MG53 attenuates nitrogen mustard‐induced acute lung injury

Abstract

Sulphur mustard (SM) and nitrogen mustard (NM) are alkylating agents known to cause severe damage to organs including the skin, eyes, lungs and nervous system, among which pulmonary toxicity is the major cause of death. 1 Pulmonary toxicity involves complicated cellular events, including DNA damage, oxidative stress, acute injury and inflammation. [2][3][4] In those who survive vesicant exposure, progressive inflammation often leads to pulmonary fibrosis and prolonged respiratory dysfunction. [5][6][7][8] Pulmonary exposure to SM/NM induces alkylating injuries with an acute and a chronic injury phase. The acute lung injury develops due to oxidative stress, lipid peroxidation and liberation of inflammatory mediators. [9][10][11] The prolonged injury is mediated by chronic inflammation and fibrotic remodelling in the upper and small airways. 12,13 Despite many studies investigating the mechanism of mustard-induced lung injury, current therapies to treat mustard poisoning are mostly palliative. [4][5][6][7][8][14][15][16][17][18][19] Therapeutic means that mitigate acute oxidative stress, harness inflammation and restore cell membrane integrity could potentially alleviate the deleterious impact of vesicant and other environmental insults in the lung.MG53 is a member of the TRIM protein (TRIM72), which functions as an essential component of plasma membrane repair. 20,21 MG53 knockout mice (mg53 −/− ) develop pulmonary pathology due

MG53 Mitigates Nitrogen Mustard-Induced Skin Injury

“…These findings are consistent with our previous study that reveals a role for oxidative stress in the control of MG53 s membrane repair function [54,55]. Similar findings were also observed with lung epithelial and endothelial cells [46]. Thus, NM-induced tissue injury and oxidative stress can all be mitigated by rhMG53, laying the foundation for the use of exogenous rhMG53 to boost the defense mechanism against vesicant-induced tissue injury.…”

“…Cell apoptosis was investigated by dual staining with Alexa Fluor 488-Annexin V and Propidium Iodide (PI) (Invitrogen Cat# V13241) following the manufacture protocol and analyzed as described previously [46,47]. Briefly, HFSC cells were seeded in 6-well plates and cultured for 24 h, then incubated with 50 µM NM or BSA for 4 h and incubated with vehicle or rhMG53 (10 µg/mL) for another 20 h. Cells were detached by 0.25% Trypsin-EDTA solution.…”

“…Cellular reactive oxygen species (ROS) production was measured using a Cellular ROS Detection Assay Kit (Abcam Cat#ab113851) according to manufacturer instructions and analyzed as described previously [46,47]. Briefly, HFSC cells were seeded in 6 well plates and cultured for 24 h, then incubated with 50 µM NM or BSA for 4 h, and incubated vehicle or rhMG53 (10 µg/mL) cultured for another 20 h. Cells were washed with PBS and re-suspended with 100 µL assay buffer including 1× ROS Deep Red Dye and incubated in 5% CO 2 , 37 • C for 30 min.…”