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題名：以蛋白質體學策略探討高齡衰弱症之血液生物指標
作者：林勁宏
作者(外文)：Ching-Hung Lin
校院名稱：國立體育大學
系所名稱：體育研究所
指導教授：許美智
學位類別：博士
出版日期：2013
主題關鍵詞：發炎；荷爾蒙；肌肉量；肌力；血管收縮素原；激肽原；抗凝血酵素Ⅲ；inflammation；hormone；muscle mass；strength；angiotensinogen；kininogen-1；antithrombin Ⅲ
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高齡者的衰弱情形可能造成個體失能或増加罹病機會，但過去判斷衰弱症，多以觀察外顯的身體活動能力或自我認知回答身體狀況的方式，未有探討個體內在變化的情形，因此如能配合觀察個體生理指標的變化，將可更精確辨別不同的衰弱程度。而肌肉流失是衰弱症的重要表徵，肌肉流失的現象受到體內肌肉流失因子包括發炎反應、胰島素阻抗及合成性荷爾蒙減少的影響，因此本研究以這三個面向，以及身體活動能力與全身肌肉量，探討其於不同衰弱程度的差異性。此外也使用蛋白質體學的策略，分析不同程度之衰弱症者其血清內蛋白質體的差異，同時也探討與身體活動能力、全身肌肉量及肌肉流失因子的關聯性。研究以65歲以上之長者為受試對象，藉由加拿大健康和老化研究之「臨床衰弱量表」中文訪談版問卷，將受試者分為無衰弱症(年齡：77.50.99歲；身高：160.53.13公分；體重：62.53.01公斤)、衰弱症前期(年齡：79.01.00歲；身高：153.34.19公分；體重：65.33.95公斤)、以及衰弱症組(年齡：77.73.83歲；身高：156.67.20公分；體重：62.87.14公斤)，同時增加年輕組別(年齡：27.171.14歲；身高：169.21.92公分；體重：60.173.87公斤)，藉以對照比較年長者各項指標，所有組別男女各為3人。受試者於血液採集前須空腹8小時以上，其後抽取正中肘靜脈血20毫升，以全血及經離心處理後之血清作為分析之用，接續再進行身體組成測量，評估全身肌肉量。血液分析項目包括常規血液生化值、發炎指標、胰島素阻抗、合成性荷爾蒙及蛋白質體學等。資料以獨立樣本單因子變異數分析比較不同衰弱程度各項生物指標的差異，同時以皮爾遜積差相關分析各組身體活動能力與各項生理指標、蛋白質體的關聯性，研究顯著水準訂為=.05。研究結果顯示以問卷篩選出的衰弱症組確實比年輕組、無衰弱症組及衰弱症前期組在握力與6分鐘行走距離明顯較差(13.81+/-1.07 vs 36.72+/-3.53, 26.1+/-3.02 &; 21.64+/-3.78 kg; 215.20+/-42.21 vs 617.3315.50, 438.33+/-42.02 &; 364.33+/-29.56 m)，而年輕組、無衰弱症組與衰弱症前期組之握力明顯與上肢肌肉量有正向關聯性(r=.93, .88 &; .95, p<.05)，反而衰弱症組的肌肉量多寡與其握力無關。且年輕組、無衰弱症組與衰弱症前期組之合成性荷爾蒙與肌肉量及握力皆達顯著正相關，但衰弱症組無任何相關性存在，除此之外衰弱症組發炎指標明顯增加，但與肌肉量及肌力減少無關聯性。藉由蛋白質體學分析，發現衰弱症組之血管收縮素原(angiotensinogen)、激肽原(kininogen-1)及體內抗凝血酵素Ⅲ(antithrombin Ⅲ)表現量明顯高於其他組別(9.85+/-2.04 vs 5.040.85, 4.80+/-0.66 &; 5.50+/-0.92; 26.99+/-1.83 vs 15.30+/-1.51, 15.31+/-1.72 &; 17.94+/-3.14; 23.41+/-1.50 vs 17.95+/-1.43, 9.62+/-0.88 &; 12.96+/-0.69)。本研究主要發現握力與6分鐘行走測驗對衰弱症具有鑑別性，且衰弱症者肌力明顯衰退現象與肌肉流失無關聯。另外高齡者衰弱過程中，體內血管收縮素原增加，伴隨激肽原上升，可能是體內血管功能退化與補償的交互作用機制，以及抗凝血酵素Ⅲ在衰弱過程中的增加，皆與肌力的下降有關，這些可能是衰弱症者體內蛋白質體之重要生物指標。
Elderly frailty may result in disability or increase in the risk of degenerative diseases. The frailty was previously diagnosed by body movement or questionnaire, whereas internal physiological changes can not be determined by using these assessments. If internal physiologic changes can be used to define frailty, more precise status of frailty would be accurately diagnosed. Sarcopenia, which is an important phenomenon of frailty, can be induced by inflammation, insulin resistance, and decrease in anabolic hormones. The purpose of this study was to compare differences in distinct levels of frailty by analyzing sarcopenia, physical activity level, and whole body muscle mass. Proteomics, a new analysis techniques, was also used to investigate various serum proteins in different degrees of frailty. The elderly subjects (n=18; age: yrs; height: cm; weight: kg) were divided to no frailty (NF; n=6; age: 77.50.99 yrs; height: 160.53.13 cm; weight: 62.53.01 kg), prefrailty (PF; n=6; age: 79.01.00 yrs; height: 153.34.19 cm; weight: 65.33.95 kg) and frailty (F; n=6; age: 77.73.83 yrs; height: 156.67.20 cm; weight: 62.87.14 kg) in accordance with the Chinese-Canadian Study of Health and Aging Clinical Frailty Scale. We also recruited a group of young subjects (n=6; age: 27.21.14 yrs; height: 169.21.92 cm; weight: 60.23.87 kg) to serve as the young control group. After an 8 hours fasting, fasting venous blood samples were collected, whole body muscle mass and physical activity ability was measured. The blood analyses included convention biochemical markers, inflammatory markers, insulin sensitivity, anabolic hormones and proteomics. The difference of various biomarkers of each groups were determined by using one-way ANOVA, and the correlations were examined by Pearson product moment correlation. We observed that F group had significantly lower physical activity ability among Y, NF and PF (grip power: 13.81+/-1.07 vs 36.72+/-3.53, 26.1+/-3.02, &; 21.64+/-3.78 kg; 6 min walk test: 215.20+/-42.21 vs 617.33+/-15.50, 438.33+/-42.02, &; 364.33+/-29.56 m). There was a strong positive correlation between muscle strength and muscle mass in NF and PF groups. There was no significant difference in muscle mass loss among experimental groups, but there was significant difference in strength decrease in F group. Although the older subjects without frailty showed decline in the serum level of anabolic hormones, thereby indirectly impairing their physical activity ability. By using proteomics techniques, we observed that the angiotensinogen and kininogen-1 significantly increased during normal process of frailty development (9.85+/-2.04 vs 5.04+/-0.85, 4.80+/-0.66 &; 5.50+/-0.92; 26.99+/-1.83 vs 15.30+/-1.51, 15.31+/-1.72 &; 17.94+/-3.14). This phenomenon might help to explain the possible mechanisms for the interactions between vascular degeneration and compensation, and the relationship between aging and strength decrease. In addition, during normal process of frailty development, antithrombin Ⅲ significantly increased (23.411.50 vs 17.951.43, 9.620.88 &;12.960.69), but muscle strength showed gradual decrease overtime. Our results therefore suggest that antithrombin Ⅲ may serve as an important biomarker during normal process of frailty development.
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