miR-30s靶向调控介导糖尿病肾脏疾病足细胞损伤及固本化瘀通络方对其干预作用及机制研究

It has been proved that DKD(diabetic kidney disease) is caused by podocyte injury during early stage of development, but the mechanism of this pathological change has not been known clearly. Our previous study have found that there are several abnormal expressed microRNAs in kidney tissue of DKD patients, which probably attend the development of disease. So,as an entry point to AGEs-RAGE in podocyte injury, we proposed that miR-30a probably regulate AGEs-RAGE signaling pathway by targeting RAGE gene in the process of DKD occurrence according to previous literature and experimental results. In order to prove above hypothesis, we’ll carry out this experiment in three aspects. First,we’ll detect abnormal expressed miR-30s in kidney of DKD patients. Second,to detect the expression level of miR-30s, AGEs,RAGE,nephrin and podocin of podocyte in high glucose environment and exam the regulation of miR-30a in this process. Finally,we’ll focus on the effect and mechanism of Guben Huayu Decoction in podocyte injury caused by high glucose both in vivo and vitro experiments. On the basis of experiments above, we conclude that miR-30a participate in the occurrence and development of DKD and Guben Huayu Decoction protect podocyte by targeting on miR-30a theoretically. This study revealed the mechanism of Traditional Chinese Medicine on the point of miRNA. This provides the cornerstone of modernization of Traditional Chinese Medicine.

糖尿病肾脏疾病（DKD）是一种足细胞疾病已成为一种共识，足细胞损伤在DKD的早期就已经发生，而对于足细胞损伤的发生机制目前研究并不十分清楚。既往我们研究发现，DKD患者肾脏组织存在多种异常表达的microRNA，这些异常表达的miRNA可能参与了DKD的发生和发展。因此，本研究立足于足细胞，以AGEs-RAGE对足细胞的损伤为切入点，着眼于体内miRNA的作用，结合文献及前期实验结果，提出miR-30a可能通过靶向调节RAGE基因的表达从而调控AGEs-RAGE信号通路，参与DKD足细胞损伤过程。主要通过以下几个方面进行研究：①DKD患者肾脏组织miR-30s的异常表达情况；②高糖环境下足细胞miR-30s、AGEs、RAGE、nephrin、podocin等的表达变化及miR-30a的调控作用；③固本化瘀通络方对于高糖所致足细胞损伤（体内及体外实验）的保护作用及机制。

糖尿病肾脏疾病（DKD）是一种足细胞疾病已成为一种共识，足细胞损伤在DKD的早期就已经发生，而对于足细胞损伤的发生机制目前研究并不十分清楚。既往我们研究发现，DKD患者肾脏组织存在多种异常表达的microRNA，这些异常表达的miRNA可能参与了DKD的发生和发展。因此，本研究立足于足细胞，以AGEs-RAGE对足细胞的损伤为切入点，着眼于体内miRNA的作用，结合文献及前期实验结果，提出miR-30a通过靶向调节SOCS1基因的表达影响JAK/STAT信号通路从而调控AGEs-RAGE的表达，参与DKD足细胞损伤过程。研究结果主要有以下几个方面：①DKD患者肾脏组织中miR-30a表达明显增加；②体外高糖环境下足细胞miR-30a、AGEs、RAGE、nephrin、podocin、JAK1/JAK2、STAT1/STAT2表达均上调，而SOCS1基因表达下调，使用miR-30a抑制慢病毒感染足细胞后发现，miR-30a、AGEs、RAGE、nephrin、podocin、JAK1/JAK2、STAT1/STAT2 表达均明显下降，而SOCS1基因表达上调；③荧光素酶报告基因证实SCOS1为miR-30a靶向调控基因。④体外高糖环境下的足细胞，在加入固本化瘀通络方干预后，可抑制miR-30a、AGEs、RAGE、nephrin、podocin、JAK1/JAK2、STAT1/STAT2的表达，伴随nephrin、podocin的降低；⑤糖尿病SD大鼠肾脏miR-30a表达明显增加，伴随AGEs、RAGE、nephrin、podocin、JAK1/JAK2、STAT1/STAT2表达均上调，与体外实验结果一致，而体内给予中药固本化瘀通络方可下调上述基因的表达，降低蛋白尿，减轻DKD肾脏损伤。通过上述研究，我们提出，miR-30a参与DKD的发生发展，中药固本化瘀通络方通过miR-30a发挥了DKD足细胞保护作用，进一步从miRNA层面揭示中药的作用机理，为中医药的现代化发展奠定基础。