本文選題：化療 + 5-FU��； 參考：《第三軍醫(yī)大學(xué)》2012年碩士論文

【摘要】：目的： 本研究旨在探索腸粘膜屏障的保護(hù)方法。腸粘膜屏障是機體內(nèi)外聯(lián)系的重要界面，包括化療在內(nèi)的多種應(yīng)激因素可致腸粘膜屏障損壞�；熕幬镌跉缒[瘤細(xì)胞的同時，對消化道粘膜上皮組織細(xì)胞產(chǎn)生毒性。導(dǎo)致腸粘膜萎縮、絨毛變短腸通透性增加、腸局部免疫功能受損以及腸菌群失調(diào)等�；煂δc粘膜屏障的這些損傷可導(dǎo)致患者惡心、嘔吐、腹瀉，嚴(yán)重則會出現(xiàn)粘膜壞死、脫落、便血及全身不良反應(yīng)，甚至導(dǎo)致病情惡化，危及生命。因此，在化療時保護(hù)腸粘膜屏障結(jié)構(gòu)完整和功能正常是防治化療不良反應(yīng)的重要課題。 氨基葡萄糖(glucosamine, GlcN)是在2位碳上氨基取代的葡萄糖，N-乙酰氨基葡萄糖(N-Acetyl-D-glucosamine, GlcNAc)則是GlcN的氨基上發(fā)生乙�；难苌�，分子量小，水溶性好，具有抗氧化、免疫調(diào)節(jié)作用；能有效改善腸粘膜局部生態(tài)環(huán)境、提高上皮細(xì)胞活性、能進(jìn)入細(xì)胞發(fā)揮蛋白的糖基化修飾等作用，為此，本研究觀察GlcNAc對5-FU引起的腸粘膜結(jié)構(gòu)與功能損傷的保護(hù)作用，并探索其作用機理，為化療期間腸粘膜屏障受損新的防治方法研究奠定實驗基礎(chǔ)。 材料與方法： 1．動物的分組與模型建立：SPF級SD雄性大鼠50只，體重190~210g，隨機分為空白對照組、5-FU模型組、Gln組、GlcN組和GlcNAc組(n=10)。采用5-FU腹腔注射方法制備大鼠腸粘膜屏障損傷模型，按分組分別給藥，連續(xù)6d； 2．實驗過程中，密切檢測大鼠的體重變化；所有實驗動物每天稱量體重，并以此為依據(jù)調(diào)整給藥劑量； 3．采用HE染色法，，光鏡下觀察大鼠小腸粘膜組織病理形態(tài)學(xué)改變；通過透射電鏡，觀察小腸粘膜上皮細(xì)胞和細(xì)胞連接超微結(jié)構(gòu)改變； 4．采用分光光度法檢測大鼠血漿中D-乳酸的含量；采用ELISA法檢測大鼠血漿中DAO的含量； 5．Western blot法檢測大鼠小腸組織中O-GlcNAc蛋白的表達(dá)。 結(jié)果： 1.GlcNAc對大鼠體重變化的影響及解剖時大鼠小腸外觀觀察: 與實驗前相比，空白對照組、Gln組和GlcNAc組大鼠體重增加，而5-FU模型組和GlcN組大鼠體重減輕。解剖可見，空白對照組和GlcNAc組大鼠小腸無明顯變化，5-FU組、GlcN和Gln組不同程度存在腸脹氣、腸粘膜充血以及腹瀉。 2.GlcNAc對大鼠小腸組織病理形態(tài)學(xué)改變的影響: HE染色高倍鏡觀察顯示：空白對照組大鼠小腸腸壁結(jié)構(gòu)清晰完整，粘膜絨毛較細(xì)長，發(fā)達(dá)，排列規(guī)則緊密；5-FU模型組和GlcN組損傷明顯；GlcNAc組和Gln組大鼠小腸粘膜有輕度損傷表現(xiàn)。 3.GlcNAc對大鼠小腸組織超微結(jié)構(gòu)變化的影響 透射電鏡觀察顯示：5-FU細(xì)胞連接結(jié)構(gòu)局部出現(xiàn)異常；GlcNAc組大鼠小腸上皮細(xì)胞緊密連接結(jié)構(gòu)與空白對照組相似。 4.GlcNAc對大鼠血漿中D-乳酸含量的影響 GlcNAc組大鼠D-乳酸的含量較5-FU模型組和GlcN組顯著降低(p0.01)，分別降低了31.5%和33.7%。而GlcN組與5-FU模型組相比較沒有統(tǒng)計學(xué)差異(p0.05)。 5.GlcNAc對大鼠血漿中DAO含量的影響 與模型組和GlcN組相比較，Gln組和GlcNAc組大鼠DAO的含量明顯降低(p0.01)；而GlcN組與5-FU模型組相比較沒有統(tǒng)計學(xué)差異(p0.05)。 6.GlcNAc對大鼠小腸組織中O-GlcNAc蛋白表達(dá)的影響 與空白對照組相比，5-FU模型組和GlcN組大鼠小腸組織中O-G1cNAc蛋白水平明顯降低(p0.01)；與模型組和GlcN組相比較，Gln組和GlcNAc組大鼠O-G1cNAc蛋白水平明顯增高(p0.01)；而GlcN組與5-FU模型組相比較沒有統(tǒng)計學(xué)差異(p0.05)。 結(jié)論： 1．N-乙酰氨基葡萄糖能減輕化療所致大鼠腸粘膜損傷，降低腸粘膜通透性，保護(hù)小腸粘膜及其屏障功能； 2． N-乙酰氨基葡萄糖保護(hù)小腸粘膜及其屏障功能可能與上調(diào)小腸組織中O-GlcNAc蛋白表達(dá)有關(guān)。
[Abstract]:Objective:
The purpose of this study is to explore the protection of intestinal mucosal barrier. The intestinal mucosal barrier is an important interface between the internal and external contact of the body, and a variety of stress factors, including chemotherapy, can cause damage to the intestinal mucosal barrier. The chemotherapeutic drugs are toxic to the epithelial tissue cells of the digestive tract mucosa while killing the tumor cells. The intestinal mucosa atrophy and the villi become shorter. Intestinal mucosal permeability increases, intestinal immune function is damaged, and intestinal flora imbalance. These injuries to the intestinal mucosal barrier can cause nausea, vomiting, diarrhea, and severe mucosal necrosis, exfoliation, blood and systemic adverse reactions, even exacerbation of life. Therefore, protective intestinal mucosal barrier structure is protected during chemotherapy. The normal function of integrin is an important topic in the prevention and treatment of adverse reactions of chemotherapy.
