Cisplatin 항암제를 투여한 쥐에서 운동이 근육 대사에 미치는 영향

학위논문(박사) -- 서울대학교대학원 : 사범대학 체육교육과, 2021.8. 송욱.최근 의료기술 발달로 인해 국내 암환자들의 생존율이 높아지고 있다. 이러한 의료기술 중 항암제의 사용률이 지속적으로 증가 되는 추세를 보이고 있다. 특히 Cisplatin이라는 항암제가 많은 암환자들에서 사용 되고 있다. 본 항암제는 알칼리화제에 속하는 항암제로서 주로 고환암, 방광암, 전립선암, 난소암, 두경부암, 폐암, 자궁경부암 치료에 많이 사용되고 있다. 치료의 기전은 세포 내 DNA 내 특정 그룹을 공격하고, 이로 인한 DNA, RNA, 단백질 합성을 저해하여 항암 효과를 보여준다. 하지만, 최근 들어 Cisplatin을 투여 한 암환자들 중에서 여러가지 부작용이 나타남을 보고 하였다. 특히 콩팥독성, 내이독성, 신경독성, 근육감소들이 나타났다. 특히 Cisplatin을 투여한 암환자들 중 80%가 근육 감소를 경험하고 근기능의 저하와 사망률을 높이는 것으로 보고하였다. 현재 임상에서 사용되는 Cisplatin의 용량은 몸무게당 1mg의 용량 이며, 많은 부작용이 있음에도 불구하고 환자들의 치료를 위해 가장 많이 사용되고 있는 실정이다. 많은 선행연구에서는 항암제를 투여한 암환자들에서 근육 감소와 기능 저하를 예방하기 위해 운동이 필요 하다고 보고하였다. 특히 저항성 운동이 항암제를 투여한 암환자들에서 근육대사 증가에 긍정적인 영향을 준다는 것으로 보고 되어졌다. 하지만 최근 연구들에서 항암제가 미토콘드리아의 발생 감소, 에너지 대사 감소, 그리고 자가포식 발현의 감소로 보고 되어졌다. 이러한 감소를 증가 시킬 수 있는 방법은 운동이며, 특히 에너지 대사를 높인 운동 방법이 암예방에 효과가 있는 것으로 보고 하였다. 하지만 항암제를 투요 후 운동의 종류에 따른 미토콘드리아 발생, 근육 감소 대사, 자기포식 변화 비교에 관한 근거들이 부족하다. 따라서 본 논문에서는 Cisplatin을 투여한 Rat모델에서 8주간 운동 종류에 따른 근육 내 발현되는 단백질 종류, 근육 내 조직학적 변화, 단백질 수준에서 에너지 대사, 근육 감소, 자가포식 발현의 변화를 확인하고자 한다. 이를 위해 총 2가지의 연구로 나누어 Cisplatin을 투여한 근육에서 단백질 발현체 분석 및 근육 내 조직학적 변화, 조직내 자가 포식, 에너지 대사, 근육 감소의 단백질 수준 발현 량 조사를 하였다. 첫번째 연구에서는 단백칠체학의 분석을 통해 8주간의 유산소성 운동을 적용한 근육에서 1018개의 단백질 중 손상된 단백질 개선, ubiquitin ligase 관련 발현 증가, 미토콘드리아 ATP합성 관련 단백질 개선 유의미하게 증가가 나타났다. 특히 운동을 하지 않은 그룹에서 ADP-ribose 감소가 증가됨을 확인 하였다. 두번째 연구에서는 cisplatin을 투여한 근육 내 조직학적 변화에서는 운동을 적용한 SOL 근육에서 근육의 단면적 크기, 근육세포의 숫자가 크게 유의미하게 증가 하였다. 근육 조직내 자가포식, 에너지 대사, 근육 감소 관련 단백질의 발현이 운동을 통해 증가 되는 경향을 보였으며, 특히 에너지 대사와 관련 AMPK 및 PGC-1α가 FOXO3a 자가포식 관련 단백질 발현 증가와 관련 있는 것으로 나타났다. 본 연구 결과와 기존의 선행논문들의 보고를 종합해 보면, Cisplatin을 투여 후 운동이 미토콘드리아 발생 증가와 자가포식 관련 인자들의 개선을 통해 근육 감소 예방 효과가 있을 것이다.The survival rate of cancer patients in Korea has been observed to be increasing recently due to the development of cancer-related medical technology. The usage rate of anti-cancer drugs is on an upward trend with these medical technologies. In particular, an anti-cancer drug called Cisplatin is widely used by many cancer patients. This anti-cancer agent is part of a family of alkalizing agents, which are mainly used to treat testicular cancer, bladder cancer, prostate cancer, ovarian cancer, head and neck cancer, lung cancer, and cervical cancer. The mechanism for treatment using Cisplatin has been shown to be through attacking a specific area within the DNA in the cell and then inhibiting the resulting DNA, RNA, and protein synthesis. However, it has been recently reported that there are various side effects in cancer patients who are being treated with cisplatin-related drugs. To date, the most severe observed side effects reported of Cisplatin are: renal toxicity, ototoxicity, neurotoxicity, and muscle loss. In particular, it has been reported that 80% of cancer patients to whom Cisplatin has been administered have experienced muscle wastage and decreased muscle function, and the mortality rate has increased. The current dosage of Cisplatin being used in clinical practice