Effect of rosiglitazone on the secretion of chemokines in renal tubular epithelial cells stimulated by lipopolysaccharide and the possible mechanism
10.3760/cma.j.issn.1001-7097.2010.12.007
- VernacularTitle:罗格列酮对脂多糖诱导肾小管上皮细胞趋化因子分泌的影响及可能机制
- Author:
Ying LU
;
Qiao ZHOU
;
Fang ZHONG
;
Xu HAO
;
Cong LI
;
Weiming WANG
;
Nan CHEN
- Publication Type:Journal Article
- Keywords:
Lipopolysaccharides;
Interleukin-8;
Chemokine CCL2;
PPAR gamma;
Rosiglitazone;
Renal tubular epithelial cells;
NF-kappa B
- From:
Chinese Journal of Nephrology
2010;26(12):909-914
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effect and mechanism of rosiglitazone on chemokines secretion in renal tubular epithelial cells (HK-2) stimulated by lipopolysaccharide (LPS). Methods Cells were divided into four groups: control (CON), LPS (1 mg/L),rosiglitazone (10 μmol/L), rosiglitazone (10 μmol/L) +LPS (1 mg/L). MCP-1 and IL-8 expression was measured using real time PCR and ELISA. PPARγ was knockdown by RNAi to investigate whether the inhibitory effect of rosiglitazone was PPARγ-dependent or -independent. The NF-κB in nucleus was detected by Western blotting. The DNA binding activity of NF-κB was determined by electrophoretic mobility shift assay. Results Compared with CON group, the expressions of IL-8 and MCP-1 were increased by (4.30±0.45) and (4.80±1.29) times in mRNA level, (1.39±0.18)and (2.11 ±0.47) times in protein level, respectively, in LPS-stimulated HK-2 cells (P＜0.05).Application of rosiglitazone followed by LPS significantly reduced IL-8 and MCP-1 secretion compared with LPS group (decreasing by 66.37% and 71.88% in mRNA levels, while 41.68% and 47.87% in protein levels) (P＜0.05). In pcDNATM 6.2-GW/EmGFP-miPPARγ transfected cells, IL-8and MCP-1 only were decreased by 18.16% and 16.83% in mRNA level, while 11.39% and 11.86%% in protein level in rosiglitazone pretreated group, showing no significant difference compared with LPS group. Rosiglitazone did not block NF-κB nuclear translocation while significantly inhibiting the DNA binding activity of NF-κB. Conclusions Rosiglitazone inhibits the expressions of MCP-1 and IL-8 via a PPARγ-dependent mechanism in HK-2 cells, resulting from inhibition the DNA binding activity of NF-κB.
