P53在急性肾损伤小鼠肾脏的表达及其与细胞凋亡关系的研究

发布时间：2018-02-24 23:14

本文关键词： p53 急性肾损伤 PFT-α 凋亡缺血再灌注　出处：《南京大学》2011年硕士论文　论文类型：学位论文

【摘要】：实验一P53在缺血再灌注导致急性肾损伤小鼠肾脏的表达目的：探讨缺血再灌注致急性肾损伤(AKI)小鼠肾脏不同部位p53的表达及细胞凋亡的变化。方法：采用随机对照动物实验方法,将12只小鼠随机分为两组：假手术组和AKI组,每组6只。采用双侧肾蒂夹闭45 min后松开动脉夹的方法建立小鼠AKI模型,于建模后48 h采血检测尿素氮、肌酐,取肾脏组织行HE染色观察肾组织病理学变化,采用Western Blot法测定p53蛋白含量,免疫荧光法确定肾脏不同部位p53的表达,TUNEL法检测肾脏细胞凋亡,免疫组化方法检测TNFR、Caspase-3及Bcl-2蛋白水平。结果：(1)AKI组小鼠血尿素氮和肌酐水平均明显高于假手术组(P0.01)；HE染色显示假手术组肾组织细胞形态完整,排列整齐,无明显病理改变,AKI组则表现为肾小管上皮细胞刷状缘脱落、空泡及滴状变性,皮髓质间有明显淤血带；(2)p53在假手术组小鼠肾脏无表达,而AKI组小鼠缺血再灌注48h后p53蛋白则明显增加(P0.05),并且主要在肾皮质表达；(3)假手术组小鼠肾脏未检测到凋亡细胞,AKI组小鼠肾脏凋亡细胞指数明显增加(P0.01)；TNFR蛋白及Caspase-3蛋白水平升高(P0.01),Bcl-2蛋白水平下降(P0.01)。结论：急性肾损伤时,肾组织p53表达增加,主要定位于皮质,并可能参与介导细胞凋亡实验二急性肾损伤小鼠肾脏P53表达与细胞凋亡的关系目的：探讨缺血再灌注致AKI小鼠肾脏p53的表达与细胞凋亡的关系。方法：采用随机对照动物实验方法,将18只小鼠随机分为三组：假手术组、AKI组及PFT-α组,每组6只。采用双侧肾蒂夹闭45 min后松开动脉夹的方法建立小鼠AKI模型,PFT-α组于建模前5 min腹腔注射p53抑制剂(PFT-α)2.2mg/kg,并于建模后48 h采血检测尿素氮、肌酐,取肾脏组织行HE染色观察肾组织病理学变化,采用Western Blot法测定p53蛋白含量,TUNEL法检测肾脏细胞凋亡免疫组化方法检测(?)TNFR、Caspase-3及Bcl-2蛋白水平。结果：(1)PFT-α组和AKI组小鼠血尿素氮、肌酐水平均明显高于假手术组,而PFT-α组与AKI组比较血尿素氮、肌酐水平明显降低(P0.01)；肾组织HE染色显示AKI组肾小管上皮细胞刷状缘脱落、空泡及滴状变性,皮髓质间有明显淤血带；PFT-α组肾小管上皮部分刷状缘脱落消失,空泡及滴状变性减轻,皮髓质间无明显淤血带；(2)与AKI组相比,PFT-α组小鼠肾脏p53蛋白含量减少(P0.01),凋亡细胞指数也明显减少(P0.01); TNFR蛋白及Caspase-3蛋白水平也明显降低(P0.01), Bcl-2蛋白水平升高(P0.01)。结论：急性肾损伤时,p53可能通过调控凋亡蛋白Bcl-2.、TNFR蛋白水平,促进Caspase-3的释放,从而介导细胞凋亡。
[Abstract]:Expression of p53 in Kidney of mice with Acute Renal injury induced by Ischemia and reperfusion. Objective: to investigate the expression of p53 and apoptosis in different parts of kidney of acute renal injury (AKI) mice induced by ischemia-reperfusion. Methods: twelve mice were randomly divided into two groups: sham operation group and AKI group. The AKI model of mice was established by bilateral renal pedicle occlusion for 45 min and release of artery clamp. Blood samples were collected 48 hours after modeling to detect urea nitrogen and creatinine, and kidney tissues were stained with HE to observe the pathological changes of renal tissue. Western Blot method was used to detect p53 protein content, and immunofluorescence assay was used to determine the expression of p53 in different parts of kidney. Apoptosis of renal cells was detected by Tunel method. Results the levels of blood urea nitrogen and creatinine in the control group were significantly higher than those in the sham operation group. In the AKI group, the brush edge of renal tubular epithelial cells was shedding, vacuolation and degenerative, and there was no expression of p53 in the kidney of sham-operated mice. In AKI group, p53 protein increased significantly 48 hours after ischemia and reperfusion, and was mainly expressed in renal cortex. No apoptotic cells were detected in kidney of AKI group. The apoptotic cell index of kidney in AKI group was significantly higher than that in control group. Both P0.01 TNFR protein and Caspase-3 egg were significantly increased in AKI group. The level of Bcl-2 protein decreased with the increase of white level. Conclusion: during acute renal injury, the expression of Bcl 2 protein is decreased in patients with acute renal injury. The expression of p53 in renal tissue is increased, mainly located in the cortex, and may be involved in mediating apoptosis. The relationship between the expression of p53 and apoptosis in the Kidney of mice with Acute Renal injury. Aim: to investigate the relationship between the expression of p53 and apoptosis in the kidney of AKI mice induced by ischemia reperfusion. Methods: eighteen mice were randomly divided into three groups: sham-operated group and PFT- 伪 group. Six rats in each group were treated with bilateral renal pedicle occlusion for 45 min and then loosened the artery clamp. The AKI model was established by intraperitoneal injection of p53 inhibitor PFT- 伪 2.2 mg / kg at 5 min before modeling. Blood urea nitrogen and creatinine were measured 48 h after modeling. The histopathological changes of renal tissues were observed by HE staining. The content of p53 protein was determined by Western Blot method and the apoptosis of renal cells was detected by Tunel method. Results the levels of blood urea nitrogen and creatinine in the two groups were significantly higher than those in the sham operation group, and the blood urea nitrogen levels in the PFT- 伪 group and the AKI group were significantly higher than those in the AKI group, the levels of Caspase-3 and Bcl-2 protein in the TNFR- 伪 group were significantly higher than those in the sham operation group. The renal tissue HE staining showed that the brush edge, vacuole and degeneration of renal tubule epithelium in AKI group were obviously decreased, and that in PFT- 伪 group, the tubule epithelium brushed edge disappeared, vacuolation and droplet degeneration were alleviated in PFT- 伪 group. Compared with AKI group, the content of p53 protein in kidney of PFT- 伪 group decreased significantly, the apoptotic cell index also decreased significantly, the levels of TNFR protein and Caspase-3 protein decreased significantly, and the level of Bcl-2 protein increased P0.01a. Conclusion: P0.01protein level of PFT- 伪 group is higher than that of PFT- 伪 group, and the apoptotic cell index of P0.01a group is significantly lower than that of PFT- 伪 group, and the level of TNFR protein and Caspase-3 protein is also significantly lower than that of AKI group. During renal injury, p53 may regulate the level of apoptotic protein Bcl-2. Promote the release of Caspase-3, thus mediating cell apoptosis.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

【相似文献】