Limits...
L-cystathionine inhibits oxidized low density lipoprotein-induced THP-1-derived macrophage inflammatory cytokine monocyte chemoattractant protein-1 generation via the NF-κB pathway.

Zhu M, Du J, Liu AD, Holmberg L, Chen SY, Bu D, Tang C, Jin H - Sci Rep (2015)

Bottom Line: Compared with the ox-LDL group, 0.3 mmol/L and 1.0 mmol/L L-cystathionine significantly inhibited the expression of THP-1-derived macrophage MCP-1.Mechanistically, 0.3 mmol/L and 1.0 mmol/L L-cystathionine suppressed phosphorylation and nuclear translocation of the NF-κB p65 protein, as well as the DNA binding activity and DNA binding level of NF-κB with the MCP-1 promoter, which resulted in a reduced THP-1-derived macrophage MCP-1 generation.This study suggests that L-cystathionine could inhibit the expression of MCP-1 in THP-1-derived macrophages induced by ox-LDL via inhibition of NF-κB p65 phosphorylation, nuclear translocation, and binding of the MCP-1 promoter sequence after entry into the nucleus.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, P. R. China.

ABSTRACT
This study aimed to explore whether and how L-cystathionine had any regulatory effect on the inflammatory response in THP-1-derived macrophages cultured in vitro under oxidized low-density lipoprotein (ox-LDL) stimulation. The human monocyte line THP-1 cell was cultured in vitro and differentiated into macrophages after 24 hours of PMA induction. Macrophages were pretreated with L-cystathionine and then treated with ox-LDL. The results showed that compared with the controls, ox-LDL stimulation significantly upregulated the expression of THP-1-derived macrophage MCP-1 by enhancing NF-κB p65 phosphorylation, nuclear translocation and DNA binding with the MCP-1 promoter. Compared with the ox-LDL group, 0.3 mmol/L and 1.0 mmol/L L-cystathionine significantly inhibited the expression of THP-1-derived macrophage MCP-1. Mechanistically, 0.3 mmol/L and 1.0 mmol/L L-cystathionine suppressed phosphorylation and nuclear translocation of the NF-κB p65 protein, as well as the DNA binding activity and DNA binding level of NF-κB with the MCP-1 promoter, which resulted in a reduced THP-1-derived macrophage MCP-1 generation. This study suggests that L-cystathionine could inhibit the expression of MCP-1 in THP-1-derived macrophages induced by ox-LDL via inhibition of NF-κB p65 phosphorylation, nuclear translocation, and binding of the MCP-1 promoter sequence after entry into the nucleus.

No MeSH data available.


Related in: MedlinePlus

L-cystathionine inhibited the phosphorylation of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage. The cells were pretreated with L-cystathionine (0.1 mmol/L, 0.3 mmol/L or 1.0 mmol/L, respectively) for 30 min, and were then stimulated with 50 mg/L ox-LDL for 30 min. (A) The effect of L-cystathionine on phosphorylation level of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage detected by western blot. (B) The basal phosphorylation level of NF-κB p65 protein in the THP-1-derived macrophage without ox-LDL treatment detected by western blot. L-Cth: L-cystathionine. **P < 0.01 compared with control group, ##p < 0.01 compared with ox-LDL group. Data are presented as mean ± SD (n = 3) of three independent experiments performed in triplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4447071&req=5

f2: L-cystathionine inhibited the phosphorylation of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage. The cells were pretreated with L-cystathionine (0.1 mmol/L, 0.3 mmol/L or 1.0 mmol/L, respectively) for 30 min, and were then stimulated with 50 mg/L ox-LDL for 30 min. (A) The effect of L-cystathionine on phosphorylation level of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage detected by western blot. (B) The basal phosphorylation level of NF-κB p65 protein in the THP-1-derived macrophage without ox-LDL treatment detected by western blot. L-Cth: L-cystathionine. **P < 0.01 compared with control group, ##p < 0.01 compared with ox-LDL group. Data are presented as mean ± SD (n = 3) of three independent experiments performed in triplicate.

Mentions: To begin, we studied whether phosphorylation levels of NF-κB p65 was involved in MCP-1 expression by L-cystathionine. A western blot test was used to detect the phosphorylation levels of NF-κB p65. THP-1-derived macrophages were pretreated with L-cystathionine for 30 min, and then were stimulated with 50 mg/L ox-LDL for 30 min. The results show that ox-LDL significantly enhanced phosphorylation levels of the THP-1-derived macrophage NF-κB p65 protein. Meanwhile, L-cystathionine dose-dependently declined phosphorylation levels of NF-κB p65 protein (Fig. 2A). However, L-cystathionine had no effect on the phosphorylation levels of the NF-κB p65 protein without ox-LDL stimulation (Fig. 2B).


