Regulation of inflammatory response in human chondrocytes by lentiviral mediated RNA interference against S100A10.
OBJECTIVE
The aim of the present study was to evaluate the effects of S100A10 silencing on the inflammatory response in human chondrocytes (HCs).The inflammation induced by lipopolysaccharide (LPS) was investigated in HCs in which the S100A10 was blocked with a lentiviral shRNA vector.


METHODS
A lentiviral shRNA vector targeting S100A10 was constructed and packaged to effectively block S100A10 expression in HCs. HCs were infected with the lentivirus. S100A10 expression levels in HCs were detected by western blot analysis. Enzyme-linked immunosorbent assay (ELISA) was employed to evaluate the change of cytokine secretion levels. The effects of S100A10 silencing on the activation of mitogen-activated protein kinases (MAPKs) and NF-κB signaling pathway were also determined by western blot analysis. In addition, fluo-3-AM was used to demonstrate the change in calcium mobilization.


RESULTS
Lentivirus effectively infected the HCs and inhibited the expression of S100A10. HCs with downregulated S100A10 showed significantly decreased production of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-10. S100A10 silencing markedly suppressed the activation of MAPKs induced by LPS. Furthermore, the calcium concentration increase in HCs stimulated by LPS was also inhibited by S100A10 knockdown.


CONCLUSIONS
Our investigation demonstrated that S100A10 might be considered as a potential target for anti-inflammatory treatment.
Department of Orthopaedics, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Changzhi
C
Song
Xiaoye
X
Zhou
Qirong
Q
Dong
Rengen
R
Fan
Guangzhou
G
Wu
Biao
B
Ji
Qingbing
Q
Meng
Minqian
M
Zheng
D000328:Adult; D017306:Annexin A2 / Q000235:genetics*; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics; D008070:Lipopolysaccharides; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D034622:RNA Interference; D009418:S100 Proteins / Q000235:genetics*; D055815:Young Adult; D000099248:S100 Calcium Binding Protein A10
D017306:Annexin A2; D002118:Calcium; D016207:Cytokines; D008070:Lipopolysaccharides; D020928:Mitogen-Activated Protein Kinases; D009418:S100 Proteins; D000099248:S100 Calcium Binding Protein A10