Glucosamine (glucosamine, GlcN) is an amino - substituted glucose in 2 bits of carbon, and N- acetyl glucosamine (N-Acetyl-D-glucosamine, GlcNAc) is a derivative of acetylation on the amino group of GlcN, with small molecular weight, good water solubility, antioxidant and immune modulation; it can effectively improve the local ecological environment of the intestinal mucosa and improve the epithelium. In this study, the protective effect of GlcNAc on the structural and functional damage of intestinal mucosa caused by 5-FU was observed, and the mechanism of its action was explored, which lay the foundation for the study of new methods for preventing and controlling intestinal mucosal barrier during chemotherapy.
Materials and methods:
1. group and model of animals: 50 SPF male SD male rats, weight 190~210g, were randomly divided into blank control group, 5-FU model group, Gln group, GlcN group and GlcNAc group (n=10). The intestinal mucosal barrier damage model of rats was prepared by 5-FU intraperitoneal injection.
2. during the experiment, the weight changes of rats were closely monitored; all the experimental animals weighed daily and adjusted the dosage according to this.
3. HE staining was used to observe the pathological changes of the intestinal mucosa of the rats under light microscope, and the ultrastructural changes of the epithelial cells and cells of the small intestinal mucosa were observed by transmission electron microscopy.
4. the content of D- lactate in rat plasma was detected by spectrophotometry and the content of DAO in plasma was detected by ELISA.
5.Western blot was used to detect the expression of O-GlcNAc protein in the small intestine of rats.
Result:
Effects of 1.GlcNAc on body weight and the appearance of small intestine in rats during dissection:
Compared with before the experiment, the weight of the blank control group, the Gln group and the GlcNAc group increased, while the weight of the 5-FU model group and the group GlcN decreased, and the small intestine of the blank control group and the GlcNAc group had no obvious changes. In group 5-FU, there were flatulence, intestinal mucous membrane congestion and diarrhea in the GlcN and Gln groups.
Effects of 2.GlcNAc on the pathomorphological changes of small intestine in rats:
The observation of HE staining with high magnification showed that the intestinal wall structure of the small intestine in the blank control group was clear and complete, the mucous villi were long, developed, and the arrangement of the intestinal mucosa was close, the 5-FU model group and the GlcN group were injured obviously, and the small intestinal mucosa of the rats of group GlcNAc and Gln group had mild damage.
Effect of 3.GlcNAc on ultrastructural changes of small intestine in rats
Transmission electron microscopy showed that 5-FU cells were abnormal in connection structure. The tight junction structure of intestinal epithelial cells in GlcNAc group was similar to that in blank control group.
Effect of 4.GlcNAc on the content of D- lactate in rat plasma
The content of D- lactate in group GlcNAc rats was significantly lower than that in 5-FU model group and GlcN group (P0.01), which decreased by 31.5% and 33.7%. respectively, while there was no statistical difference between the GlcN group and the 5-FU model group (P0.05).
The effect of 5.GlcNAc on the content of DAO in rat plasma
Compared with the model group and the GlcN group, the content of DAO in the Gln group and the GlcNAc group decreased significantly (P0.01), but there was no statistical difference between the GlcN group and the 5-FU model group (P0.05).
Effect of 6.GlcNAc on O-GlcNAc protein expression in rat small intestine
Compared with the blank control group, the O-G1cNAc protein level in the small intestine of the 5-FU model group and the GlcN group was significantly lower (P0.01). Compared with the model group and the GlcN group, the level of O-G1cNAc protein in the Gln and GlcNAc groups increased significantly (P0.01), but there was no statistical difference between the GlcN group and the 5-FU model group (P0.05).
Conclusion:
1.N- acetylglucosamine can alleviate intestinal mucosal injury induced by chemotherapy, reduce intestinal permeability and protect intestinal mucosa and barrier function.
The protective effect of 2.N- acetylglucosamine on intestinal mucosal and barrier function may be related to the up regulation of O-GlcNAc protein in the small intestine.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R363

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