【摘要】：目的 了解降鈣素基因相關(guān)肽(Calcitonin gene related peptide, CGRP)對(duì)脂多糖(lipopolysaccharide, LPS)誘導(dǎo)ALI動(dòng)物模型干預(yù)過(guò)程中,水通道蛋白(Aquaporin, AQP)1和5的表達(dá)情況變化,探討肺生理及病理?xiàng)l件下急性肺損傷與AQPs之間的關(guān)系；初步揭示CGRP對(duì)LPS誘導(dǎo)的ALI幼鼠水通道蛋白表達(dá)的調(diào)控作用。 方法 選用1月齡70-100g SD大鼠144只,按隨機(jī)數(shù)字表法分為對(duì)照組(經(jīng)腹腔注射等量生理鹽水)、LPS組(腹腔注射LPS5mg/Kg)、CGRP干預(yù)組(LPS注射后10min,經(jīng)腹腔注射CGRP1.0mg/Kg),每組48只,每個(gè)時(shí)間點(diǎn)每組8只。于注射LPS后0、2、6、12、24、48h處死。采用ELISA法測(cè)定LPS作用不同時(shí)間肺組織CGRP的分泌水平；光鏡下觀察肺損傷的嚴(yán)重程度；采用RT-PCR及Western blot法檢測(cè)CGRP干預(yù)對(duì)于LPS誘導(dǎo)的ALI幼鼠AQP1、AQP5的表達(dá)變化。 結(jié)果 LPS處理后的大鼠肺組織勻漿中,CGRP的釋放量不斷增加,當(dāng)在6h時(shí),CGRP的表達(dá)量達(dá)到最高,與其他時(shí)間點(diǎn)相比差異具有顯著性(p0.01),12h后,又迅速下降；AQP-1和AQP-5的mRNA表達(dá)水平迅速降低,均低于對(duì)照組,且差異具有顯著性(p0.05),同時(shí)光鏡下觀察到肺水腫；CGRP干預(yù)后,AQP-1和AQP-5的mRNA表達(dá)水平增高,且相同時(shí)間點(diǎn)的表達(dá)量高于LPS組,但仍然低于對(duì)照組,差異均具有統(tǒng)計(jì)學(xué)意義(p0.05),同時(shí)光鏡下觀察到肺水腫較損傷組減輕。 結(jié)論 本實(shí)驗(yàn)選用大鼠作為實(shí)驗(yàn)對(duì)象建立脂多糖急性肺損傷動(dòng)物模型,通過(guò)肺損傷相關(guān)指標(biāo)的檢測(cè)和病理組織學(xué)觀察證明脂多糖致急性肺損傷動(dòng)物模型建立成功,并觀察AQP-1和AQP-5的mRNA及蛋白水平的表達(dá)變化及CGRP干預(yù)的影響,探討急性肺損傷、AQPs、CGRP三者之間的聯(lián)系,及CGRP的調(diào)控作用。結(jié)果提示如下： 1. CGRP可使急性肺損傷大鼠病理學(xué)改變減輕,說(shuō)明CGRP對(duì)急性肺損傷有較好的保護(hù)作用。 2.AQP1和AQP5的表達(dá)量在急性肺損傷組較正常對(duì)照組顯著下降,引起水腫液跨細(xì)胞膜的重吸收能力下降,進(jìn)而導(dǎo)致急性肺損傷時(shí)肺水腫形成。 3. CGRP干預(yù)后,AQP-1和AQP-5的mRNA及蛋白表達(dá)水平均較急性肺損傷組顯著增加,提示CGRP可上調(diào)LPS致ALI大鼠AQPs的表達(dá),這是CGRP對(duì)ALI保護(hù)作用的機(jī)制之一。
[Abstract]:Objective to investigate the changes of the expression of aquaporin (Aquaporin, AQP) 1 and 5 during the intervention of calcitonin gene-related peptide (Calcitonin gene related peptide, CGRP) on ALI model induced by lipopolysaccharide (lipopolysaccharide, LPS). To explore the relationship between acute lung injury and AQPs under physiological and pathological conditions of lung, and to reveal the regulatory effect of CGRP on the expression of aquaporin in ALI young rats induced by LPS. Methods 144 1-month-old 70-100g SD rats were randomly divided into control group (48 rats in each group) in which 48 rats in each group were treated by intraperitoneal injection of normal saline), LPS (intraperitoneal injection of LPS5mg/Kg), CGRP for 10 min and intraperitoneal injection of CGRP1.0mg/Kg for 10 min). There were 8 rats in each group at each time point. The rats were killed at 24 hours after injection of LPS. The levels of CGRP secretion in lung tissue were measured by ELISA method, the severity of lung injury was observed under light microscope, and the expression of AQP1,AQP5 in LPS induced ALI young rats was detected by RT-PCR and Western blot. Results the release of CGRP in the homogenate of lung tissue of rats treated with LPS increased continuously, and the expression of CGRP reached the highest level at 6h, which was significantly different from that at other time points (p0.01), and then decreased rapidly after 12h. The expression of mRNA in AQP-1 and AQP-5 decreased rapidly, which was significantly lower than that in the control group (p0.05), and pulmonary edema was observed under light microscope, the mRNA expression of AQP-1 and AQP-5 increased after CGRP intervention, and the expression of mRNA was higher than that of LPS group at the same time. But it was still lower than the control group, the difference was statistically significant (p0.05), at the same time, the pulmonary edema was lighter than that in the injured group under light microscope. Conclusion the animal model of acute lung injury induced by lipopolysaccharide was established in rats. The animal model of acute lung injury induced by lipopolysaccharide was successfully established by the detection of relevant indexes of lung injury and histopathological observation. The expression of mRNA and protein in AQP-1 and AQP-5 and the effect of CGRP intervention were observed to explore the relationship between AQPs,CGRP and acute lung injury and the regulation of CGRP. The results are as follows: 1. The pathological changes of acute lung injury rats were alleviated by CGRP, which indicated that CGRP had a better protective effect on acute lung injury. The expression of 2.AQP1 and AQP5 in the acute lung injury group was significantly lower than that in the normal control group. The reabsorption ability of edema fluid across cell membrane was decreased, and pulmonary edema was formed in acute lung injury. 3. 3%. After CGRP intervention, the expression of mRNA and protein in AQP-1 and AQP-5 were significantly higher than those in acute lung injury group, suggesting that CGRP can up-regulate AQPs expression in ALI rats induced by LPS, which is one of the mechanisms of CGRP's protective effect on ALI.
【學(xué)位授予單位】：大理學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R725.6

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本文編號(hào)：2279741