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15-Deoxy-Delta-Prostaglandin J(2) Upregulates the Expression of LPS-Induced IL-8/CXCL8 mRNA in Vascular Smooth Muscle Cells from Spontaneously Hypertensive Rats.

Kim JH, Kim HS - Immune Netw (2009)

Bottom Line: As a result of these properties, we examined the effect of 15d-PGJ(2) on the LPS-induced IL-8/CXCL8 mRNA expression in VSMCs from SHR.However, inhibition of the p38 signaling pathway augmented the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 mRNA.Our results indicate that the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 expression in SHR VSMCs is mediated through the PPARgamma and ERK pathway, and may be related to NAD(P)H oxidase activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, College of Medicine, and Aging-associated Vascular Disease Research Center, Yeungnam University, Daegu, Korea.

ABSTRACT

Background: 15d-PGJ(2) has been known to act as an anti-inflammatory agent and has anti-hypertensive effects. As a result of these properties, we examined the effect of 15d-PGJ(2) on the LPS-induced IL-8/CXCL8 mRNA expression in VSMCs from SHR.

Methods: Effect and action mechanism of 15d-PGJ(2) on the expression of LPS-induced IL-8/CXCL8 mRNA in VSMCs from SHR and WKY were examined by using real-time polymerase chain reaction, electrophoretic mobility shift assay for NF-kappaB avtivity, Western blotting analysis for ERK and p38 phosphorylation and flow cytometry for NAD(P)H oxidase activity.

Results: 15d-PGJ(2) decreased the expression of LPS-induced IL-8/CXCL8 mRNA in WKY VSMCs, but increased the expression of LPS-induced IL-8/CXCL8 mRNA in SHR VSMCs. The upregulatory effect of 15d-PGJ(2) in SHR VSMCs was mediated through PPARgamma, and dependent on NF-kappaB activation and ERK phosphorylation. However, inhibition of the p38 signaling pathway augmented the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 mRNA. A NAD(P)H oxidase inhibitor inhibited the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs, and an increase in NAD(P)H oxidase activity was detected in SHR VSMCs treated with 15d-PGJ(2)/LPS.

Conclusion: Our results indicate that the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 expression in SHR VSMCs is mediated through the PPARgamma and ERK pathway, and may be related to NAD(P)H oxidase activity. However, p38 inactivation may also play an important role in 15d-PGJ(2)/LPS-induced IL-8/CXCL8 expression in SHR VSMCs.

No MeSH data available.


Related in: MedlinePlus

Activity of NAD(P)H oxidase mediates the upregulatory effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in SHR VSMCs. (A) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) in the absence or presence of DPI (10 µM) for 4 h. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with 15d-PGJ2/LPS. (B) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) for 4 h, stained with DCF-DA (50 µM) for ROS detection, and subjected to flow cytometry. Bars represent means±SD from four independent experiments. *p<0.05 vs. VSMCs treated with LPS alone.
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Figure 6: Activity of NAD(P)H oxidase mediates the upregulatory effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in SHR VSMCs. (A) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) in the absence or presence of DPI (10 µM) for 4 h. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with 15d-PGJ2/LPS. (B) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) for 4 h, stained with DCF-DA (50 µM) for ROS detection, and subjected to flow cytometry. Bars represent means±SD from four independent experiments. *p<0.05 vs. VSMCs treated with LPS alone.

Mentions: Real-time PCR was performed on VSMCs SHR after they were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) in the absence or presence of flavin containing oxidase inhibitor, DPI (10 µM) for 4 h. DPI remarkably decreased the expression of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA; thus, blocking the upregulatory effects of 15d-PGJ2 on LPS-induced IL-8/CXCL8 expression (Fig. 6A). To support these results, the ability of 15d-PGJ2/LPS to induce NAD(P)H oxidase activity was examined in SHR VSMCs. ROS generation in SHR VSMCs was measured by flow cytometric analysis of DCF-DA-stained VSMCs. DCF-DA fluorescence can be also used as a measurement of NAD(P)H oxidase activity. From this analysis, SHR VSMCs treated with 15d-PGJ2/LPS was shown to increase DCF-DA fluorescence slightly compared to those treated with LPS alone (Fig. 6B).


