天然反义LncRNA-ARAP1-AS1和ARAP1-AS2协同调控ARAP1在糖尿病肾病发病机制和诊断中的作用

在前期实验基础上，验证糖尿病肾病（DN)患者血清、尿液和肾组织、体外培养的足细胞、系膜细胞、内皮细胞和肾小管上皮细胞、2型糖尿病db/db小鼠肾脏天然反义lncRNA-ARAP1-AS1、ARAP1-AS2及其正义链基因ARAP1的表达变化，探讨ARAP1-AS1和ARAP1-AS2对ARAP1的协同调控机制。明确ARAP1-AS1和ARAP1-AS2能否通过调控糖尿病状态下ARAP1的过表达，激活下游信号通路Rho/Cdc42和EGFR介导的PI3K/Akt/mTOR、ERK1/2、TGF-β1/Smad2/3、AMPK通路，促进肾脏固有细胞骨架重排、自噬抑制、凋亡、EMT、氧化应激、内质网应激、炎症和纤维化，探讨靶向抑制ARAP1-AS1和ARAP1-AS2异常表达能否减轻糖尿病肾脏损伤。验证结合血清、尿液ARAP1-AS1和ARAP1-AS2的诊断标签能否成为DN无创性生物标志物

Our previous studies found the expression of serum lncRNA-ARAP1-AS1 were gradually downregulated, lncRNA-ARAP1-AS2 was gradually upregulated and their sense strand gene ARAP1((ArfGAP with RhoGAP domain, ankyrin repeat and PH domain 1) mRNA was gradually upregulated along with the progression of diabetes and diabetic nephropathy. The diagnosis signature combined with the serum expression levels of lncRNA-ARAP1-AS1 and ARAP1-AS2 present a good diagnostic efficacy in diabetes mellitus and diabetic nephropathy. In present study, the expression levels of lncRNA-ARAP1-AS1，ARAP1-AS2 and ARAP1 in serum, urine and kidney tissue of diabetic nephropathy patients, high glucose cultured podocytes, mesangial cells, endothelial cells and renal tubular epithelial cells, and type 2 diabetes db/db mice kidneys will be detected. The coordinately regulatory mechanism of ARAP1-AS1 and ARAP1-AS2 on ARAP1 will be investigated. The objective is to elucidate the role of ARAP1-AS1 and ARAP1-AS2 on Rho/Cdc42, EGFR and its downstream pathway including PI3K/Akt/mTOR, ERK1/2, TGFβ1/Smad2/3, AMPK by upregulating ARAP1 to promote renal innate cells cytoskeleton rearrangement, autophagy inhibition, apoptosis, EMT, oxidative stress, endoplasmic reticulum stress, inflammation and fibrosis in diabetic nephropathy. The protective role of abated renal abnormal ARAP1-AS1, ARAP1-AS2 and ARAP1 expression on diabetic kidney injury will be observed. Furthermore, the value of diagnosis signature combined with the serum and urine lncRNA-ARAP1-AS1 and ARAP1-AS2 expression as biomarkers for diabetic kidney injury will be evaluated.