晚期糖基化终产物受体系统在AOPP所致肾病中的作用及复方黄甘颗粒的干预研究

The accumulation of Plasma-Advanced Oxidation Protein Products (AOPP) is prevalent in chronic kidney disease (CKD). AOPP are a group of reversible and poisonous products formed by oxidant stress at the same time are potential circulating and tissue uremic toxins. Detoxification and pathogenic effects of AOPP are associated with advanced glycation endproducts (AGE)-receptor system. The express of AGE-receptor system are abnormal in CKD, which RAGE overexpress and AGE-R1 is suppressed. Whether AOPP is involved in AGE-receptor system expression and regulation in mesangial cells surface remain unclear. The prophase study was undertaken to determine that AOPP induce receptor for AGE (RAGE) expression and inhibitory AGE receptor 1 (AGE-R1) expression, which promote ROS generation. Traditional Chinese medication Huang Gan formula (HGF), which was the second generation of a famous traditional Chinese medication NiaoDuQing granule, was invented by NanFang Hospital. We have a proprietary right to the prescription. Potentially renal-protective mechanism of HGF is associated with AGE-receptor system..So we proposed a new hypothesis which is AOPP induce the abnormal expression of AGE-receptor system, cause ROS generation increasing and accelerate CKD depravation. The present study was undertaken to determine 1. The molecular mechanism of RAGE、AGE-R1 gene expression regulation by AOPP, which transcription factor will be involved in. 2. The signal transduction of expression of AGE-receptor system induced by AOPP in mesangial cells. 3. Huang Gan formula which might protect renal and other tissue injury due to metabolic inflammation in CKD, probably through regulate with AGE-receptor system on the cell surface. In addition, we investigate the therapeutic benefits of manipulation of AGE-receptor system expression and interactions as a therapeutic target for the treatment of CKD.

晚期氧化蛋白产物（AOPP）是慢性肾脏疾病（CKD）氧化应激的产物又可通过晚期糖基化终产物（AGE）受体系统产生病理作用。CKD患者AGE受体系统出现异常，RAEG表达上调而AGE-R1表达被抑制，AOPP是否以及如何参与该变化的发生还不清楚。我们前期研究表明：AOPP可诱导细胞RAGE表达，抑制AGE-R1表达，导致细胞活性氧（ROS）产生增加；尿毒清颗粒的二代产品，专利药复方黄甘颗粒可干预这一过程。在此基础上我们将深入研究AGE受体系统在AOPP致病和被清除中的作用1.何种正负性转录因子在AOPP影响系膜细胞RAGE、AGE-R1基因转录中起调控作用；2. AOPP通过何种信号转导通路导致AGE受体系统表达异常；3.复方黄甘颗粒调节AGE受体系统表达干预代谢性炎症的作用。本研究为通过调节AGE受体系统表达干预CKD进展的治疗新思路提供理论依据；对发现干预CKD的药物具有重要意义。

晚期氧化蛋白产物（AOPPs）蓄积是中国慢性肾脏病（CKD）患者代谢性紊乱的特征之一。AOPPs与晚期糖基化终产物受体（RAGE）结合，通过受体后信号产生病理生物学作用是导致代谢性紊乱的重要因素。而以AGE-R1为代表的受体与晚期糖基化终产物（AGE）降解、清除相关。我们在前期研究复方黄甘抗氧化应激与RAGE/AGE-R1的改变有关的基础上，通过研究复方黄甘颗粒延缓CKD进展的分子机制，阐明RAGE/AGE-R1系统作为CKD进展的干预靶标。复方黄甘可降低血清AOPPs和AGE水平，抑制RAGE mRNA 与蛋白表达，上调AGE-R1表达，并通过JAK2／STAT3途径调节AOPPs对AGE受体系统表达的平衡。主要成分芍药苷能阻断AOPPs与RAGE受体结合从而抑制Nox2／Nox4表达，减少ROS产生；抑制异常的Ca2+通量；恢复线粒体膜电位和ATP水平；抑制NF-κB及下游HIF-1的激活，减小氧化应激损伤。我们还研究了AOPPs影响肝脏ABC类转运体表达的，从而影响体内物质的代谢。用荧光素酶报告基因质粒，通路抑制剂ML385，PDTC，BAY117082，siRNA的方法检测AOPPs对Nrf-2，NF-κB，Foxo3α等通路的作用。AOPPs降低HDAC2的mRNA和蛋白表达；增加调控因子Foxo3α及通路相关蛋白表达及核转移；增强ABCB1的转运活性；降低组蛋白乙酰化酶27位的甲基化，增加组蛋白乙酰化酶4位的甲基化。本项目通过研究复方黄甘调节CKD时RAGE、AGE-R1受体的异常表达，从而延缓CKD的进展，为中医药用于治疗CKD提供理论依据。阐明了AOPPs上调RAGE，下调AGE-R1的表达机制，为防治CKD进展药物研发提供新靶点。