| 摘要: | 近年來的研究顯示健康的腸道菌叢，不僅具有健胃整腸的功能，並且有調節免疫及影響中藥吸收、代謝的作用。本研究主要探討腸道菌叢與中醫證型表現關係及腸道菌叢對於中藥代謝的影響。 我們於中國醫藥大學附設醫院過敏性疾病中西醫聯合門診中，篩選48位5至35歲具1年以上過敏性鼻炎病史，並經血液檢查確定對塵蟎過敏的患者，採集糞便檢體進行選擇性培養基細菌培養，同時進行臨床症狀嚴重程度評估及中醫熱證評分，依其熱證評分總分，分成非熱證組(0-5分)及熱證組(6-20分)。 結果顯示過敏性鼻炎的患者中，熱證表現評分與年齡呈現明顯負相關(p < 0.001, r = - 0.46)。熱證表現評分與症狀嚴重度評分具有顯著正相關(p = 0.033, r = 0.31)。熱證表現評分與腸道比菲德氏菌叢數佔腸道中所有菌叢數的比值，雖然未達顯著水準，但呈現有負相關的趨勢(p = 0.06, r = - 0.36)。 此外，我們利用經由毒性試劑tert-butylhydroperoxide, 1.5mM作用之Hep G2細胞株損傷模型進行體外細胞株試驗。試驗分成水飛薊治療組、枸杞子萃取液及枸杞子乳酸菌發酵液組，測量前後的乳酸脫氫酶(lactate dehydrogenase)活性，作為療效評估依據。實驗結果顯示：枸杞子萃取液經乳酸菌發酵後，表現出比原枸杞子萃取液較佳的保護效果 (P < 0.05)。在高效能液相層析儀測定下，進一步發現枸杞子之乳酸菌發酵液比原萃取液在滯流時間(Rt) 5.998之尖峰(peak)上產生了的變化。液相層析指紋圖譜的變化，代表枸杞子萃取液於乳酸菌發酵過程中，改變了枸杞子萃取液原有的成分結構，而加強了枸杞子原有的保護細胞株作用。 本研究的結論顯示：第一，腸道內比菲德氏菌叢數佔腸道中所有菌叢數的比值與過敏性鼻炎患者中醫熱證的表現呈現有顯著的負相關；第二，經乳酸桿菌發酵後之枸杞子發酵液，顯著的增強原萃取液的保護細胞功能，並造成原成分結構的改變。因此腸道菌叢不論是對於中醫證型的影響，或是中藥的代謝可能都佔有重要的角色，值得我們進一步探討腸道菌叢與中醫藥的緊密關係。; Recent researches reveal that good intestinal microflora environment not only helps improve gastrointestinal functions, but it also modulates immune system. Additionally, intestinal microflora promotes absorption and metabolism of Chinese herbal medicine. The aim of the study is to explore the effect of intestinal microflora on Traditional Chinese Medicine (TCM) diagnostic pattern and the metabolism of Chinese herbal medicine. We studied 48 patients who suffered from allergic rhinitis for more than one year from the outpatient services at the Chinese Medical University Hospital. These patients age between 5 to 35 years old, and they were tested to be allergic to Dermatophagoides pteronyssines through blood tests. We collected patients’ stool samples and cultured the bacteria by selective culture mediums. At the same time, we conducted the clinical symptoms severity evaluation score study and TCM heat pattern evaluation score study. The patients were then divided to the “non-heat pattern” group (scoring 0-5) and the “heat pattern group” (scoring 6-20) based on the sum of the TCM heat pattern evaluation scores. The result indicated that in those patients with allergic rhinitis, the TCM heat pattern scores showed significantly negative relationship with age (p < 0.001, r = - 0.46), while showing significantly positive relationship with clinical symptoms severity scores (P = 0.033). In the meantime, heat pattern score showed a tendency of positive relationship with Bifidobacterium percentage in the intestinal microflora (p = 0.06, r = - 0.36). Moreover, we designed an in vitro test with a Hep G2 cell line model injured by toxic reagent, tert-butyl hydroperoxide, 1.5mM. The study was divided into 3 groups: silymarin treat group, Lycium bararum extract treat group and lactic acid bacteria (LAB) fermentation extract of Lycium bararum treat group. The lactate dehydrogenase level before and after treatment was used as the basis to assesses the efficacy of the treatment. The result shows that Lycium barbarum has significant cell protection function (P < 0.05). With the assistance of high performance liquid chromatography (HPLC), we discovered that the peak of retention time (Rt) 5.998 on the finger print of the original Lycium bararum extract disappeared when it was fermented by LAB. The change on the finger print of Lycium barbarum means that the fermentation of the LAB changed the original content of Lycium barbarum and thus enhanced its cell protection function. Finally, we came to two conclusions through the two studies. First, the percentage of Bifidobacterium in the intestinal microflora showed negative relationship with TCM heat pattern score. Second, the fermented Lycium barbarum extract enhance the cell protection effect of the original Lycium barbarum extract. In addition, the fermentation of the LAB changes the original content of Lycium barbarum. The intestinal microflora plays an important role in the TCM pattern representation and the metabolism of C |
