title: Anti-proliferative actions of T-type calcium channel inhibition in Thy1 nephritis.
abstract: Aberrant proliferation of mesangial cells (MCs) is a key finding in progressive glomerular disease. TH1177 is a small molecule that has been shown to inhibit low-voltage activated T-type Ca(2+) channels (TCCs). The current study investigates the effect of TH1177 on MC proliferation in vitro and in vivo. The effect of Ca(2+) channel inhibition on primary rat MC proliferation in vitro was studied using the microculture tetrazolium assay and by measuring bromodeoxyuridine incorporation. In vivo, rats with Thy1 nephritis were treated with TH1177 or vehicle. Glomerular injury and average glomerular cell number were determined in a blinded fashion. Immunostaining for Ki-67 and phosphorylated ERK were also performed. The expression of TCC isoforms in healthy and diseased tissue was investigated using quantitative real-time PCR. TCC blockade caused a significant reduction in rat MC proliferation in vitro, whereas L-type inhibition had no effect. Treatment of Thy1 nephritis with TH1177 significantly reduced glomerular injury (P < 0.005) and caused a 49% reduction in glomerular cell number (P < 0.005) compared to the placebo. TH1177 also reduced Ki-67-positive and pERK-positive cells per glomerulus by 52% (P < 0.01 and P < 0.005, respectively). These results demonstrate that TH1177 inhibits MC proliferation in vitro and in vivo, supporting the hypothesis that TCC inhibition may be a useful strategy for studying and modifying MC proliferative responses to injury.
authors.affiliation: Department of Renal Medicine, King's College London, London, United Kingdom. andrea.cove-smith@kcl.ac.uk
authors.forename: Andrea
authors.initials: A
authors.lastname: Cove-Smith
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Mulgrew
authors.forename: Olena
authors.initials: O
authors.lastname: Rudyk
authors.forename: Neelanjana
authors.initials: N
authors.lastname: Dutt
authors.forename: Linda M
authors.initials: LM
authors.lastname: McLatchie
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Shattock
authors.forename: Bruce M
authors.initials: BM
authors.lastname: Hendry
mesh_terms: D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D020747:Calcium Channels, T-Type / Q000187:drug effects*; D003404:Creatinine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005921:Glomerulonephritis / Q000188:drug therapy* / Q000473:pathology; D007672:Kidney Cortex / Q000378:metabolism; D007678:Kidney Glomerulus / Q000187:drug effects / Q000473:pathology; D008297:Male; D050527:Mesangial Cells / Q000473:pathology*; D009532:Nickel / Q000494:pharmacology; D011507:Proteinuria / Q000139:chemically induced; D011759:Pyrrolidines / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D018800:Thy-1 Antigens / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology
chemical_list: D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; D011759:Pyrrolidines; C405269:TH 1177; D018800:Thy-1 Antigens; C022838:nickel chloride; D009532:Nickel; D003404:Creatinine; D014700:Verapamil