本文選題：侵襲性真菌病 + 真菌　； 參考：《復(fù)旦大學(xué)》2013年碩士論文

【摘要】：目的： 證實應(yīng)用廣譜抗生素和(或)應(yīng)激狀態(tài)可致腸道菌群紊亂,導(dǎo)致原本少量的正常菌群(如白色念珠菌)及過路菌大量增殖,證實急性應(yīng)激狀態(tài)下腸道屏障在侵襲性真菌病發(fā)病過程中的作用。 方法： 本課題為動物實驗,實驗對象是Wistar大鼠,將60只大鼠分為三組,雌雄不記,每組20只,第一組為空白對照組；第二組為抗生素組,每日靜脈給予頭孢曲松(Ceftriaxone, CTRX)100mg/kg一次[16][17]第三組為抗生素+胰腺炎組,腹腔注射20%精氨酸(L-Arginine,L-ARG)1.25mg/kg,兩次,中間間隔一小時,造成急性出血性胰腺炎模型,同樣給予頭孢曲松,劑量及方法同第二組。其余飲食、飲水、鼠籠等實驗條件相同。分別于實驗開始第7天、第14天,各取每組10只小鼠,離斷頸椎后無菌操作環(huán)境下快速取心內(nèi)血、門靜脈血、腸內(nèi)容物(Intestinal Content,IC)、腸壁及腸系膜淋巴結(jié)。取血、組織勻漿(Tissue Homogenate,TH)及腸內(nèi)容物做真菌培養(yǎng),取腸壁組織及胰腺做病理鏡檢。若同一實驗動物有兩個或兩個以上樣本培養(yǎng)結(jié)果陽性,則分離真菌,應(yīng)用18S rDNA內(nèi)轉(zhuǎn)錄間隔區(qū)(Internal Transcribed Spacer, ITS)序列分析技術(shù),抽提基因組DNA,電泳檢測,PCR擴(kuò)增,電泳檢測,切膠純化測序,拼接序列,分析序列結(jié)果,達(dá)到菌株層面的鑒定,確定是否有真菌突破感染。 結(jié)果： 1.在實驗第7天,抗生素組及抗生素+胰腺炎組動物的腸內(nèi)容物真菌培養(yǎng)陽性結(jié)果與空白組對比有顯著差異,表明實驗第7天已存在菌群紊亂,真菌增殖。 2.在實驗第14天,抗生素組及抗生素+胰腺炎組動物的腸內(nèi)容物真菌培養(yǎng)陽性結(jié)果與空白組對比有顯著差異,表明實驗第14天仍存在菌群紊亂,真菌增殖。 3.抗生素組與抗生素+胰腺炎組動物的腸內(nèi)容物真菌培養(yǎng)陽性結(jié)果比較,無論是在實驗第7天還是在實驗第14天,兩者均無統(tǒng)計學(xué)差異,表明長時間應(yīng)用抗生素的基礎(chǔ)上,負(fù)荷應(yīng)激因素,并不會加重菌群紊亂。 4.在實驗第7天及實驗第14天,抗生素組及抗生素+胰腺炎組與空白組相比,無明顯突破腸道屏障證據(jù),未造成侵襲性真菌感染。 5.腸道屏障的免疫功能在其防止侵襲性真菌病的發(fā)生過程中有重要作用。 結(jié)論： 長期應(yīng)用廣譜抗生素可造成腸道菌群紊亂,真菌增殖,隨著抗生素應(yīng)用時間的延長,負(fù)荷應(yīng)激因素,并不會加劇菌群紊亂；在真菌成為腸道優(yōu)勢菌群的情況下,腸道固有屏障的保護(hù)作用,仍可保證急性應(yīng)激情況下,真菌不能突破腸道屏障,造成侵襲性真菌病,腸道屏障的免疫功能在其中可能起重要作用。
[Abstract]:Objective: It was proved that the use of broad-spectrum antibiotics and / or stress could lead to intestinal flora disorder, resulting in the proliferation of a small number of normal flora (E. g., Candida albicans) and passageway bacteria. The role of intestinal barrier in the pathogenesis of invasive mycosis was confirmed under acute stress. Methods: This subject is an animal experiment, the experimental object is Wistar rats, 60 rats are divided into three groups, male and female, 20 rats in each group, the first group is a blank control group, the second group is an antibiotic group, Ceftriaxone was given intravenously daily and CTRX)100mg/kg was given once [16] [17] the third group was treated with antibiotic pancreatitis, 20% arginine L-Arginine L-ARGN 1.25mg / kg intraperitoneally, twice, with an interval of one hour, to make the model of acute hemorrhagic pancreatitis, and also to give ceftriaxone, the same dose of ceftriaxone. The dose and method were the same as those in the second group. The rest of the diet, drinking water, squirrel cage and other experimental conditions are the same. On the 7th and 14th day of the experiment, 10 mice in each group were taken from each group. The blood of heart, portal vein, intestinal contents, intestinal wall and mesenteric lymph nodes were collected quickly in aseptic environment after cervical amputation. Blood, tissue homogenate, tissue homogenate (THH) and intestinal contents were used for fungal culture, and intestinal wall tissue and pancreas were taken for histopathological examination. If two or more samples of the same laboratory animals were positive, fungi were isolated. Genomic DNAs were extracted by internal Transcribed Spacer, ITS) sequence analysis technique of 18s rDNA internal transcribed spacer, and then amplified by electrophoresis, purified and sequenced by gumming. Splicing the sequence and analyzing the sequence results to identify the strain level to determine whether there is a fungal breakthrough infection. Results: 1. On the 7th day of the experiment, the positive results of fungal culture of intestinal contents in the antibiotic group and the antibiotic pancreatitis group were significantly different from those in the blank group, indicating that on the 7th day of the experiment, there was a disturbance of the flora and the proliferation of the fungi. 2. On the 14th day of the experiment, the positive results of fungal culture of intestinal contents in the antibiotic group and the antibiotic pancreatitis group were significantly different from those in the blank group, indicating that on the 14th day of the experiment, there was still a disturbance of the flora and the proliferation of the fungi. 3. There was no significant difference between the antibiotic group and the antibiotic pancreatitis group in the culture of intestinal contents fungi on the 7th and 14th day, indicating that the antibiotics were used for a long time. Load stress factors will not aggravate the disturbance of flora. 4. On the 7th day and 14th day, there was no evidence of breakthrough of intestinal barrier and no invasive fungal infection in the antibiotic group and the antibiotic pancreatitis group compared with the blank group. 5. The immune function of intestinal barrier plays an important role in the prevention of invasive mycosis. Conclusion: Long-term use of broad-spectrum antibiotics can cause intestinal microflora disorder and fungal proliferation. With the prolongation of antibiotic application time, load stress factors will not aggravate the flora disorder. The protective effect of intestinal inherent barrier can still ensure that under acute stress, fungi can not break through intestinal barrier, causing invasive mycosis, in which the immune function of intestinal barrier may play an important role.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R519

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