Knockdown of low density lipoprotein receptor-related protein 1 in chondrocytes

doi:10.3969/j.issn.2095-4344.2016.15.006

Abstract

Abstract:

BACKGROUND: Tumor necrosis factor α, as a pathogenic factor, induces the inflammatory reaction mainly via 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 collected for ELISA determination of matrix metalloproteinase 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 metalloproteinase 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 metalloproteinase 13 through the activation of the nuclear factor kappa B signaling pathway.
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Chondrocytes, Tumor Necrosis Factor-alpha, NF-kappa B, Tissue Engineering

Yang Er-ping, Peng Fei, Liang Jie, Du Yuan-li. Knockdown of low density lipoprotein receptor-related protein 1 in chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(15): 2171-2177.

Figures/Tables 4

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