is 1 mg per body weight, and despite its many side effects, it is the first line of treatment for these patients. Many previous studies have reported that exercise is necessary to prevent muscle loss and functional deterioration in cancer patients who underwent chemotherapy. It has also been reported that resistance exercise positively affected muscle metabolism in cancer patients who underwent chemotherapy. Recent studies showed that anticancer drugs have been reported to decrease the generation of mitochondria, decrease energy metabolism, and decrease the expression of autophagy. Moreover, exercise is an effective means of prevention to counteract these issues, resulting in a higher metabolism to address these factors in cancer patients. However, there is insufficient evidence to compare changes in mitochondrial biogenesis, muscle wasting metabolism, and autophagy in relation to different types of exercise after undergoing chemotherapy. Therefore, in this dissertation, over a period of eight weeks, the resistance and aerobic exercise in the cisplatin-administered rat model was used to investigate the expression level of proteins in the proteasome, morphological change in muscles, and altered energy metabolism, muscle atrophy, and autophagy expression. The study was divided into two chapters. These chapters covered protein expression levels using proteomics, morphologically altered muscle, protein expression analysis of autophagy, energy metabolism, and muscle atrophy. In the first study, there were 1018 proteins observed through proteomic analysis. This increased ubiquitin ligase-related expression, repairing the improvement of damaged proteins, and improved mitochondrial ATP synthesis-related proteins significantly increased cisplatin-administered skeletal muscle during the eight weeks of aerobic exercise. In addition, it was confirmed that the decrease in ADP-ribose increased in the group without exercise. In the second study, it was noted that in the morphological changes observed in the muscle treated with Cisplatin, the muscle's cross-sectional area and the number of muscle cells significantly increased in the SOL muscle which had undergone exercise. Furthermore, the expression of proteins related to autophagy, energy metabolism, and muscle loss in muscle tissue showed a tendency to increase through exercise. In particular, AMPK and PGC-1α related to energy metabolism were associated with the increased expression of FOXO3a autophagy-related proteins. Thus, combining the results of this study and reports referenced in previous papers, exercise post-administration of Cisplatin will be effective in preventing muscle loss by increasing mitochondrial generation and improving autophagy-related factors.I. Introduction ７ 1. Study Background ７ A. Current Rates of Using Anticancer Drug in South Korea ７ B. Definition of Cisplatin ８ C. Cisplatin Administration Effect on Skeletal Muscle １０ D. Side Effects of Cisplatin Administration １２ E. The Exercise-Induced Autophagy in Skeletal