本文選題：替吉奧 + 腸道菌群�。� 參考：《大連醫(yī)科大學(xué)》2014年碩士論文

【摘要】：替吉奧是一種新型氟尿嘧啶類口服抗癌藥物是抗腫瘤藥替加氟的一種改進(jìn)型制劑，是替加氟、吉美嘧啶、奧替拉西按照摩爾比1：0.4：1組成的復(fù)方制劑[1-3]。用于晚期胃癌的一線治療以及進(jìn)行性或復(fù)發(fā)性頭頸部腫瘤、進(jìn)行性或復(fù)發(fā)性結(jié)腸直腸癌、進(jìn)行性非小細(xì)胞肺癌、轉(zhuǎn)移性乳腺癌、轉(zhuǎn)移性胰腺癌和膽管癌等的治療[4]，據(jù)統(tǒng)計(jì)日本晚期胃癌的治療，有80%以上的病例使用替吉奧，治療有效率（CR+PR）可達(dá)44.6%。替吉奧中奧特拉西能夠降低5-氟尿嘧啶在腸組織內(nèi)產(chǎn)生磷酸化(磷酸化的結(jié)果是導(dǎo)致胃腸道不良反應(yīng)的主要原因)，但其胃腸反應(yīng)仍然不容忽視，多表現(xiàn)為便秘、腹瀉及胃腸道出血，加重患者的痛苦，不利于病情的好轉(zhuǎn)。 微生態(tài)學(xué)是一門研究生物體正常微生物群與其宿主相互依賴、相互制約規(guī)律的科學(xué)。我國(guó)著名微生態(tài)學(xué)專家康白教授將微生態(tài)系統(tǒng)的定義為：微生態(tài)系統(tǒng)是指在一定結(jié)構(gòu)的空間內(nèi)，正常微生物群以其宿主的組織和細(xì)胞及其代謝產(chǎn)物為環(huán)境，在長(zhǎng)期的進(jìn)化過(guò)程中形成的能獨(dú)立進(jìn)行物質(zhì)、能量及信息相互交流的統(tǒng)一的生物系統(tǒng)。腸道微生態(tài)是人體最大的微生態(tài)系統(tǒng)，被認(rèn)為是人體一個(gè)重要的器官，參與人體的新陳代謝，對(duì)人體健康起著至關(guān)重要的作用。腸道菌群處于健康狀態(tài)時(shí)，維持著人體的的各種功能，其機(jī)制可總結(jié)為以下幾點(diǎn)：營(yíng)養(yǎng)促進(jìn)作用、參與腸粘膜的增生、免疫作用等。當(dāng)腸道菌群失衡時(shí)，致病菌就會(huì)大量滋生，導(dǎo)致腸粘膜粗糙、血液流通不暢、產(chǎn)生大量有害物質(zhì)、胃腸蠕動(dòng)異常以及導(dǎo)致病原體的侵入。現(xiàn)已證明腸道菌群失衡與肥胖、高血脂、胃炎、癌癥等多種疾病有關(guān)。 近一個(gè)世紀(jì)來(lái)，人們對(duì)益生菌的研究越來(lái)越多，越來(lái)越深入，研究發(fā)現(xiàn)益生菌對(duì)人體健康有益。益生菌能夠產(chǎn)生β-半乳糖苷酶催化乳糖分解，減輕乳糖不耐癥，還能促進(jìn)蛋白質(zhì)和鈣等營(yíng)養(yǎng)物質(zhì)的吸收，部分益生菌能夠合成維生素B12和葉酸，起到營(yíng)養(yǎng)機(jī)體的作用。益生菌在繁殖過(guò)程中產(chǎn)生大量的有機(jī)酸，刺激腸壁蠕動(dòng)，增加胃動(dòng)力。另外益生菌定植后，能夠在腸粘膜上形成一層保護(hù)膜，，起到屏障作用，拮抗致病菌。益生菌能夠產(chǎn)生細(xì)菌素、抗生素及其他代謝物、空間爭(zhēng)奪等抑制致病菌的增值，維持腸道微生態(tài)的平衡，保持宿主正常的營(yíng)養(yǎng)、免疫、消化等生理功能[5]。 對(duì)于腸道菌群的檢測(cè)有多種方法，依靠體外培養(yǎng)的技術(shù)例如細(xì)菌涂片是最直接、最簡(jiǎn)單、快速的經(jīng)典方法，區(qū)分革蘭氏陽(yáng)性菌與革蘭氏陰性菌的比例，可以初步判斷腸道菌群是否失衡；還有細(xì)菌培養(yǎng)法，它是一種定量的細(xì)菌培養(yǎng)法。雖然體外培養(yǎng)技術(shù)簡(jiǎn)單易操作，但可培養(yǎng)細(xì)菌的比例與腸道微生態(tài)細(xì)菌總量相比，存在較大差異，因?yàn)楹芏嗉?xì)菌無(wú)法實(shí)現(xiàn)體外培養(yǎng)，可培養(yǎng)的細(xì)菌只占總數(shù)的30%-40%，無(wú)法完全反應(yīng)腸道菌群的組成，而且體外培養(yǎng)環(huán)境與腸道環(huán)境不能完全相同加之人為操作因素，也可能會(huì)改變?cè)械奈⑸锶郝�，隨著科技的發(fā)展分子生物技術(shù)開(kāi)始應(yīng)用于腸道菌群的檢測(cè)，rRNA分子在生物進(jìn)化過(guò)程中始終保持相對(duì)恒定的生物學(xué)功能和保守的堿基排列序列，同時(shí)也存在為保守區(qū)和可變區(qū)，研究rRNA基因序列可以發(fā)現(xiàn)各物種間的體統(tǒng)發(fā)生關(guān)系，其中16sRNA長(zhǎng)度約1540bp，較易于進(jìn)行序列的測(cè)定，近年來(lái)以16sRNA為基礎(chǔ)的分子生物學(xué)技術(shù)開(kāi)始應(yīng)用于腸道微生態(tài)的分析，包括實(shí)時(shí)定量PCR、熒光原位雜交、核酸探針技術(shù)、TGGE、PCR-DGGE、TGGE、T-RFLP等[6]。其中PCR-DGGE對(duì)菌群種屬進(jìn)行定性、半定量分析，其優(yōu)點(diǎn)是可將相同長(zhǎng)度不同序列的PCR擴(kuò)增片段進(jìn)行分離，檢測(cè)率高、重復(fù)性好。DGGE圖譜中，條帶比較粗亮說(shuō)明細(xì)菌密度大，條帶細(xì)淺則菌密度小，另外DGGE圖譜中條帶的數(shù)量也體現(xiàn)的腸道菌群的種群數(shù)量，因此在后面對(duì)DGGE圖譜的分析中我們也會(huì)利用相應(yīng)軟件分析條帶的灰度值及數(shù)量[7]。 本實(shí)驗(yàn)用357F-GC／518R引物擴(kuò)增16S rDNA的V3可變區(qū),基于16S rRNA基因的聚合酶鏈?zhǔn)椒磻?yīng)-變性梯度凝膠電泳(PCR-DGGE)技術(shù)應(yīng)用于腸道微生態(tài)學(xué)，對(duì)DGGE指紋圖譜進(jìn)行相似性、多樣性分析及優(yōu)勢(shì)條帶的序列分析，檢測(cè)替吉奧對(duì)人體腸道菌群的影響，以及兩歧雙歧桿菌、嗜酸乳桿菌、嬰兒雙歧桿菌組成的益生菌制劑能否修復(fù)由替吉奧引起的腸道菌群失衡,為臨床合理用藥提供參考。
[Abstract]:Tegio is a new type of oral anticancer drug, a new type of antitumor drug tegafur. It is a first-line treatment of advanced gastric cancer and progressive or recurrent head and neck tumors by [1-3]., a compound preparation of tegafur, gimilacil, and oretirine, based on the mole ratio 1:0.4:1, with progressive or recurrent head and neck tumors. Colorectal cancer, progressive non-small cell lung cancer, metastatic breast cancer, metastatic pancreatic cancer and cholangiocarcinoma are treated with [4]. According to the treatment of advanced gastric cancer in Japan, more than 80% of the cases were used as teggio, and the therapeutic efficiency (CR+PR) could reach 44.6%. teggio in oteasi can reduce the phosphorylation of 5- fluorouracil in the intestinal tissue (phosphorus) The result of acidification is the main cause of the adverse reaction of the gastrointestinal tract, but the gastrointestinal reaction still can not be ignored. It is often manifested in constipation, diarrhea and gastrointestinal bleeding, which aggravates the suffering of the patients and is not conducive to the improvement of the condition.
Microbiology is a science that studies the mutual dependence and restriction of the normal microorganism group and its host. Professor Kang Bai, a famous microecology expert in China, defines the microecosystem as: the microecological system refers to the tissues and cells and their metabolites of the normal microorganism in a certain structure. For the environment, a unified biological system is formed in the process of long-term evolution, which can independently carry out material, energy and information. The intestinal microecology is the largest microecosystem in the human body. It is considered to be an important organ of the human body. It plays a vital role in human body health. In a healthy state, it maintains various functions of the human body. Its mechanism can be summarized as following: nutrition promotion, participation in intestinal mucosa hyperplasia, immune function and so on. When the intestinal flora is unbalanced, the pathogenic bacteria will grow in large numbers, causing the rough intestinal mucosa, poor circulation of blood, producing a large number of harmful substances, abnormal gastrointestinal peristalsis and causing disease. The invasion of cancer has proved that intestinal flora imbalance is related to obesity, hyperlipidemia, gastritis, cancer and many other diseases.
