogenous FAHFAs in healthy subjects 31
6.6. Fig. 4. The correlation of FAHFAs with their fatty acids precursor ratio ...................... 32
6.7. Fig. 5. Anti-inflammatory effects of 9-POHSA and 9-OAHSA .................................... 33
6.8. Fig. 6. Administration of FAHFAs to RAW 264.7 cells in the absence of LPS. ........... 34

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