Submit Manuscript  

Article Details


The Mechanism of miR-192 in Regulating High Glucose-Induced MCP-1 Expression in Rat Glomerular Mesangial Cells

[ Vol. 19 , Issue. 7 ]

Author(s):

Fenqin Chen, Guozhu Wei, Yang Zhou, Xiaoyu Ma and Qiuyue Wang*   Pages 1055 - 1063 ( 9 )

Abstract:


Background: Although the pathogenetic mechanism of diabetic kidney disease (DKD) has not been elucidated, an inflammatory mechanism may be a potential contributor. Monocyte chemotactic protein-1(MCP-1) is suggested to be implicated in the development of DKD by playing a role in the infiltration of monocyte/macrophage. The aim of this study was to investigate the expression of MCP-1 under high glucose conditions, as well as the effects of microRNA-192 (miR-192) under these conditions, and to study the regulatory mechanism of MCP-1 in DKD.

Methods: Rat glomerular mesangial cells were cultured in high glucose or isotonic mannitol. The messenger RNA(mRNA) expression of miR-192, miR-200b, miR-200c, E-box-binding homeobox 1 (Zeb1), and MCP-1 was then detected by real-time PCR, and the protein expression of Zeb1 and MCP- 1 was assessed by western blotting. The rat mesangial cells were transfected with an miR-192 inhibitor, NC inhibitor , and transfected with siRNA Zeb1, siNC. The cells were then cultured in high glucose to detect the mRNA expression of miR-192, miR-200b, miR-200c, Zeb1, and MCP-1 using realtime PCR, and Zeb1 and MCP-1 protein expression were determined by western blotting.

Results: MiR-192, miR-200b, miR-200c, and MCP-1 were overexpressed, whereas Zeb1 was downregulated when cultured in high glucose (P < 0.05). After transfection with an miR-192 inhibitor, the expression of miR-192, miR-200b, miR-200c, and MCP-1 was downregulated, whereas Zeb1 was increased, and these differences were statistically significant (P < 0.05). The observed changes in the expression in the NC inhibitor transfection group were similar to that of non-transfected cell lines. Silencing the expression of Zeb1 resulted in a significant increase in the expression of miR-192, miR- 200b, miR-200c, and MCP-1 (P < 0.05). The observed changes in the SiNC transfection group were similar to those of non-transfected cell lines.

Conclusions: MiR-192 expression was upregulated to increase the expression of inflammatory factor MCP-1 by inhibiting the expression of Zeb1, which was mediated by breaking the regulatory loop of Zeb1 and miR-200b/c in rat mesangial cells cultured in high glucose.

Keywords:

High glucose, rat glomerular mesangial cells, miR-192, miR-200b, miR-200c, MCP-1.

Affiliation:

Departments of Geriatric, the First Affiliated Hospital, China Medical University, Shenyang 110001, Department of Radiology, Orthopedic Hospital of Shenyang, Shenyang 110001, Endocrinology, the First Affiliated Hospital, China Medical University, Shenyang 110001, Departments of Geriatric, the First Affiliated Hospital, China Medical University, Shenyang 110001, Endocrinology, the First Affiliated Hospital, China Medical University, Shenyang 110001

Graphical Abstract:



Read Full-Text article