幼鼠急性肺损伤及CGRP干预下水通道蛋白1、5的表达水平研究
发布时间：2018-10-18 16:47
【摘要】：目的 了解降钙素基因相关肽(Calcitonin gene related peptide, CGRP)对脂多糖(lipopolysaccharide, LPS)诱导ALI动物模型干预过程中,水通道蛋白(Aquaporin, AQP)1和5的表达情况变化,探讨肺生理及病理条件下急性肺损伤与AQPs之间的关系；初步揭示CGRP对LPS诱导的ALI幼鼠水通道蛋白表达的调控作用。 方法 选用1月龄70-100g SD大鼠144只,按随机数字表法分为对照组(经腹腔注射等量生理盐水)、LPS组(腹腔注射LPS5mg/Kg)、CGRP干预组(LPS注射后10min,经腹腔注射CGRP1.0mg/Kg),每组48只,每个时间点每组8只。于注射LPS后0、2、6、12、24、48h处死。采用ELISA法测定LPS作用不同时间肺组织CGRP的分泌水平；光镜下观察肺损伤的严重程度；采用RT-PCR及Western blot法检测CGRP干预对于LPS诱导的ALI幼鼠AQP1、AQP5的表达变化。 结果 LPS处理后的大鼠肺组织匀浆中,CGRP的释放量不断增加,当在6h时,CGRP的表达量达到最高,与其他时间点相比差异具有显著性(p0.01),12h后,又迅速下降；AQP-1和AQP-5的mRNA表达水平迅速降低,均低于对照组,且差异具有显著性(p0.05),同时光镜下观察到肺水肿；CGRP干预后,AQP-1和AQP-5的mRNA表达水平增高,且相同时间点的表达量高于LPS组,但仍然低于对照组,差异均具有统计学意义(p0.05),同时光镜下观察到肺水肿较损伤组减轻。 结论 本实验选用大鼠作为实验对象建立脂多糖急性肺损伤动物模型,通过肺损伤相关指标的检测和病理组织学观察证明脂多糖致急性肺损伤动物模型建立成功,并观察AQP-1和AQP-5的mRNA及蛋白水平的表达变化及CGRP干预的影响,探讨急性肺损伤、AQPs、CGRP三者之间的联系,及CGRP的调控作用。结果提示如下： 1. CGRP可使急性肺损伤大鼠病理学改变减轻,说明CGRP对急性肺损伤有较好的保护作用。 2.AQP1和AQP5的表达量在急性肺损伤组较正常对照组显著下降,引起水肿液跨细胞膜的重吸收能力下降,进而导致急性肺损伤时肺水肿形成。 3. CGRP干预后,AQP-1和AQP-5的mRNA及蛋白表达水平均较急性肺损伤组显著增加,提示CGRP可上调LPS致ALI大鼠AQPs的表达,这是CGRP对ALI保护作用的机制之一。
[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.
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R725.6
本文编号：2279741
[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.
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R725.6
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相关期刊论文 前3条
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