NMDA receptor activation inhibits the protective effect of BM‑MSCs on bleomycin‑induced lung epithelial cell damage by inhibiting ERK signaling and the paracrine factor HGF.

Endoplasmic reticulum (ER) stress in alveolar epithelial cells (AECs) is associated with the pathogenesis of pulmonary fibrosis. Bone marrow‑derived mesenchymal stromal cells (BM‑MSCs) can exert protective effects on ER‑stressed AECs via paracrine signaling. In the present study, mouse lung epithelial (MLE)‑12 cells were directly stimulated with various concentrations of bleomycin (BLM). MLE‑12 cell apoptosis was detected by flow cytometry, and Ki67 expression was detected by immunofluorescence to reflect cell proliferation. The results revealed that BLM increased the protein expression levels of X‑box binding protein 1 and immunoglobulin heavy chain‑binding protein, thus inducing ER stress, and caused cell dysfunction by inhibiting proliferation and promoting apoptosis. In addition, MSC‑derived conditioned medium (MSC‑CM) protected MLE‑12 cells from BLM‑induced injury, by reducing ER stress, promoting cell proliferation and inhibiting cell apoptosis. Our previous studies reported that N‑methyl‑D‑aspartate (NMDA) receptor activation partially inhibits the antifibrotic effect of BM‑MSCs on BLM‑induced pulmonary fibrosis through downregulating the paracrine factor hepatocyte growth factor (HGF). In the present study, the synthesis and secretion of HGF were detected by western blotting and ELISA, respectively. Results further demonstrated that NMDA inhibited the synthesis and secretion of HGF in BM‑MSCs, and NMDA‑preconditioned MSC‑CM had no protective effects on BLM‑induced injury in MLE‑12 cells. In addition, activation of the NMDA receptor decreased the phosphorylation levels of extracellular signal‑regulated kinase (ERK)1/2 in BM‑MSCs. Using Honokiol and FR180204, the activator and inhibitor of ERK1/2, respectively, it was then revealed that Honokiol partially eliminated the decrease in HGF expression, whereas FR180204 further promoted the reduction in HGF caused by NMDA. Collectively, these findings suggested that NMDA receptor activation may downregulate HGF by inhibiting ERK signaling in BM‑MSCs, thus weakening their protective effects on BLM‑induced lung epithelial cell damage.