Abstract：
Chronic renal failure (CRF) is a severe syndrome affecting the urinary system for which there are no effective therapeutics. In this study, we investigate the effects and mechanisms of aminophylline in preventing CRF development. A rat model of chronic renal failure is established by 5/6 nephrectomy. The levels of serum creatinine (SCR), urinary protein (UPR), and blood urea nitrogen (BUN) are detected by ELISA. Histological evaluations of renal tissues are performed by H&E, Masson staining, and PAS staining. Functional protein expression is detected by western blot analysis or immunofluorescence microscopy. Glomerular cell apoptosis is determined using the TUNEL method. Results show that Aminophylline significantly reduces the levels of SCR, UPR, and BUN in the CRF model rats. Histological analyses show that aminophylline effectively alleviates renal tissue injuries in CRF rats. The protein expression levels of nephrin, podocin, SIRT1, p-AMPK, and p-ULK1 are greatly increased, while p-mTOR protein expression is markedly decreased by aminophylline treatment. Additionally, the protein level of LC3B in CRF rats is significantly increased by aminophylline. Moreover, aminophylline alleviates apoptosis in the glomerular tissues of CRF rats. Furthermore, resveratrol promotes SIRT1, p-AMPK, and p-ULK1 protein expressions and reduces p-mTOR and LC3B protein expressions in CRF rats. Selisistat (a SIRT1 inhibitor) mitigates the changes in SIRT1, p-AMPK, p-ULK1, p-mTOR, and LC3B expressions induced by aminophylline. Finally, RAPA alleviates renal injury and apoptosis in CRF rats, and 3-MA eliminates the aminophylline-induced inhibition of renal injury and apoptosis in CRF rats. Aminophylline suppresses chronic renal failure progression by modulating the SIRT1/AMPK/mTOR-mediated autophagy process.
摘要：
慢性肾衰竭（CRF）是一种严重的综合征，影响泌尿系统，没有有效的治疗方法。在这项研究中,我们研究了氨茶碱在预防CRF发展中的作用和机制。采用5/6肾切除术建立慢性肾功能衰竭大鼠模型。血清肌酐（SCR）水平，尿蛋白（UPR），通过ELISA检测血尿素氮(BUN)。肾组织的组织学评估由H&E，Masson染色,和PAS染色。通过蛋白质印迹分析或免疫荧光显微镜检测功能性蛋白质表达。使用TUNEL方法测定肾小球细胞凋亡。结果表明，氨茶碱显著降低SCR的水平，UPR,CRF模型大鼠的BUN。组织学分析表明，氨茶碱可有效减轻CRF大鼠的肾组织损伤。nephrin的蛋白质表达水平，波多辛,SIRT1,p-AMPK,和p-ULK1大大增加，而p-mTOR蛋白表达在氨茶碱处理下显著降低。此外,氨茶碱可显著提高CRF大鼠LC3B蛋白水平。此外，氨茶碱减轻CRF大鼠肾小球组织凋亡。此外，白藜芦醇促进SIRT1，p-AMPK，和p-ULK1蛋白的表达，并降低CRF大鼠的p-mTOR和LC3B蛋白的表达。Selissistat（SIRT1抑制剂）减轻SIRT1，p-AMPK，p-ULK1,p-mTOR,和氨茶碱诱导的LC3B表达。最后,RAPA减轻CRF大鼠肾损伤和细胞凋亡,3-MA消除了氨茶碱对CRF大鼠肾损伤和细胞凋亡的抑制作用。氨茶碱通过调节SIRT1/AMPK/mTOR介导的自噬过程抑制慢性肾衰竭进展.
