title: Regulation of TRPC6 Channels by Non-Steroidal Anti-Inflammatory Drugs.
abstract: Family focal segmental glomerulosclerosis (FSGS) is characterized by sclerosis and hyalinosis of particular loops of glomeruli and is one of the causes of the nephrotic syndrome. Certain mutations in the structure of TRPC6 channels are the genetic impetus for FSGS development resulting in podocytes functional abnormalities and various nephropathies. We have recently demonstrated that non-steroid anti-inflammatory drugs (NSAID) ibuprofen and diclofenac decrease the activity of endogenous TRPC like cal cium channels in the podocytes of the freshly isolated rat glomeruli. It has also been shown that TRPC6 chan nels are expressed in the podocytes. In the current study we have functionally reconstituted TRPC6 channels in mammalian cells to investigate the effects of diclofenac on the activity of wild type TRPC6 channel and TRPC6P112Qchannel containing a mutation in the N-terminus that was described in FSGS patients. Intracellular calcium level measurements in transfected cells revealed a more intensive carbachol induced increase of calcium concentration in HEK 293 cells expressing TRPC6P112Qversus the cells expressing wild-type TRPC6. We also performed patch-clamp experiments to study TRPC6 channels reconstituted in Chinese hamster ovary (CHO) cell line and found that application of diclofenac (500 μM) acutely reduced single channel activity. Preincubation with diclofenac (100 μM) also decreased the whole cell current in CHO cells overexpressing TRPC6P112Q. Therefore, our previously published data on the effects of NSAID on TRPC-like channels in the isolated rat glomeruli, along with this current investigation on the cultured overexpressed mammalian cells, allows hypothesizing that TRPC6 channels may be a target for NSAID that can be impor tant in the treatment of FSGS.
authors.affiliation: Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI 53226, USA.|Institute of Cytology, Russian Academy of Sciences, Tikhoretskii pr. 4, St. Petersburg, 194064 Russia.
authors.forename: D V
authors.initials: DV
authors.lastname: Ilatovskaya
authors.affiliation: Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI 53226, USA.
authors.forename: T S
authors.initials: TS
authors.lastname: Pavlov
authors.affiliation: Institute of Cytology, Russian Academy of Sciences, Tikhoretskii pr. 4, St. Petersburg, 194064 Russia.
authors.forename: Y A
authors.initials: YA
authors.lastname: Negulyaev
authors.affiliation: Medical College of Wisconsin, 8701 Watertown Plank Rd., Milwaukee, WI 53226, USA.
authors.forename: A
authors.initials: A
authors.lastname: Staruschenko
keywords: TRPC6; calcium; focal segmental glomerulosclerosis; ion channels; non-steroid anti-inflammatory drugs