Muscle １４ F. Autophagy on Mitochondrial Biogenesis and Ubiquitin-Proteasome Proteins in Cisplatin-Administered Skeletal Muscle １７ G. The Effect of Exercise Mitochondrial Alternations in Chemotherapy-Administered Muscle Wasting １７ H. Cisplatin-Administration Affected Cellular Damage, Muscle Cell Death, and Muscle Damage １９ 2. Necessity of Study ２１ 3. Purpose of Study ２３ 4. Hypothesis of Study ２４ II. Materials & Methods ２５ 1. Experimental Design ２５ A. Animal Model & Research Design ２５ B. Exercise Protocols ２８ 2. Experimental Measurements ２９ A. Tissue Collecting ２９ B. Body Weight, SOL & GAS Muscle Weight Measurements ２９ C. Muscle Cross-Sectional Area, Space Area of Muscle, and Number of Myocyte Cells from Hematoxylin & Eosin (H&E) Staining ２９ D. Western Blot from SOL and GAS Cisplatin-Administered Skeletal Muscle ３１ E. Liquid Chromatography – Mass Spectrometry (LC/MS) Analysis ３２ 3. Statistical Analysis ３４ A. Statistical Analysis of Body Weight, Muscle Weight, H&E staining, and Western Blot Results ３４ B. Statistical Analysis of LC/MS Proteomics Results ３４ III. Results ３５ 1. The Results of Morphological-Changed in Cisplatin-Administered Skeletal Muscle ３５ A. The Body Weight, SOL & GAS Muscle Tissue Weight in Cisplatin-Administered Rats ３５ B. The Exercise Affected Muscle Cross-Sectional Area, Space Area, and Number of Myocyte Cells in Cisplatin-Administered Skeletal Muscle ３７ 2. The Results of Western Blot from Cisplatin-Administered Skeletal Muscle ４０ A. Exercise Improved Autophagy-Related Proteins Levels in Cisplatin-Administered Skeletal Muscle ４０ B. Exercise Increased AKT, AMPK, mTOR, and PGC-1α Levels in the Cisplatin-Administered Skeletal Muscle ４３ C. Exercise Regulated AMPK/PGC-1α/FOXO3a Signaling Pathways in Skeletal Muscle of Cisplatin-Administered Rats ４６ D. AMPK/PGC-1a/FOXO3a in Exercise Training Directly Affected Muscle Atrophy as Evidenced by 4EPB1, MuRF 1, and Atrogin-1 Levels ４８ 3. The Results of LC/MS Proteomics Analysis ５１ A. Comparison of Cisplatin-Administered PGC-1α Level between Aerobic Exercise and Cisplatin Control Group ５１ B. Results of Missed Cleavage Rate (%), and Identified Scan Rate (%) in SOL muscle ５５ C. Protein Identification from LC/MS Proteomics ６０ D. Biological Process Results of LC/MS ６３ IV. Discussion ６５ A. The Effect of Exercise Improved Autophagy-Related Proteins in Cisplatin-Administered Skeletal Muscle ６６ B. Cisplatin-Administered Skeletal Muscle with Exercise Preserve Muscle Wasting through Ubiquitin-Proteasome Pathway ６９ C. Cisplatin-Administered Skeletal Muscle Degraded ADP-ribose to Reduce Autophagy-Related Proteins ７２ D. Cisplatin-Administered Skeletal Muscle with Exercise Directly Affected Mitochondrial Biogenesis, which mean AMPK, PGC-1a upregulated FOXO3a ７３ E. Cisplatin-Administered SOL Muscle with Aerobic Exercise Significanlty Affected the Most Effects on Muscle Cross-Sectional Area, Autophagy-Related Factors, and Energy Metabolism ７４ V. Conclusion ７５ VI. Limitations & Future Directions ７７ References ７８ Abstract (Korean) ９５박

