High glucose stimulates cell proliferation and Collagen IV production in rat mesangial cells through inhibiting AMPK-KATP signaling

Abstract

Purpose: The present study investigated the putative mechanisms underlying effects of K_ATP channel on high glucose (HG)-induced mesangial cell proliferation and tissue inhibitors of metalloproteinases (TIMP)-2 and Collagen IV production.

Methods: Rat mesangial cells were subjected to whole cell patch clamp to record the K_ATP channel currents under high glucose (HG, 30 mM) condition. Cell proliferation was measured using a CCK-8 assay. The production of TIMP-2 and Collagen IV and AMP-activated protein kinase (AMPK)-signaling pathway activity was assessed by ELISA and Western blotting, respectively. AMPK agonist (AICAR) was used to analyze the role of this kinase. The expression of K_ATP subunit (Kir6.1, Kir6.2, SUR1, SUR2A and SUR2B) was examined using quantitative real-time PCR (RT-PCR).

Results: We found that HG was significant decreases in the expression of Kir6.1, SUB2A and SUB2B, three subunits of K_ATP, TIMP-2 production, K_ATP channel activity and AMPK activity, while it promoted the cell proliferation and Collagen IV production in rat mesangial cells. Pretreatment with K_ATP selective opener (diazoxide, DZX) significantly inhibited HG-induced mesangial cell proliferation, Collagen IV production and decrease in K_ATP channel activity in rat mesangial cells, which were reversed by pretreatment of 5-hydroxydecanoate, a selective inhibitor of K_ATP. Moreover, AICAR pretreatment inhibited HG-induced decrease in K_ATP channel activity.

Conclusions: Taken together, activating AMPK-K_ATP signaling may protect against HG-induced mesangial cell proliferation and Collagen IV production, and, thereby, provides new insights into the molecular mechanisms underlying early diabetic nephropathy (DN).

Keywords: AMP-activated protein kinase; Cell proliferation; Collagen IV; High glucose; KATP channel.