腺苷信号通路在大鼠单侧输尿管梗阻后肾纤维化作用机制研究

发布时间：2018-06-09 00:11

  本文选题：单侧输尿管梗阻 + 腺苷　； 参考：《中南大学》2014年硕士论文

【摘要】：目的：缺血和缺氧是导致慢性肾脏疾病的重要原因。腺苷作为缺血、缺氧性肾脏纤维化的重要信号分子,也可能是慢性肾脏疾病的一个重要致病因子。本研究检测了单侧输尿管梗阻后肾脏腺苷浓度的变化及其相关致纤维化因子的表达,探索了腺苷信号通路与肾纤维化的相关性,旨在进一步阐述慢性肾脏发病机制。 方法：选择清洁级健康雄性SD大鼠32只,随机分为两组：假手术组(对照组)和单侧输尿管梗阻组(UUO组),每组各16只,分别于手术后第1、2、3、4周分批处死前抽血,处死后取肾脏样本,每个时间点在两组中各取4只。分别用苏木素伊红(HE)、Masson染色观察各时间点大鼠肾组织病理变化以及肾间质胶原纤维沉积情况,应用图像分析系统进行半定量分析。采用高效液相色谱法(HPLC)检测肾组织腺苷水平,Elisa测定试剂盒检测血肌酐水平,实时定量PCR测定肾间质TGF-β1和Ⅰ型前胶原基因mRNA的表达量,免疫组织化学方法测定肾间质TGF-β1的蛋白表达,图像分析系统对免疫组化结果进行半定量分析。 结果：1、单侧输尿管梗阻后,实验组腺苷浓度相比对照组显著升高(P0.05),并且在实验后第一周达到高峰(P0.01),HE和Masson染色结果显示随着腺苷浓度显著增加,肾单位破坏逐渐增多,肾间质胶原沉积逐渐增加。2、在单侧输尿管梗阻后的四个时间点,未见血肌酐水平显著(P0.05)增加。3、实时PCR数据和免疫组化分析显示,在单侧输尿管梗阻后的四个时间点的TGF-β1和Ⅰ型前胶原的mRNA显著增加(P0.05),免疫组化分析表明在UUO后第二周,其在肾小管上皮细胞表达最多,然后随着时间推移,其表达逐渐下降,并且随着UUO时间的延长,TGF-β1的分布从肾小管上皮细胞向肾间质转移,进而蔓延至肾小球。 结论：1、肾间质纤维化过程中可能存在肾组织持续低氧,并导致组织内腺苷水平升高；2、腺苷含量增高,加速了肾小管损伤、肾间质纤维化的发生、发展；3、腺苷信号通路可能通过调节TGF-β1、 α1(Ⅰ)procollagen mRNA表达,干预RIF的发展。
[Abstract]:Objective: ischemia and hypoxia are important causes of chronic renal disease. Adenosine, as an important signal molecule of ischemia and hypoxic renal fibrosis, may also be an important pathogenic factor of chronic renal disease. In this study, the changes of adenosine concentration and the expression of fibrosis factors in the kidney after unilateral ureteral obstruction were detected, and the correlation between adenosine signaling pathway and renal fibrosis was explored. Methods: Thirty-two healthy male SD rats of clean grade were randomly divided into two groups: sham operation group (control group) and UUO group with unilateral ureteral obstruction (16 rats in each group). Blood samples were taken from the kidney at the 1st and 2nd week after the operation and 4 weeks after the operation. 4 rats in each group were collected at each time point. The pathological changes of renal tissue and the deposition of renal interstitial collagen fibers in rats were observed by Masson staining. The semi-quantitative analysis was carried out by image analysis system. High performance liquid chromatography (HPLC) was used to detect adenosine level in renal tissue and serum creatinine level was detected by Elisa kit. The expression of TGF- 尾 1 and procollagen 鈪，

本文编号：1997865