engleski

Mechanisms of extracellular Hsp70 induced inflammatory effects in 16HBE cells

Extracellular heat shock protein (eHsp) 70 is a damage-associated molecular pattern that could participate in inflammatory response in bronchial epithelial cells. We hypothesized that the inflammatory response to eHsp70 as well as its effects on cell survival are mediated via mitogen-activated protein kinase (MAPK) and NF-κB signalling pathways in 16HBE bronchial epithelial cell line. We aimed to investigate these mechanisms using the specific NF-κB and MAPK inhibitors. 16HBE cells were pre-treated with specific inhibitors (BAY11-7082 for NF-κB, PD98059 for ERK, SP600125 for JNK, and SB202190 for p38) for 1 h before adding recombinant human (rh) Hsp70 (3 μg/ml) for 24 h. IL-1α, IL-6 and TNF-α concentrations were determined in cell supernatants using ELISA method, while cell viability was assessed by MTS assay. IL-1α secretion was decreased after pre-treatment with ERK, JNK and p38 inhibitors compared to rhHsp70 alone. IL-6 and TNF-α secretion was decreased with all inhibitors. Inhibition of NF-κB and p38 pathways almost completely attenuated secretion of TNF-α and IL-6. rhHsp70 alone did not affect cell viability ; however, inhibition of JNK and p38 pathways significantly lowered it in 16HBE cells. In conclusion, the results support our hypothesis that the NF-κB signalling is an important factor in inflammation, while MAPK signalling pathways are involved in both inflammatory processes and cell survival in 16HBE cells. This could have a potential role in new therapeutic drug discoveries.

16HBE cells ; extracellular Hsp70 ; MAPK, NF-kappaB ; cytokines

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