In the past century, more and more studies have been made on probiotics. Research has found that probiotics are beneficial to human health. Probiotics can produce beta galactosidase to catalyze lactose decomposition, reduce lactose intolerance, and promote the absorption of nutrients such as protein and calcium. Some probiotics can synthesize vitamin B12 and folic acid. Probiotics produce a large amount of organic acids in the breeding process, stimulate the peristalsis of the intestinal wall, and increase the motility of the stomach. In addition, probiotics can form a protective membrane on the intestinal mucosa after probiotic colonization. It can act as a barrier and antagonize pathogenic bacteria. Probiotics can produce bacterial, antibiotics and other metabolites, space contention and so on. The increment of pathogenic bacteria can maintain the balance of intestinal microecology and maintain the normal physiological functions such as nutrition, immunity and digestion of the host. [5].
There are many methods for the detection of intestinal flora. In vitro culture, such as bacterial smear is the most direct, simplest and fast classical method to distinguish the proportion of Gram-positive bacteria and Gram-negative bacteria, it can preliminarily judge whether the intestinal flora is unbalance, and the bacterial culture method is a quantitative bacterial culture method. In vitro culture technology is simple and easy to operate, but the proportion of the bacteria can be compared with the total amount of intestinal microecological bacteria, because many bacteria can not be cultured in vitro. The bacteria can only account for 30%-40% of the total number of bacteria, which can not fully reflect the composition of intestinal flora, and the culture environment in vitro and the intestinal environment can not be exactly the same. In addition to human manipulation factors, it may also change the original microbial community. With the development of technology, molecular biotechnology has been applied to the detection of intestinal microflora. RRNA has maintained relatively constant biological function and conservative sequence sequence throughout the process of biological evolution, and also exists as a conservative area and variable region, and studies rR NA gene sequence can find the relationship between species, in which the length of 16sRNA is about 1540bp, and it is easier to determine the sequence. In recent years, the molecular biology technology based on 16sRNA has been applied to the analysis of intestinal microecology, including real-time quantitative PCR, fluorescence in situ hybridization, nucleic acid probe technology, TGGE, PCR-DGGE, TGGE, T-RFLP and so on. In 6]., PCR-DGGE was qualitative and semi quantitative analysis. The advantage was that the PCR amplification fragments of the same length and different sequences could be separated, the detection rate was high and the reproducibility good.DGGE atlas. The strip was bright and bright indicating that the density of bacteria was large, the strip was small and the density of the bacteria was small. In addition, the number of bands in the DGGE map also reflected the intestinal bacteria The number of groups, so in the subsequent analysis of the DGGE map, we will also use the corresponding software to analyze the gray value and the number of bands [7].
In this experiment, the V3 variable region of 16S rDNA was amplified with 357F-GC / 518R primers, and the polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) based on 16S rRNA gene was applied to the intestinal microecology. The similarity, diversity analysis and sequence analysis of the dominant bands were used to detect the shadow of the intestinal flora of the human body. Whether or not the probiotics of Bifidobacterium bifidus, Lactobacillus acidophilus and Bifidobacterium infantile can repair the imbalance of intestinal flora caused by tegio, and provide a reference for clinical rational use.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 王文建;鄭躍杰;羅宏英;;嬰兒牛奶過(guò)敏23例臨床分析[J];中國(guó)兒童保健雜志;2011年04期

