【摘要】：目的：探討谷氨酰胺對(duì)腸缺血再灌注損傷大鼠腸粘膜形態(tài)病理學(xué)的保護(hù)作用以及分子機(jī)制，并觀察不同的給藥方法、不同給藥時(shí)間對(duì)谷氨酰胺作用的影響。 方法：1.用手術(shù)的方法構(gòu)建大鼠腸缺血再灌注損傷模型。 2.酶聯(lián)免疫吸附試驗(yàn)（ELISA）檢測大鼠血標(biāo)本中TNF-α含量的變化。 3.腸標(biāo)本的石蠟包埋、切片、以及HE染色，觀察谷氨酰胺對(duì)大鼠腸缺血再灌注損傷形態(tài)學(xué)的影響。 4.用RT-PCR和Western-blot的方法檢測腸缺血再灌注損傷的各組大鼠腸標(biāo)本組織中NF-κB和PPAR-γ基因的mRNA和蛋白表達(dá)情況。 結(jié)果：1.腸缺血大鼠模型：正常喂養(yǎng)組（NG組）、給予尾靜脈注射谷氨酰胺組(GIG組)、給予谷氨酰胺灌胃組(GOG組)、術(shù)中腸系膜上靜脈給予谷氨酰胺組（GIOG組）、術(shù)中腸腔給予谷氨酰胺組（GIEC組）,模型構(gòu)建成功。 2.我們用ELISA的方法分別檢測五組大鼠(NG、GIG、GOG、GIOG、GIEC)血清中TNF-α的含量，我們發(fā)現(xiàn)1個(gè)小時(shí)、12個(gè)小時(shí)NG、GIEC組大鼠血清中TNF-α含量明顯高于GIG、GOG組（P＜0.05），同時(shí)也比GIOG組含量高。 3.我們用HE染色的方法，觀察了谷氨酰胺對(duì)大鼠腸粘膜缺血再灌注損傷形態(tài)學(xué)的影響。結(jié)果顯示NG、GIEC組大鼠在1小時(shí)和12小時(shí)腸粘膜的形態(tài)損傷非常嚴(yán)重。而GOG、GIG組大鼠1小時(shí)和12小時(shí)腸粘膜的損傷程度比NG、GIEC組輕。GIOG組大鼠1小時(shí)和12小時(shí)腸粘膜的損傷程度比GOG、GIG嚴(yán)重。 4.我們使用逆轉(zhuǎn)錄聚合酶鏈反應(yīng)技術(shù)（RT-PCR）和蛋白免疫印跡技術(shù)（Western-blot），去評(píng)估各組(NG、GIG、GOG、GIOG、GIEC)大鼠腸粘膜標(biāo)本中NF-κB和PPAR-γ基因的mRNA和蛋白表達(dá)情況。NG、GIEC組NF-κB和PPAR-γ基因的mRNA和蛋白表達(dá)量最高，兩組之間并沒有差異（P＞0.05）；GIG、GOG組NF-κB和PPAR-γ基因的mRNA和蛋白表達(dá)量最低，與NG、GIEC組NF-κB和PPAR-γ基因的mRNA和蛋白表達(dá)量有顯著的差異（P＜0.05）。 結(jié)論：1.對(duì)于腸缺血再灌注損傷的大鼠模型，提前4天預(yù)給予谷氨酰胺，，不管是靜脈注射組，還是口服組，谷氨酰胺都能夠?qū)Υ笫蟮哪c粘膜起到保護(hù)作用，能夠降低血清TNF-α水平；而在腸缺血時(shí)給藥，因?yàn)殪o脈注射使細(xì)胞內(nèi)很快能夠得到豐富的谷氨酰胺，所以能夠起到一定的保護(hù)作用，但是腸腔內(nèi)給藥，因?yàn)槿毖俟嘧p傷的存在并不能吸收，所以未對(duì)腸粘膜起到保護(hù)作用，也不能降低血清TNF-α的水平。 2、對(duì)于腸缺血再灌注損傷模型，谷氨酰胺通過降低NF-κB的活性和增加PPAR-γ活性的機(jī)制，對(duì)腸缺血再灌注損傷起到保護(hù)作用。
[Abstract]:Aim: to investigate the protective effect and molecular mechanism of glutamine on intestinal mucosal morphology in rats with intestinal ischemia-reperfusion injury, and to observe the effects of different administration methods and administration time on glutamine. Method 1: 1. The model of intestinal ischemia-reperfusion injury in rats was established by surgical method. 2. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of TNF- 偽 in blood samples of rats. The effects of glutamine on the morphology of intestinal ischemia-reperfusion injury in rats were observed by paraffin embedding, sectioning and HE staining. 4. The expression of mRNA and protein of NF- 魏 B and PPAR- 緯 genes in intestinal tissues of rats with intestinal ischemia-reperfusion injury was detected by RT-PCR and Western-blot. The result is 1: 1. Model of intestinal ischemia in rats: normal feeding group (NG group), caudal intravenous injection of glutamine group (GIG group), glutamine gastric perfusion group (GOG group), intraoperative superior mesenteric vein glutamine group (GIOG group), intraoperative lumen administration of glutamyl glutamate. Amines group (GIEC group), the model was successfully constructed. 2. The content of TNF- 偽 in serum of five groups of rats (NG,GIG,GOG,GIOG,GIEC) was detected by ELISA method. We found that the content of TNF- 偽 in serum of the NG,GIEC group was significantly higher than that of the GIG,GOG group (P < 0. 05) and was higher than that of the GIOG group after 1 hour and 12 hours of NG,GIEC (P < 0. 05), and was also higher than that of the GIOG group (P < 0. 05). The effects of glutamine on the morphology of intestinal mucosal ischemia reperfusion injury in rats were observed by HE staining. The results showed that the morphological damage of intestinal mucosa in NG,GIEC group was very serious at 1 h and 12 h. However, the degree of intestinal mucosal damage in GOG,GIG group was more severe than that in NG,GIEC group at 1 and 12 hours compared with that in NG,GIEC group. 4. We used reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot (Western-blot) to evaluate the expression of mRNA and protein of NF- 魏 B and PPAR- 緯 genes in the intestinal mucosa of rats in each group (NG,GIG,GOG,GIOG,GIEC). The mRNA and protein expression of NF- 魏 B and PPAR- 緯 genes were the highest in NGG IEC group. There was no difference between the two groups (P > 0. 05). The mRNA and protein expression of NF- 魏 B and PPAR- 緯 gene were the lowest in GIGGG group, and the mRNA and protein expression of NF- 魏 B and PPAR- 緯 gene in NG,GIEC group were significantly different (P < 0. 05). Conclusion 1. For the model of intestinal ischemia-reperfusion injury in rats, glutamine was given 4 days in advance. Both intravenous and oral groups, glutamine could protect intestinal mucosa and decrease serum TNF- 偽 level. And when given during intestinal ischemia, because intravenous injection can quickly make the cells rich in glutamine, so it can play a protective role, but intraluminal administration of drugs, because the presence of ischemia-reperfusion injury can not be absorbed, So it has no protective effect on intestinal mucosa, nor can it reduce the level of serum TNF- 偽. 2 for intestinal ischemia-reperfusion injury model, glutamine reduces the activity of NF- 魏 B and increases the activity of PPAR- 緯. It has protective effect on intestinal ischemia reperfusion injury.
【學(xué)位授予單位】：福建醫(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)】：R965

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