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研究生:尤顥宇
研究生(外文):Hao-Yu Yu
論文名稱:腸道微生物菌相與肌肉痙攣相關性研究
論文名稱(外文):Study for the correlation of gut microbiota and spasticity
指導教授:林槐庭林槐庭引用關係
指導教授(外文):Hwai-Ting Lin
口試委員:陳姝希,吳汶蘭
口試委員(外文):Su-Zy Chen,Wen-Lan Wu
口試日期:2020-01-15
學位類別:碩士
校院名稱:高雄醫學大學
系所名稱:運動醫學系碩士在職專班
學門:醫藥衛生學門
學類:復健醫學學類
論文種類:學術論文
論文出版年:2020
畢業學年度:108
論文頁數:77
中文關鍵詞:痙攣腸道微生物菌相次世代定序菌門相對豐度F/B比值
外文關鍵詞:spasticitygut microbiotanext generation sequencingrelative abundance of phylumF/B ratio
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人體有許多種微生物群聚於身體各處,正常情形下兩者互利共生、和平共存並共同演化。在人體消化道的腸道微生物估計與人體細胞數目相當,其多樣的微生物種類組成腸道微生物菌相 (gut microbiota)。腸道微生菌的代謝體能透過腸道上皮組織,由迷走神經傳導至中樞神經系統,進而對大腦產生影響,又稱微生物-腸-腦軸線(microbes-gut-brain axis)。而肌肉痙攣是造成病人運動障礙的主要原因,若無積極治療與復健,可能終身癱在床上,倚靠他人照顧生活,造成照護者的長期壓力與負擔。
目前研究對於腸道微生物菌相與肌肉痙攣的關聯性尚不清楚,也無法提供改善肌肉痙攣的飲食調整方案。因此本研究的目標是藉由橫斷面的研究,探討護家臥床肌肉痙攣住民管灌果汁與否及住民狀況差異,是否會有不同的腸道微生物菌相,以及不同的肌肉痙攣程度。
本試驗收案對象為年滿60歲在本院護理之家照護3個月以上的住院患者,且經物理治療師評估為具有肌肉痙攣症狀。受試者以 Modified Ashworth Scale評估為肌肉痙攣患者後,再進行性別、病因、住院時間等紀錄。於紀錄完成48小時內收集受試者糞便,以次世代定序法(next-generation sequencing)檢測糞便內的微生物總基因體(metagenomics),進行腸道微生物菌相分析及物種多樣性 (diversity)與豐度 (abundance) 注釋。統計方法以相關性分析檢視肌肉痙攣分數與物種多樣性指標(Shannon index 與 Simpson index),以及厚壁菌門(Firmicutes)與擬桿菌門(Bacteroidetes)比值(F/B ratio)的相關性。本研究使用Mann Whitney檢定探討受試者在不同性別、病因、住院時間長短(三年以上或是三年以下)與服用果汁與否等因子對於其肌肉痙攣、菌門相對豐度與多樣性是否有所差異。p<0.05 視為具統計差異。
結果顯示受試者不同性別、病因、住院時間長短其肌肉痙攣程度並無顯著差異;果汁服用與否其肌肉痙攣程度有顯著差異,p值為0.02。Bacteroidetes之相對豐度在無攝取果汁組有21%,攝取果汁組則是34%顯著較多,p值為0.03,power值為0.34。不同性別、病因、住院時間與果汁服用與否其alpha多樣性皆無顯著差異。肢體痙攣度與腸道微生物菌相的相關性則是發現肌肉痙攣度與Firmicutes(r=0.44)、F/B比值(r=0.44)呈現中度正相關。
肢體痙攣度與腸道微生物菌相雖然無達到顯著相關。長期臥床的住民因肢體痙攣而產生關節角度限制或肌肉攣縮等症狀,造成晚年臥床生活品質不佳,照顧護理方面的困難,有可能是因為腸道微生物菌相的改變而造成肢體痙攣。根據本研究數據及相關文獻探討證據顯示,護理之家住民性別、中風、住院時間、單一化飲食有否供應果汁的差別比較之下,餵食果汁在住民痙攣程度上有顯著差異,肢體痙攣程度卻與腸道內的Firmicutes和Bacteroidetes的豐度比例中度相關和Bacteroidetes的豐度也因為果汁餵食而呈現顯著差異。因此護理之家住民的肢體痙攣情形和其體內的腸道微生物菌相也有一些關聯,但因樣本數不足關係無法達到統計上的差異,但發現適量讓痙攣的臥床病人攝取果汁中的膳食纖維,有助於改善減輕痙攣情形。
There are many kinds of microbiome in the human body. Under normal circumstances, the two mutually benefit each other, and they coexist and co-evolve peacefully in their lifetime. The gut microbiota in the human digestive tract are estimated to be comparable to the number of human cells, and their diverse microbial species constitute the gut microbiota. Metabolites of gut microbiota can pass through the intestinal epithelial tissue and are transmitted from the vagus nerve to the central nervous system, which in turn affects the brain, also known as the microbes-gut-brain axis. In addition, spasticity is the main cause of dyskinesia in patients. Without active treatment and rehabilitation, the patients may stay in bed for the rest of your life and rely on others to take care of their life, causing long-term pressure and burden on caregivers.
