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Atorvastatin increases lipopolysaccharide-induced expression of tumour necrosis factor-α-induced protein 8-like 2 in RAW264.7 cells.

Liu MW, Su MX, Zhang W, Wang L, Qian CY - Exp Ther Med (2014)

Bottom Line: The cells were harvested, and the mRNA and protein expression levels of TIPE2, macrophage migration inhibitory factor (MIF), IκB and nuclear factor (NF-κB)-κB were analysed using quantitative polymerase chain reaction and western blotting analysis, respectively, the expression of NOS, COX-2 and HO-1 protein were essayed by western blotting analysis, NO and ROS activities were determined.Meanwhile, LPS enhanced the expression of NOS and COX-2 protein, blocked HO-1 protein expression, increased NO and PGE2 production and ROS activity in a dose dependent manner in RAW264.7 cells.Atorvastatin significantly increased LPS induced expression of TIPE2, downregulated the expression of NOS, COX-2, MIF and NF-κB and the production of PGE2, NO, IL-6 and TNF-α in a time and dose dependent manner, and increased HO-1 protein expression, reduced ROS production in a dose dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650032, P.R. China.

ABSTRACT
RAW264.7 cells are one of the major sources of productive inflammatory biomediators, including tumour necrosis factor-α (TNF-α) and interleukin (IL)-6. TNF-α-induced protein 8-like 2 (TIPE2) is an essential negative regulator of Toll-like and T-cell receptors, and the selective expression in the immune system prevents hyper-responsiveness and maintains immune homeostasis. The aim of the present study was to investigate whether atorvastatin upregulates the expression of TIPE2 and further regulates the inflammatory response and oxidation emergency response in RAW264.7 cells. RAW264.7 cells were incubated in Dulbecco's modified Eagle's medium containing lipopolysaccharide (LPS) in the presence or absence of atorvastatin. Following incubation, the medium was collected and the levels of TNF-α and IL-6 were measured using an enzyme-linked immunosorbent assay. The cells were harvested, and the mRNA and protein expression levels of TIPE2, macrophage migration inhibitory factor (MIF), IκB and nuclear factor (NF-κB)-κB were analysed using quantitative polymerase chain reaction and western blotting analysis, respectively, the expression of NOS, COX-2 and HO-1 protein were essayed by western blotting analysis, NO and ROS activities were determined. The results revealed that LPS increased the mRNA and protein expression levels of TIPE2, MIF and NF-κB, as well as the production of IL-6 and TNF-α, in a dose and time dependent manner in RAW264.7 cells. Meanwhile, LPS enhanced the expression of NOS and COX-2 protein, blocked HO-1 protein expression, increased NO and PGE2 production and ROS activity in a dose dependent manner in RAW264.7 cells. Atorvastatin significantly increased LPS induced expression of TIPE2, downregulated the expression of NOS, COX-2, MIF and NF-κB and the production of PGE2, NO, IL-6 and TNF-α in a time and dose dependent manner, and increased HO-1 protein expression, reduced ROS production in a dose dependent manner. The observations indicated that atorvastatin upregulated LPS induced expression of TIPE2 and consequently inhibited MIF, NF-κB, NOS and COX-2 expression and the production of NO, PGE2, TNF-α and IL-6, increased HO-1 expression, and inhibited ROS activity in cultured RAW264.7 cells.

No MeSH data available.


Related in: MedlinePlus

RAW264.7 cells were incubated with various concentrations of atorvastatin for 9 h in the presence of 10 ng/ml LPS. HO-1 and ROS expression levels were measured. (A) Representative western blot shows the protein expression levels of HO-1 in RAW264.7 cells. Statistical summary of the densitometric analysis of (B) HO-1 protein expression and (C) ROS production in RAW264.7 cells. Data are expressed as the mean ± standard error of the mean of three independent experiments.*P<0.05 and **P<0.01, vs. atorvastatin (10, 15 and 20 μM). LPS, lipopolysaccharide; ROS, reactive oxygen species; HO-1, haem oxygenase; a, LPS (10 ng/ml); b, LPS (10 ng/ml) + atorvastatin (10 μM); c, LPS (10 ng/ml) + atorvastatin (15 μM); d, LPS (10 ng/ml) + atorvastatin (20 μM).
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f9-etm-08-01-0219: RAW264.7 cells were incubated with various concentrations of atorvastatin for 9 h in the presence of 10 ng/ml LPS. HO-1 and ROS expression levels were measured. (A) Representative western blot shows the protein expression levels of HO-1 in RAW264.7 cells. Statistical summary of the densitometric analysis of (B) HO-1 protein expression and (C) ROS production in RAW264.7 cells. Data are expressed as the mean ± standard error of the mean of three independent experiments.*P<0.05 and **P<0.01, vs. atorvastatin (10, 15 and 20 μM). LPS, lipopolysaccharide; ROS, reactive oxygen species; HO-1, haem oxygenase; a, LPS (10 ng/ml); b, LPS (10 ng/ml) + atorvastatin (10 μM); c, LPS (10 ng/ml) + atorvastatin (15 μM); d, LPS (10 ng/ml) + atorvastatin (20 μM).

Mentions: To investigate whether atorvastatin affected LPS-induced HO-1 and ROS expression, RAW264.7 cells were incubated with various concentrations of atorvastatin for 9 h in the presence of 10 ng/ml LPS. As shown in Fig. 9, atorvastatin upregulated HO-1 expression, but reduced the production of ROS in a dose-dependent manner.


