The tissue micromilieu in disorders (
inflammation,
ischemia,
tumor) often shows pronounced
metabolic acidosis that may alter signaling and transcriptional activity in resident cells which can be of special importance for omnipresent fibroblasts. In the present study we investigated the impact of
metabolic acidosis on rat fibroblasts with special emphasis on their role in
inflammation by regulation of TNF-α, MCP-1, COX-2 and iNOS expression and the signaling pathways involved. Extracellular
acidosis led to an enhanced expression of TNF-α, COX-2 and iNOS in parallel to an activation of p38 and ERK1/2
kinases that was not observed by sole intracellular
acidosis. Accordingly, the
protein amounts of TNF-α and COX-2 as well as the production of
nitrate and
nitrite were elevated.
Acidosis-induced expression of COX-2 and iNOS depended on p38
kinase, but not on ERK1/2. In contrast
acidosis-induced TNF-α expression was independent of both
kinases. Although GPR4, GPR68 and GPR132 are expressed in fibroblasts, the involvement of these potential candidate pH sensors could be ruled out since no
acidosis-induced elevation in intracellular cAMP or free
calcium content was observed. Furthermore our data show that MAPK activation by an acidic micromilieu depends on Ser/Thr
phosphatase activity, but not on the production of
reactive oxygen species and is sensitive to cAMP antagonism by
Rp-cAMPS. In conclusion, our results show that an acidic microenvironment induces a differential transcriptional program of pathological relevant genes in fibroblasts via the cAMP-
phosphatase-MAPK pathway and thereby generates a parainflammatory situation that can result in tissue remodeling.