Knockdown of low density lipoprotein receptor-related protein 1 in chondrocytes
10.3969/j.issn.2095-4344.2016.15.006
- VernacularTitle:软骨细胞膜蛋白低密度脂蛋白受体相关蛋白1低表达的意义
- Author:
Erping YANG
;
Fei PENG
;
Jie LIANG
;
Yuanli DU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2016;20(15):2171-2177
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Tumor necrosis factor α, as a pathogenic factor, induces the inflammatory reaction mainlyvia the activation of the nuclear factor kappa B signaling pathway. Low density lipoprotein receptor-related protein 1 (LRP1) is involved in the regulation of the inflammatory reaction induced by cytokines.
OBJECTIVE:To study the effect of knockdown of LRP1 on tumor necrosis factor α-induced inflammatory reaction.
METHODS: Primary cultured rat chondrocytes were transfected with lentivirus-mediated RNA interference to knockdown LRP1 gene. Three days after lentivirus transfection, chondrocytes were pretreated with Bay 11-7082 (10 μmol/L) for 30 minutes prior to the addition of tumor necrosis factor α (30 μg/L) for 30 minutes. Signaling protein and mRNA expressions in chondrocytes were detected by western blot assay and real-time PCR analysis, respectively. Chondrocytes were pretreated with or not Bay 11-7082 (10 μmol/L) 30 minutes prior to the addition of tumor necrosis factor α (30 μg/L) for 12 hours after starvation in DMEM for overnight, and the culture medium was colected for ELISA determination of matrix metaloproteinase 13 level.
RESULTS AND CONCLUSION:Tumor necrosis factor α receptor 1 expression was upregulated in chondrocytes after lentivirus-induced knockdown of LRP1. Increased expression of inducible nitric oxide synthase and activation of the nuclear factor kappa B signaling pathway were found after the addition of tumor necrosis factor α in shLRP1 group. Moreover, increased level of matrix metaloproteinase 13 was determined by ELISA. Taken together, knockdown of LRP1 up-regulates the expression of tumor necrosis factor α-induced inducible nitric oxide synthase and matrix metaloproteinase 13 through the activation of the nuclear factor kappa B signaling pathway.