A Digital Delay-Locked-Loop for 90 Degree Phase Shift of Memory Receiver Strobe

Maste

Effect of Aerobic Exercise and Its Association with APOE e4 Allele in Cognitive Function of Alzheimers Disease (AD) Patients

학위논문 (석사)-- 서울대학교 대학원 : 체육교육과, 2016. 8. 김연수.Background: Dementia including Alzheimers Disease (AD) with other cognitive function of disorder is increasing in elderly people. The high risk factor of Alzheimers Disease (AD) was genetics of APOE e4. The prevention and treatment are still controversial and the medical industries are trying to develop medications and cognitive functions to dementia people. One of the ways to treat is an exercise that is a non-invasive treatment, in-expensive, lack of side effect, and it decreases the risk factors of dementia with preventing to reduce their cognitive functions. Also, there are lack of studies for exercise program for treatment to dementia. Purpose: Aerobic exercise was more effective way to prevent decline early state of Alzheimers Disease (AD). This study was figured out the effect of aerobic exercise according to APOE e4 presences and how aerobic exercise affect neuropsychological test on between e4 carriers and e4 non-carriers group. Results: The aerobic exercise capacity was significant difference both exercise e4 carriers and e4 non-carrier group compared into control group. This study showed exercise e4 carriers and exercise e4 non-carriers were improved (p = 0.016, r = 0.66) and between exercise e4 carriers and control groups were significant improved (p = 0.007, r = 0.72). Only between exercise e4 carriers group and exercise e4 non-carriers group, both color reading reaction (p = 0.035, r = 0.59) and color reading time per item (p = 0.031, r = 0.605) in CWST were decreased. The COWAT in sematic wrong section was significant improved to decrease between exercise e4 carriers and exercise e4 non-carriers group (p = 0.036, r = 0.627). The DST in backward part also was significant improved scores in exercise e4 carriers (p = 0.039, r = 0.586) and exercise e4 non-carriers (p = 0.011, r = 0.681) than control group. The K-IADL scores was decreased between exercise e4 non-carriers and control group (p = 0.035, r = 0.594) after 12 weeks. Conclusion: There were improved between aerobic exercise and cognitive function in COWAT, CWST, DST, K-IADL after 12 weeks. The APOE e4 carriers also had more effective in COWAT and CWST of cognitive function than exercise e4 non-carriers after aerobic exercise. The DST was effective in the APOE e4 carriers than control. But, the APOE e4 non-carriers was only effective in K-IADL compared to control.I. Introduction 4 1) Significant of the Study 4 2) The purpose of Study 6 3) Research Hypothesis 7 4) Limitations 7 II. Literature Review 8 1) The definition of Alzheimers Disease (AD) and landmark 9 2) APOE genotype and its definition 9 3) The relationship between APOE allele in Alzheimers Disease (AD) and cognitive function 9 4) Timeline of Alzheimers Disease (AD) onset according to age: Lifespan with genetics of Alzheimers Disease (AD) 11 5) Risk factors of Alzheimers Disease (AD) 12 6) Treatment for Dementia and Alzheimers Disease (AD) 14 7) Interaction between physical exercise and APOE e4 allele 15 8) The effect of aerobic exercise and cognitive function within APOE genotype 16 a. The impact of aerobic exercise in APOE allele 16 b. Level of cardiovascular fitness and cognitive function in APOE e4 allele 19 9) The physiological mechanism between effect of aerobic exercise and cognitive function 22 III. Research Method 23 1) Participants 23 2) Sample size 24 3) Study Design 24 4) Exercise Program 25 a. Exericse Intensity 26 b. Exericse Time & Frequency 27 c. Exercise Type 27 5) Measurements 28 a. Neuropsychological Test 28 b. Submaximal Bike Ergometer Test with Maximal Oxygen Uptake (VO2 max) 29 c. Senior Fitness Test (SFT) 31 d. Cardiac Output & Rate-Pressure Prodcut (RPP) 32 e. Tinetti Performance Oriented Mobility Assessment (POMA) Test 34 6) Statistical Diagnosis & Analysis 34 IV. Results 36 1) Baseline of Characteristics 36 2) Difference Aerobic Capacity (VO2 max, Cardiac Output, Rate-Pressure Product) between 3 groups after 12 weeks 38 3) Difference Senior Fitness Test (SFT) between 3 groups after 12 weeks 39 4) Difference Tinetti POMA between 3 groups after 12 weeks 40 5) Difference CWST neuropsychological test between 3 groups after 12 weeks 41 6) Difference COWAT neuropsychological test between 3 groups after 12 weeks 42 7) Difference DSC neuropsychological test between 3 groups after 12 weeks 43 8) Difference TMT neuropsychological test between 3 groups after 12 weeks 43 9) Difference K-MMSE between 3 groups after 12 weeks 44 10) Difference DST between 3 groups after 12 weeks 44 11) Difference BADL between 3 groups after 12 weeks 45 12) Difference K-IADL between 3 groups after 12 weeks 45 V. Discussion 46 1) Aerobic Exercise Capactiy & APOE e4 non- and e4 carriers 46 2) Neuropsychological test differences between APOE e4 non-carriers and e4 carriers after 12 weeks 46 V. Conclusion 47 Bibliography 48 국문 초록 57Maste

