To Explore The Mechanism Of Chemokines In Renal Tubular Injury In Diabetic Nephropathy Based On RNA-sequencing

Backgroud:The complex mechanism of kidney injury in Diabetic kidney disease(DKD)has not been fully elucidated.Studies have shown that inflammatory factors,including chemokines and cytokines in the renal microenvironment are closely related to kidney injury in DKD.A comprehensive and in-depth understanding of the expression,activity and molecular mechanism of chemokines related to renal injury in DKD will provide a reliable experimental basis for clinical treatment of renal injury in DKD.Abjective:To explore the expression and mechanism of chemokines in renal tubular injury induced by high glucose in diabetic nephropathy.Methods:⑴ The renal tubular cell injury model was established by glucose with different concents in vitro.⑵ The renal tubular injury cells were collected for RNA-Sequnecing.The differential expression in different groups were analyzed by using log2 Fold Change ≥1 and p <0.05 as the standard criteria.Differential gene expression profiles were obtaned and novel chemokines were screened.⑶ The expression and molecular mechanism of candidate novel chemokines in cell and clinical samples were detected and verified by using molecular biology techniques.⑷ The molecular mechanism of novel chemokines in high glucose induced renal injury were investigatied by using molecular biology techniques.Results:⑴ The renal tubular injury cell models were induced by different glucose concentrations for long-time.The cell activity and apoptosis were significantly decreased in the renal tubular injury cells.⑵ RNA sequencing combined with biosynthesis analysis was used to screen the novel chemokines.The CXCL2 and CCL20 were significantly overexpressed in renal tubular injury cell model induced by different glucose concentration for long-time.⑶ The leves of CXCL2 and CCL20 were significantly increased in clinical samples by using IHC and Q-PCR tests.⑷ The levels of CXCL2 and CCL20 were significantly increased in vitro by using Q-PCR and Western blot tests.The activity of MAPK kinase and Casepase 3 in apotosis related signaling pathway was significantly increased.Conclusin:⑴ The activity of renal tubular injury cells was significantly decreaed by different glucose concentrations induced long-time.And the Apoptosis was significantly increased by different glucose concentrations induced long-time.And the data proved there has a damaging effect on renal tubular cells.⑵ The most significant chemokines candidates were CXCL2 and CCL20 in the renal tubular injury model induced by different glucose concentrations based on RNA-seqencing.⑶ The levels of CXCL2 and CCL20 in renal tubular injury cells and diabetic nephropathy tissues were significantly higher than normal cells and chronic nephritis by IHC,Western blot and Q-PCR analysises.⑷ The new chemokines CXCL2 and CCL20 induced the renal tubular injury in diabetic nephropathy by promoting the activating apoptosis through MAPK pathway.