Radioprotective effects and mechanism of HL-003 on radiation-induced salivary gland damage in mice

Abstract

Ionizing radiation (IR) can cause damage to the structure and function of salivary glands. Our research group independently synthesized the ROS scavenger, HL-003. The aim of this study was to explore the protective effects and underlying mechanisms of HL-003 on radiation-induced salivary gland injury. Salivary flow rate measurement, H&E staining, immunohistochemistry, FRAP, TUNEL, and western blotting were used to evaluate the radioprotective effect on salivary glands. The results showed that HL-003 protected the salivary secretion function by protecting the AQP-5 protein, on the salivary epithelial cell membrane, from IR damage. HL-003 reduced oxidative stress in the salivary gland by regulating the expression of ROS-related proteins NOX4, SOD2, and 8-OHdG. Furthermore, HL-003 downregulated the expression of p-p53, Bax, caspase 3, and caspase 9, and upregulated the expression of Bcl-2, suggesting that it could inhibit the activation of p53 to reduce cell apoptosis. In conclusion, HL-003 is an effective radioprotector that prevents damage of the radiation-induced salivary gland.

Amelioration of gamma irradiation‐induced salivary gland damage in mice using melatonin

“…The salivary gland function was evaluated by measuring SFR at different time points as described previously 16 …”

“…Salivary gland proteins were extracted as described previously 16 . We used 12% or 15% SDS‐PAGE gels to isolate proteins and transferred the proteins to polyvinylidene difluoride membranes.…”

“…Salivary gland proteins were extracted as described previously. 16 We used 12% or 15% SDS-PAGE gels to isolate proteins and transferred the proteins to polyvinylidene difluoride membranes. After blocking with 10% skim milk for 2 h at room temperature, the membranes were incubated with primary antibodies RPL11 (16277-1-AP; Proteintech), RPL6 (15387-1-AP; Proteintech), RPS12 (16490-1-AP; Proteintech), RPS20 (15692-1-AP; Proteintech), RPL17 (14121-1-AP; Proteintech), RPS27A (14946-1-AP; Proteintech), RPL36 (15145-1-AP; Proteintech), RPSA (14533-1-AP; Proteintech), RPL18A (14653-1-AP; Proteintech), RPL28 (16649-1-AP; Proteintech), Prkag1 (10290-1-AP; Proteintech), RPL35 (A17632; ABclonal), RPL13 (ab134961; Abcam), RPL14 (ab181200; Abcam), RPS16 (ab177951; Abcam), IRS2 (ab134101; Abcam), AP2S1 (ab128950; Abcam), RPS15 (ab157193; Abcam), RPL30 (ab170930; Abcam), MAPK13 (ab31828; Abcam).…”