Objective To investigate the
therapeutic effect of Bone marrow mesenchymal stem cells (BMSCs) on
dry eye mice, and to investigate the mechanism of TLR4/MYD88/NF-κB signaling pathway on
corneal injury repair in
dry eye mice. Methods To establish a hypertonic
dry eye cell model. Western blot for measureing the
protein expressions of caspase-1, IL-1β,NLRP3 and ASC,and Rt-qpcr for
mRNA expression. Flow cytometry for detecting the ROS content and apoptosis rate.
CCK-8 for detecting the proliferation activity of cells, and ELISA for the levels of
inflammation-related factors.The levels of
inflammation-related factors were detected by ELISA. The
dry eye mouse model of
benzalkonium chloride was established. Three clinical parameters used to evaluate ocular surface damage, namely tear secretion, tear film
rupture time and corneal
sodium fluorescein staining, were measured with
phenol cotton thread. Flow cytometry and TUNEL staining are both for he apoptosis rate. Western blot also for detecting the
protein expressions of TLR4, MYD88, NF-κB,
inflammation-related factors and apoptosis-related factors . The pathological changes were evaluated by HE and PAS staining. Results In vitro, BMSCs and inhibitors of TLR4, MYD88 and NF-κB showed decreased ROS content, decreased inflammatory factor
protein level, decreased apoptotic
protein level and increased
mRNA expression compared with NaCl group. BMSCS partially reversed cell apoptosis induced by NaCl and improved cell proliferation. In vivo, it reduces corneal epithelial defects, goblet cell loss and inflammatory
cytokine production, and increases tear production. In vitro, BMSC and inhibitors of TLR4, MYD88 and NF-κB could protect mice from apoptosis induced by hypertonic stress. In terms of mechanism, NACL-induced NLRP3
inflammasome formation, caspase-1 activation and IL-1β maturation can be inhibited. Conclusion BMSCs treatment can reduce ROS and
inflammation levels and alleviate
dry eye by inhibiting TLR4/MYD88/NF-κBsignaling pathway.