2 吳瑜,易成,王樹(shù)人,張敏,張靜;嬰兒雙歧桿菌對(duì)小鼠黑色素瘤模型腫瘤組織的靶向性[J];四川大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2003年03期

3 盧亞亞;莫清萍;張巧玲;張泉山;丘小汕;;雙歧桿菌對(duì)卵清蛋白致敏大鼠腸道黏膜屏障的保護(hù)作用[J];臨床兒科雜志;2009年09期

4 姜彬言;王巧民;胡樂(lè)義;;腸易激綜合征患者腸道菌群分析及應(yīng)用益生菌療效觀察[J];中國(guó)臨床保健雜志;2012年02期

5 李軍訓(xùn);羅學(xué)剛;高潔;郝冉;張同存;;益生菌的分類、生理功能與有效性評(píng)價(jià)研究進(jìn)展[J];中國(guó)農(nóng)業(yè)科技導(dǎo)報(bào);2010年06期

6 許亮;許學(xué)新;;益生菌在治療難治性幽門螺桿菌感染中的價(jià)值[J];臨床消化病雜志;2012年04期

7 楊文方;李旭;王翔;王寶珠;楊柳;;陰道微生態(tài)的臨床研究及意義[J];實(shí)用婦產(chǎn)科雜志;2012年03期

8 唐文靜;黃娟;鐘燕;周侃;陳兵;;益生菌干預(yù)對(duì)卵清蛋白過(guò)敏幼鼠模型的免疫調(diào)節(jié)作用[J];上海交通大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2010年01期

9 張澤生;王俊輝;李健;趙春曉;;四株益生菌對(duì)高脂血癥倉(cāng)鼠降血脂作用的研究[J];食品工業(yè)科技;2012年12期

10 丁淑賢;張昌k(;;金雙歧輔治嬰幼兒濕疹40例報(bào)告[J];咸寧學(xué)院學(xué)報(bào)(醫(yī)學(xué)版);2007年05期

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