N-methyl-D-aspartate receptor blockers attenuate bleomycin-induced pulmonary fibrosis by inhibiting endogenous mesenchymal stem cells senescence

Pu Huang; Yan Zhou; Xiao-Hong Li; Yun-Na Zhang; Hai-Peng Cheng; Jia-Feng Fu; Wei Liu; Shaojie Yue; Zi-Qiang Luo

doi:10.21037/atm-22-2507

N-methyl-D-aspartate receptor blockers attenuate bleomycin-induced pulmonary fibrosis by inhibiting endogenous mesenchymal stem cells senescence

Ann Transl Med. 2022 Jun;10(11):642. doi: 10.21037/atm-22-2507.

Authors

Pu Huang^{1

2}, Yan Zhou¹, Xiao-Hong Li³, Yun-Na Zhang¹, Hai-Peng Cheng³, Jia-Feng Fu¹, Wei Liu⁴, Shaojie Yue⁵, Zi-Qiang Luo^{1

6}

Affiliations

¹ Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, China.
² Health Management Center, Changsha Central Hospital Affiliated to Nanhua University, Changsha, China.
³ Department of Pathology, The Second Xiangya Hospital, Central South University, Changsha, China.
⁴ Xiangya Nursing School, Central South University, Changsha, China.
⁵ Department of Pediatrics, Xiangya Hospital, Central South University, Changsha, China.
⁶ Hunan Key Laboratory of Organ Fibrosis, Central South University, Changsha, China.

Abstract

Background: A large number of our previous studies showed that endogenous glutamate and N-methyl-D-aspartate receptor (NMDAR) activation may be involved in various types of acute lung injury, airway inflammation, asthma, and pulmonary fibrosis. In animal models, the transplantation of exogenous bone marrow mesenchymal stem cells (BM-MSCs) is the most promising treatment for idiopathic pulmonary fibrosis. However, there are limited reports on the status of endogenous BM-MSCs in the process of bleomycin-induced pulmonary fibrosis in animals.

Methods: We constructed a mouse model of bleomycin-induced pulmonary fibrosis. In vitro, the senescence model of BM-MSCs was constructed with hydrogen peroxide and high concentration of N-methyl-D-aspartate (NDMA). The changes in aging-related indexes were detected by senescence associated beta-galactosidase (SA-β-gal) staining, western blot, flow cytometry and real time-PCR. The epithelial-mesenchymal transformation (EMT) changes of mouse lung epithelial cells (MLE-12) co-cultured with senescent BM-MSCs were detected by immunofluorescence and western blotting.

Results: We observed that endogenous BM-MSCs senescence occurs during bleomycin-induced pulmonary fibrosis in mice, and the model group had a higher expression level of the NMDAR subunit than the control group. We observed a significant increase in NMDAR subunit expression in a hydrogen peroxide-induced senescent cell model in vitro. BM-MSCs showed senescence-related phenotype and cell cycle arrest after high concentration of NMDA treatment. At the same time, the expression levels of the classic Wingless and int-1 (Wnt) pathway protein β-cantenin and downstream cyclin D1 also changed. In the co-culture of aged BM-MSCs and MLE-12 cells, EMT can be promoted in MLE-12 cells, and MK-801 can partially antagonize the occurrence of EMT. The NMDAR antagonist can partially prevent the above phenomenon.

Conclusions: High concentrations of NMDA can promote senescence of BM-MSCs. NMDAR blockers may inhibit endogenous BM-MSCs aging through the WNT signaling pathway, thereby reducing the effect of bleomycin-induced pulmonary fibrosis.

Keywords: Bone marrow mesenchymal stem cells (BM-MSCs); N-methyl-D-aspartate receptor (NMDAR); idiopathic pulmonary fibrosis (IPF); senescence.