L-cystathionine inhibits oxidized low density lipoprotein-induced THP-1-derived macrophage inflammatory cytokine monocyte chemoattractant protein-1 generation via the NF-κB pathway.

Zhu M, Du J, Liu AD, Holmberg L, Chen SY, Bu D, Tang C, Jin H - Sci Rep (2015)

L-cystathionine inhibited the phosphorylation of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage. The cells were pretreated with L-cystathionine (0.1 mmol/L, 0.3 mmol/L or 1.0 mmol/L, respectively) for 30 min, and were then stimulated with 50 mg/L ox-LDL for 30 min. (A) The effect of L-cystathionine on phosphorylation level of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage detected by western blot. (B) The basal phosphorylation level of NF-κB p65 protein in the THP-1-derived macrophage without ox-LDL treatment detected by western blot. L-Cth: L-cystathionine. **P < 0.01 compared with control group, ##p < 0.01 compared with ox-LDL group. Data are presented as mean ± SD (n = 3) of three independent experiments performed in triplicate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4447071&req=5

f2: L-cystathionine inhibited the phosphorylation of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage. The cells were pretreated with L-cystathionine (0.1 mmol/L, 0.3 mmol/L or 1.0 mmol/L, respectively) for 30 min, and were then stimulated with 50 mg/L ox-LDL for 30 min. (A) The effect of L-cystathionine on phosphorylation level of NF-κB p65 protein induced by ox-LDL in the THP-1-derived macrophage detected by western blot. (B) The basal phosphorylation level of NF-κB p65 protein in the THP-1-derived macrophage without ox-LDL treatment detected by western blot. L-Cth: L-cystathionine. **P < 0.01 compared with control group, ##p < 0.01 compared with ox-LDL group. Data are presented as mean ± SD (n = 3) of three independent experiments performed in triplicate.
Mentions: To begin, we studied whether phosphorylation levels of NF-κB p65 was involved in MCP-1 expression by L-cystathionine. A western blot test was used to detect the phosphorylation levels of NF-κB p65. THP-1-derived macrophages were pretreated with L-cystathionine for 30 min, and then were stimulated with 50 mg/L ox-LDL for 30 min. The results show that ox-LDL significantly enhanced phosphorylation levels of the THP-1-derived macrophage NF-κB p65 protein. Meanwhile, L-cystathionine dose-dependently declined phosphorylation levels of NF-κB p65 protein (Fig. 2A). However, L-cystathionine had no effect on the phosphorylation levels of the NF-κB p65 protein without ox-LDL stimulation (Fig. 2B).

Bottom Line: Compared with the ox-LDL group, 0.3 mmol/L and 1.0 mmol/L L-cystathionine significantly inhibited the expression of THP-1-derived macrophage MCP-1.Mechanistically, 0.3 mmol/L and 1.0 mmol/L L-cystathionine suppressed phosphorylation and nuclear translocation of the NF-κB p65 protein, as well as the DNA binding activity and DNA binding level of NF-κB with the MCP-1 promoter, which resulted in a reduced THP-1-derived macrophage MCP-1 generation.This study suggests that L-cystathionine could inhibit the expression of MCP-1 in THP-1-derived macrophages induced by ox-LDL via inhibition of NF-κB p65 phosphorylation, nuclear translocation, and binding of the MCP-1 promoter sequence after entry into the nucleus.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, P. R. China.

ABSTRACT
This study aimed to explore whether and how L-cystathionine had any regulatory effect on the inflammatory response in THP-1-derived macrophages cultured in vitro under oxidized low-density lipoprotein (ox-LDL) stimulation. The human monocyte line THP-1 cell was cultured in vitro and differentiated into macrophages after 24 hours of PMA induction. Macrophages were pretreated with L-cystathionine and then treated with ox-LDL. The results showed that compared with the controls, ox-LDL stimulation significantly upregulated the expression of THP-1-derived macrophage MCP-1 by enhancing NF-κB p65 phosphorylation, nuclear translocation and DNA binding with the MCP-1 promoter. Compared with the ox-LDL group, 0.3 mmol/L and 1.0 mmol/L L-cystathionine significantly inhibited the expression of THP-1-derived macrophage MCP-1. Mechanistically, 0.3 mmol/L and 1.0 mmol/L L-cystathionine suppressed phosphorylation and nuclear translocation of the NF-κB p65 protein, as well as the DNA binding activity and DNA binding level of NF-κB with the MCP-1 promoter, which resulted in a reduced THP-1-derived macrophage MCP-1 generation. This study suggests that L-cystathionine could inhibit the expression of MCP-1 in THP-1-derived macrophages induced by ox-LDL via inhibition of NF-κB p65 phosphorylation, nuclear translocation, and binding of the MCP-1 promoter sequence after entry into the nucleus.

No MeSH data available.


Related in: MedlinePlus