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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.


Effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in VSMCs from SHR and WKY, and the time course of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs. (A) VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or/and 15d-PGJ2 (10 µM) for 4 h, and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with LPS alone. (B) SHR VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or LPS plus 15d-PGJ2 (10 µM) simultaneously (15d-PGJ2/LPS) for the indicated times and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments.
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Figure 1: Effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in VSMCs from SHR and WKY, and the time course of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs. (A) VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or/and 15d-PGJ2 (10 µM) for 4 h, and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with LPS alone. (B) SHR VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or LPS plus 15d-PGJ2 (10 µM) simultaneously (15d-PGJ2/LPS) for the indicated times and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments.

Mentions: We examined the differential effect of LPS on IL-8/CXCL8 mRNA expression in SHR VSMCs in comparison to WKY VSMCs. From this experiment, we found that the expression of LPS-induced IL-8/CXCL8 mRNA was greater in SHR VSMCs than WKY. Real-time PCR was performed on VSMCs after they were untreated (NT) or treated with LPS (1 µg/ml), 15d-PGJ2 (10 µM) or LPS plus 15d-PGJ2 simultaneously (15d-PGJ2/LPS) for 4 h. 15d-PGJ2 treatment had different effects on SHR VSMCs relative to WKY VSMCs, where 15d-PGJ2 had upregulatory effect on LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs and suppressive effect on LPS-induced IL-8/CXCL8 mRNA expression in WKY VSMCs. 15d-PGJ2 alone did not induce IL-8/CXCL8 mRNA expression in SHR VSMCs significantly (Fig. 1A). The time course of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA expression was determined in SHR VSMCs over a 0 to 8 h time period. In this experiment, we found that the expression of IL-8/CXCL8 mRNA induced by 15d-PGJ2/LPS was almost the same as that for cells treated with LPS alone until 2 h after treatment. However, the expression levels of IL-8/CXCL8 mRNA induced by 15d-PGJ2/LPS were significantly greater than those in the cells treated with LPS alone from the 4 h period (Fig. 1B).


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)

Effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in VSMCs from SHR and WKY, and the time course of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs. (A) VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or/and 15d-PGJ2 (10 µM) for 4 h, and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with LPS alone. (B) SHR VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or LPS plus 15d-PGJ2 (10 µM) simultaneously (15d-PGJ2/LPS) for the indicated times and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments.
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Figure 1: Effect of 15d-PGJ2 on the expression of LPS-induced IL-8/CXCL8 mRNA in VSMCs from SHR and WKY, and the time course of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs. (A) VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or/and 15d-PGJ2 (10 µM) for 4 h, and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments. *p<0.05 vs. VSMCs treated with LPS alone. (B) SHR VSMCs were untreated (NT) or treated with LPS (1 µg/ml) or LPS plus 15d-PGJ2 (10 µM) simultaneously (15d-PGJ2/LPS) for the indicated times and the total RNA was analyzed by real-time PCR. Bars represent means±SD from three independent experiments.
Mentions: We examined the differential effect of LPS on IL-8/CXCL8 mRNA expression in SHR VSMCs in comparison to WKY VSMCs. From this experiment, we found that the expression of LPS-induced IL-8/CXCL8 mRNA was greater in SHR VSMCs than WKY. Real-time PCR was performed on VSMCs after they were untreated (NT) or treated with LPS (1 µg/ml), 15d-PGJ2 (10 µM) or LPS plus 15d-PGJ2 simultaneously (15d-PGJ2/LPS) for 4 h. 15d-PGJ2 treatment had different effects on SHR VSMCs relative to WKY VSMCs, where 15d-PGJ2 had upregulatory effect on LPS-induced IL-8/CXCL8 mRNA expression in SHR VSMCs and suppressive effect on LPS-induced IL-8/CXCL8 mRNA expression in WKY VSMCs. 15d-PGJ2 alone did not induce IL-8/CXCL8 mRNA expression in SHR VSMCs significantly (Fig. 1A). The time course of 15d-PGJ2/LPS-induced IL-8/CXCL8 mRNA expression was determined in SHR VSMCs over a 0 to 8 h time period. In this experiment, we found that the expression of IL-8/CXCL8 mRNA induced by 15d-PGJ2/LPS was almost the same as that for cells treated with LPS alone until 2 h after treatment. However, the expression levels of IL-8/CXCL8 mRNA induced by 15d-PGJ2/LPS were significantly greater than those in the cells treated with LPS alone from the 4 h period (Fig. 1B).

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.