Far-End Crosstalk Compensation Circuits for Parallel Microstrip Lines by Using Capacitive-Coupling at Transmitter and Crosstalk-Cancelling Pulse at Receiver

DoctorCircuits are proposed for transmitter and receiver to compensate for the far-end crosstalk (FEXT) of parallel microstrip lines on printed-circuit board (PCB).The transmitter circuit couples the input voltage waveform of an output driver to the output nodes of two adjacent output drivers through a series connection of a capacitor and a compensating driver. The measured bathtub curve shows that the transmitter chip with a 0.13m CMOS process increase the time opening with BER < 1E-12 from 0.04UI to 0.56US at 4.2Gbps.The receiver circuit adds a crosstalk-cancelling pulse to two adjacent input signals to reduce FEXT during the signal transition period, and uses 1-tap decision feedback equalization (DFE) to reduce inter-symbol interference (ISI). The receiver chip, which was applied to a 0.18m CMOS process, increases the maximum data rate with BER < 1E-12 form 2.5Gbps to 3.6Gbps, in a strongly-coupled 2-parallel 2-drom single-ended microstrip SSTL memory channel

A 1V 2.8Gbps 0.18µm CMOS Inverter-Based Digital Cifferential Transmitter with Calibrations of Termination and Misnatch

1

An 11.4mW 3Gbps 0.18mm CMOS Digital Differential Transmitter with Calibrations of Termination and Pre-Driver Mismatch

2

Transition metal-carbon nitride nanotube hybrids catalyst, fabrication method thereof and method for producing hydrogen using the same

본 발명은 질소를 함유하고 있는 탄소나노튜브의 표면에 촉매 활성도가 큰 전이금속을 수 나노크기로 고르게 분산시켜 촉매활성 표면적을 극대화시킨 전이금속-탄소나노튜브 혼성촉매, 그의 제조방법 및 이를 촉매로 이용하여 알칼라인 수소화붕소나트륨(alkaline NaBH4) 용액으로부터 수소를 생산하는 방법에 관한 것으로서, 본 발명에 의한 질소가 첨가된 전이금속-탄소나노튜브 혼성촉매는 연료전지용 수소저장 시스템, 수소자동차용 연료저장 시스템, 전기자동차 및 소형 전자기기의 구동원 등과 같은 수소에너지를 이용한 여러 가지 산업분야에 매우 다양하게 응용될 수 있다

A 1.2V 7-bit 1GS/s CMOS Flash ADC with Cascaded Voting and Offset Calibration

2

A 1V 2.8Gbps 0.18µm CMOS Inverter-Based Digital Cifferential Transmitter with Calibrations of Termination and Misnatch

1