15-Deoxy-Delta-Prostaglandin J(2) Upregulates the Expression of LPS-Induced IL-8/CXCL8 mRNA in Vascular Smooth Muscle Cells from Spontaneously Hypertensive Rats.

Kim JH, Kim HS - Immune Netw (2009)

Activity of NAD(P)H oxidase mediates the upregulatory effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in SHR VSMCs. (A) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) in the absence or presence of DPI (10 µM) for 4 h. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with 15d-PGJ2/LPS. (B) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) for 4 h, stained with DCF-DA (50 µM) for ROS detection, and subjected to flow cytometry. Bars represent means±SD from four independent experiments. *p<0.05 vs. VSMCs treated with LPS alone.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Activity of NAD(P)H oxidase mediates the upregulatory effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in SHR VSMCs. (A) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) in the absence or presence of DPI (10 µM) for 4 h. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with 15d-PGJ2/LPS. (B) VSMCs were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) for 4 h, stained with DCF-DA (50 µM) for ROS detection, and subjected to flow cytometry. Bars represent means±SD from four independent experiments. *p<0.05 vs. VSMCs treated with LPS alone.
Mentions: Real-time PCR was performed on VSMCs SHR after they were untreated or treated with LPS (1 µg/ml) and/or 15d-PGJ2 (10 µM) in the absence or presence of flavin containing oxidase inhibitor, DPI (10 µM) for 4 h. DPI remarkably decreased the expression of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA; thus, blocking the upregulatory effects of 15d-PGJ2 on LPS-induced IL-8/CXCL8 expression (Fig. 6A). To support these results, the ability of 15d-PGJ2/LPS to induce NAD(P)H oxidase activity was examined in SHR VSMCs. ROS generation in SHR VSMCs was measured by flow cytometric analysis of DCF-DA-stained VSMCs. DCF-DA fluorescence can be also used as a measurement of NAD(P)H oxidase activity. From this analysis, SHR VSMCs treated with 15d-PGJ2/LPS was shown to increase DCF-DA fluorescence slightly compared to those treated with LPS alone (Fig. 6B).

Bottom Line: As a result of these properties, we examined the effect of 15d-PGJ(2) on the LPS-induced IL-8/CXCL8 mRNA expression in VSMCs from SHR.However, inhibition of the p38 signaling pathway augmented the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 mRNA.Our results indicate that the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 expression in SHR VSMCs is mediated through the PPARgamma and ERK pathway, and may be related to NAD(P)H oxidase activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, College of Medicine, and Aging-associated Vascular Disease Research Center, Yeungnam University, Daegu, Korea.

ABSTRACT

Background: 15d-PGJ(2) has been known to act as an anti-inflammatory agent and has anti-hypertensive effects. As a result of these properties, we examined the effect of 15d-PGJ(2) on the LPS-induced IL-8/CXCL8 mRNA expression in VSMCs from SHR.

Methods: Effect and action mechanism of 15d-PGJ(2) on the expression of LPS-induced IL-8/CXCL8 mRNA in VSMCs from SHR and WKY were examined by using real-time polymerase chain reaction, electrophoretic mobility shift assay for NF-kappaB avtivity, Western blotting analysis for ERK and p38 phosphorylation and flow cytometry for NAD(P)H oxidase activity.

Results: 15d-PGJ(2) decreased the expression of LPS-induced IL-8/CXCL8 mRNA in WKY VSMCs, but increased the expression of LPS-induced IL-8/CXCL8 mRNA in SHR VSMCs. The upregulatory effect of 15d-PGJ(2) in SHR VSMCs was mediated through PPARgamma, and dependent on NF-kappaB activation and ERK phosphorylation. However, inhibition of the p38 signaling pathway augmented the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 mRNA. A NAD(P)H oxidase inhibitor inhibited the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs, and an increase in NAD(P)H oxidase activity was detected in SHR VSMCs treated with 15d-PGJ(2)/LPS.

Conclusion: Our results indicate that the upregulatory effect of 15d-PGJ(2) on LPS-induced IL-8/CXCL8 expression in SHR VSMCs is mediated through the PPARgamma and ERK pathway, and may be related to NAD(P)H oxidase activity. However, p38 inactivation may also play an important role in 15d-PGJ(2)/LPS-induced IL-8/CXCL8 expression in SHR VSMCs.

No MeSH data available.


Related in: MedlinePlus