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The PPARδ ligand GW501516 reduces growth but not apoptosis in mouse inner medullary collecting duct cells.

Clark J, Nasrallah R, Hébert RL - PPAR Res (2009)

Bottom Line: The collecting duct (CD) expresses considerable amounts of PPARδ.High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability.Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, Kidney Research Centre, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada K1H 8M5.

ABSTRACT
The collecting duct (CD) expresses considerable amounts of PPARδ. While its role is unknown in the CD, in other renal cells it has been shown to regulate both growth and apoptosis. We thus hypothesized that PPARδ reduces apoptotic responses and stimulates cell growth in the mouse CD, and examined the effect of GW501516, a synthetic PPARδ ligand, on these responses in mouse IMCD-K2 cells. High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability. Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels. These results show that a PPARδ ligand inhibits growth but does not affect anisomycin-apoptosis in a mouse IMCD cell line. This could have therapeutic implications for renal diseases associated with increased CD growth responses.

No MeSH data available.


Related in: MedlinePlus

Pretreatment with GW501516 doesnot significantly alter anisomycin-induced cleaved caspase-3 protein in IMCD-K2cells. Protein was isolated from IMCD-K2 cellsthat had been treated with control serum-free media, 10−6 or 10−8 M GW501516, 250 ng/mL anisomycin (A250 or A) for 24 hours, 10−6 MGW501516 for 3 hours pretreatment followed by 10−6 M GW501516 for 24hours with A250 (GW-6/A250), or 10−8 M GW501516 for 3 hourspretreatment followed by 10−8 M GW501516 for 24 hours with A250(GW-8/A250). (a) The lysates were run on an SDS-PAGE gel and incubated with ananticleaved caspase-3 antibody (1 : 1000). (b) The membranes were stripped and incubated with anti-β-actinantibody (1 : 10000) to normalize for densitometry. Expression is presented asfold A250. Values are mean  ±   S.E.M.;  n = 8.
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fig10: Pretreatment with GW501516 doesnot significantly alter anisomycin-induced cleaved caspase-3 protein in IMCD-K2cells. Protein was isolated from IMCD-K2 cellsthat had been treated with control serum-free media, 10−6 or 10−8 M GW501516, 250 ng/mL anisomycin (A250 or A) for 24 hours, 10−6 MGW501516 for 3 hours pretreatment followed by 10−6 M GW501516 for 24hours with A250 (GW-6/A250), or 10−8 M GW501516 for 3 hourspretreatment followed by 10−8 M GW501516 for 24 hours with A250(GW-8/A250). (a) The lysates were run on an SDS-PAGE gel and incubated with ananticleaved caspase-3 antibody (1 : 1000). (b) The membranes were stripped and incubated with anti-β-actinantibody (1 : 10000) to normalize for densitometry. Expression is presented asfold A250. Values are mean ± S.E.M.; n = 8.

Mentions: Activationof caspase was also measured by Western Blotting using a cleaved caspase-3antibody. As shown in Figure 10(a), a 17 kDa band corresponding to cleavedcaspase-3 was observed for the lysates that had been treated with anisomycin. The intensity of the bands in the lanes representing control and GW501516 alonewas too low to be analyzed in the majority of the experiments; therefore,densitometric analysis was not performed for those samples. The levels ofactive caspase-3 (see Figure 10(b)) increased to 1.48 and 1.18 of anisomycinalone with both cotreatments (10−6 and 10−8 M GW501516,resp.) but the difference was not significant.


The PPARδ ligand GW501516 reduces growth but not apoptosis in mouse inner medullary collecting duct cells.

Clark J, Nasrallah R, Hébert RL - PPAR Res (2009)

Pretreatment with GW501516 doesnot significantly alter anisomycin-induced cleaved caspase-3 protein in IMCD-K2cells. Protein was isolated from IMCD-K2 cellsthat had been treated with control serum-free media, 10−6 or 10−8 M GW501516, 250 ng/mL anisomycin (A250 or A) for 24 hours, 10−6 MGW501516 for 3 hours pretreatment followed by 10−6 M GW501516 for 24hours with A250 (GW-6/A250), or 10−8 M GW501516 for 3 hourspretreatment followed by 10−8 M GW501516 for 24 hours with A250(GW-8/A250). (a) The lysates were run on an SDS-PAGE gel and incubated with ananticleaved caspase-3 antibody (1 : 1000). (b) The membranes were stripped and incubated with anti-β-actinantibody (1 : 10000) to normalize for densitometry. Expression is presented asfold A250. Values are mean  ±   S.E.M.;  n = 8.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig10: Pretreatment with GW501516 doesnot significantly alter anisomycin-induced cleaved caspase-3 protein in IMCD-K2cells. Protein was isolated from IMCD-K2 cellsthat had been treated with control serum-free media, 10−6 or 10−8 M GW501516, 250 ng/mL anisomycin (A250 or A) for 24 hours, 10−6 MGW501516 for 3 hours pretreatment followed by 10−6 M GW501516 for 24hours with A250 (GW-6/A250), or 10−8 M GW501516 for 3 hourspretreatment followed by 10−8 M GW501516 for 24 hours with A250(GW-8/A250). (a) The lysates were run on an SDS-PAGE gel and incubated with ananticleaved caspase-3 antibody (1 : 1000). (b) The membranes were stripped and incubated with anti-β-actinantibody (1 : 10000) to normalize for densitometry. Expression is presented asfold A250. Values are mean ± S.E.M.; n = 8.
Mentions: Activationof caspase was also measured by Western Blotting using a cleaved caspase-3antibody. As shown in Figure 10(a), a 17 kDa band corresponding to cleavedcaspase-3 was observed for the lysates that had been treated with anisomycin. The intensity of the bands in the lanes representing control and GW501516 alonewas too low to be analyzed in the majority of the experiments; therefore,densitometric analysis was not performed for those samples. The levels ofactive caspase-3 (see Figure 10(b)) increased to 1.48 and 1.18 of anisomycinalone with both cotreatments (10−6 and 10−8 M GW501516,resp.) but the difference was not significant.

Bottom Line: The collecting duct (CD) expresses considerable amounts of PPARδ.High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability.Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, Kidney Research Centre, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada K1H 8M5.

ABSTRACT
The collecting duct (CD) expresses considerable amounts of PPARδ. While its role is unknown in the CD, in other renal cells it has been shown to regulate both growth and apoptosis. We thus hypothesized that PPARδ reduces apoptotic responses and stimulates cell growth in the mouse CD, and examined the effect of GW501516, a synthetic PPARδ ligand, on these responses in mouse IMCD-K2 cells. High doses of GW501516 decreased both DNA and protein synthesis in these cells by 80%, but had no overall effect on cell viability. Although anisomycin treatment resulted in an increase of caspase-3 levels of about 2.59-fold of control, GW501516 did not affect anisomycin-induced changes in active caspase-3 levels. These results show that a PPARδ ligand inhibits growth but does not affect anisomycin-apoptosis in a mouse IMCD cell line. This could have therapeutic implications for renal diseases associated with increased CD growth responses.

No MeSH data available.


Related in: MedlinePlus