Atorvastatin increases lipopolysaccharide-induced expression of tumour necrosis factor-α-induced protein 8-like 2 in RAW264.7 cells.

Liu MW, Su MX, Zhang W, Wang L, Qian CY - Exp Ther Med (2014)

RAW264.7 cells were incubated with various concentrations of atorvastatin for 9 h in the presence of 10 ng/ml LPS. HO-1 and ROS expression levels were measured. (A) Representative western blot shows the protein expression levels of HO-1 in RAW264.7 cells. Statistical summary of the densitometric analysis of (B) HO-1 protein expression and (C) ROS production in RAW264.7 cells. Data are expressed as the mean ± standard error of the mean of three independent experiments.*P<0.05 and **P<0.01, vs. atorvastatin (10, 15 and 20 μM). LPS, lipopolysaccharide; ROS, reactive oxygen species; HO-1, haem oxygenase; a, LPS (10 ng/ml); b, LPS (10 ng/ml) + atorvastatin (10 μM); c, LPS (10 ng/ml) + atorvastatin (15 μM); d, LPS (10 ng/ml) + atorvastatin (20 μM).
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f9-etm-08-01-0219: RAW264.7 cells were incubated with various concentrations of atorvastatin for 9 h in the presence of 10 ng/ml LPS. HO-1 and ROS expression levels were measured. (A) Representative western blot shows the protein expression levels of HO-1 in RAW264.7 cells. Statistical summary of the densitometric analysis of (B) HO-1 protein expression and (C) ROS production in RAW264.7 cells. Data are expressed as the mean ± standard error of the mean of three independent experiments.*P<0.05 and **P<0.01, vs. atorvastatin (10, 15 and 20 μM). LPS, lipopolysaccharide; ROS, reactive oxygen species; HO-1, haem oxygenase; a, LPS (10 ng/ml); b, LPS (10 ng/ml) + atorvastatin (10 μM); c, LPS (10 ng/ml) + atorvastatin (15 μM); d, LPS (10 ng/ml) + atorvastatin (20 μM).
Mentions: To investigate whether atorvastatin affected LPS-induced HO-1 and ROS expression, RAW264.7 cells were incubated with various concentrations of atorvastatin for 9 h in the presence of 10 ng/ml LPS. As shown in Fig. 9, atorvastatin upregulated HO-1 expression, but reduced the production of ROS in a dose-dependent manner.

Bottom Line: The cells were harvested, and the mRNA and protein expression levels of TIPE2, macrophage migration inhibitory factor (MIF), IκB and nuclear factor (NF-κB)-κB were analysed using quantitative polymerase chain reaction and western blotting analysis, respectively, the expression of NOS, COX-2 and HO-1 protein were essayed by western blotting analysis, NO and ROS activities were determined.Meanwhile, LPS enhanced the expression of NOS and COX-2 protein, blocked HO-1 protein expression, increased NO and PGE2 production and ROS activity in a dose dependent manner in RAW264.7 cells.Atorvastatin significantly increased LPS induced expression of TIPE2, downregulated the expression of NOS, COX-2, MIF and NF-κB and the production of PGE2, NO, IL-6 and TNF-α in a time and dose dependent manner, and increased HO-1 protein expression, reduced ROS production in a dose dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Department of Emergency, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650032, P.R. China.

ABSTRACT
RAW264.7 cells are one of the major sources of productive inflammatory biomediators, including tumour necrosis factor-α (TNF-α) and interleukin (IL)-6. TNF-α-induced protein 8-like 2 (TIPE2) is an essential negative regulator of Toll-like and T-cell receptors, and the selective expression in the immune system prevents hyper-responsiveness and maintains immune homeostasis. The aim of the present study was to investigate whether atorvastatin upregulates the expression of TIPE2 and further regulates the inflammatory response and oxidation emergency response in RAW264.7 cells. RAW264.7 cells were incubated in Dulbecco's modified Eagle's medium containing lipopolysaccharide (LPS) in the presence or absence of atorvastatin. Following incubation, the medium was collected and the levels of TNF-α and IL-6 were measured using an enzyme-linked immunosorbent assay. The cells were harvested, and the mRNA and protein expression levels of TIPE2, macrophage migration inhibitory factor (MIF), IκB and nuclear factor (NF-κB)-κB were analysed using quantitative polymerase chain reaction and western blotting analysis, respectively, the expression of NOS, COX-2 and HO-1 protein were essayed by western blotting analysis, NO and ROS activities were determined. The results revealed that LPS increased the mRNA and protein expression levels of TIPE2, MIF and NF-κB, as well as the production of IL-6 and TNF-α, in a dose and time dependent manner in RAW264.7 cells. Meanwhile, LPS enhanced the expression of NOS and COX-2 protein, blocked HO-1 protein expression, increased NO and PGE2 production and ROS activity in a dose dependent manner in RAW264.7 cells. Atorvastatin significantly increased LPS induced expression of TIPE2, downregulated the expression of NOS, COX-2, MIF and NF-κB and the production of PGE2, NO, IL-6 and TNF-α in a time and dose dependent manner, and increased HO-1 protein expression, reduced ROS production in a dose dependent manner. The observations indicated that atorvastatin upregulated LPS induced expression of TIPE2 and consequently inhibited MIF, NF-κB, NOS and COX-2 expression and the production of NO, PGE2, TNF-α and IL-6, increased HO-1 expression, and inhibited ROS activity in cultured RAW264.7 cells.

No MeSH data available.


Related in: MedlinePlus