The subjects recruited in this study were hospitalized patients who had been nursing at the nursing home of the hospital for more than 3 months at the age of 60, and were assessed by the physical therapist whether having muscle spasticity. Subjects were evaluated their muscle spasticity level by Modified Ashworth Scale and then recorded their gender, etiology, and length of hospital stay. The feces of the subjects were collected within 48 hours of the completion of the recording, and the microbial genomes in the feces were detected by next-generation sequencing for analysis of the gut microbiota. Make a note on species diversity and abundance. Statistical methods were used to examine the correlation between spasticity score and species diversity index (Shannon index and Simpson index), and the ratio of Firmicutes to Bacteroidetes (F/B ratio). The study also using Mann Whitney test to find out whether gender, etiology, length of stay, and drink juice groups would have differences in their spasticity, relative abundance and diversity. p<0.05 was considered statistically significant different.
The results found that there was no significant difference in the effects of spasticity grouping on groups of gender, etiology, hospitalization time, however, the sum of limb muscle spasticity by juice was significant less than non-juice group with p-value 0.02. The relative abundance of Bacteroidetes was 21% in non-juice group was also significant less than juice group which has 34% (p-value=0.03). There was no significant difference in the effect of alpha diversity according to different gender, etiology, hospitalization time, and juice. There were medium positive correlations between the spasticity and Firmicutes (r=0.44), F/B ratio (r=0.44).
Although the consequence was that llimb spasticity was not significantly correlated to gut microbiota due to the smaller sample size, residents who had been bedridden for a long time had symptoms such as joint angle limitation or muscle contracture due to spasticity, resulting in poor quality of life in bed in their later years. Difficulties in care might be caused by changes in gut microbiota. According to the data of this study and relevant literature, the evidence showed that there was no difference in the degree of limb spasticity and the alpha diversity by gender, etiology, hospitalization time; however, the degree of limb spasticity was significantly different on juice group. Since the degree of limb spasticity was medium correlated to the abundance ratio of Firmicutes and Bacteroidetes in the intestine, the relative abundance of Bacteroidetes was significantly different by feeding juice. Therefore, we could not say the spasticity of the nursing home residents was totally not correlated to the gut microbiota as not reaching significantly different statically due to small sample size. It was happier to be found by feeding the juice the bed-ridden patient in the nursing home could decrease the spasticity for the diet fiber.
目錄
中文摘要 I
ABSTRACT IV
表目錄 XI
圖目錄 XII
第一章 緒論 1
第一節 研究背景 1
第二節 研究目的 4
第二章 文獻探討 5
第一節 肌肉痙攣 5
第二節 腸道微生物菌相 10
第三章 研究方法 24
第一節 實驗設計及流程 24
第二節 受試者募集(含收案條件、排除條件) 26
第三節 肌肉痙攣評估 27
第四節 糞便收集方式 29
第五節 腸道微生物菌相分析 30
第六節 統計方法 32
第四章 資料分析與結果 33
第一節 性別、病因、入住時間、果汁分組中肌肉痙攣的差異性 34
第二節 不同性別、病因、入住時間、果汁在菌門豐度的差異性 36
第三節 不同性別、病因、入住時間、果汁攝取其ALPHA多樣性的差異性 40
第四節 肢體痙攣度與腸道微生物菌相的相關性 44
第五章 討論 50
第一節 性別、中風、入住時間、果汁分組中肌肉痙攣的差異性 51
第二節 不同性別、中風、入住時間、果汁其菌門豐度的差異性 53
第三節 不同性別、中風、入住時間、果汁其ALPHA多樣性的差異性 54
第四節 肢體痙攣度與腸道微生物菌相的相關性 55
第五節 研究限制 57
第六章 結論 58
參考文獻 59
附錄 75
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