Edaravone

Edaravone (Mitsubishi Tanabe Pharma Corporation, Osaka, Japan) is a radical scavenger and used as a treatment drug for cerebral infarction^{11 (link)}. We examined whether Edaravone could inhibit skeletal muscle injury after the ischemia–reperfusion experiment. A rat model subjected to 4 h of ischemia and 24 h of reperfusion (n = 4) was created and samples of blood and skeletal muscle (gastrocnemius in both cases) were taken. In this protocol Edaravone (9.0 mg/kg) was administered into the peritoneal cavity at the beginning of the reperfusion and 12 h later according to the report by Yamamura et al.^{12 (link)}, although peritoneal cavity administration is an off-label use of Edaravone.
Additional MRI examinations with Edaravone were performed using the same protocol mentioned above (n = 6 each group) to evaluate whether Edaravone could inhibit the increase in 3-CP SI by eliminating ROS. In this experiment, we adopted a model of strong oxidative stress resulting from 4 h of ischemia and 24 h of reperfusion.

Edaravone was supplied from Mitsubishi Tanabe Pharma Corporation, Tokyo, Japan. For the randomized control study, we arbitrarily determined to analyze 10 animals per group, in the similar way to previous studies [11 (link)–18 (link)]. Each mouse was assigned confidentially in numerical order into one of 3 different doses of Edaravone groups (0 mg/kg, 1 mg/kg, and 10 mg/kg). This automatic procedure of randomization thoroughly eliminated the controller's bias, and it was difficult to conceive the dose of Edaravone for each mouse by its number. The researchers who participated in this study were strictly blind to the information of treatment. The wobbler mice initially developed shaking body from the age of 3 to 4 weeks, and were diagnosed as the symptomatic onset. Immediately after diagnosis, Edaravone or vehicle was administered by intraperitoneal injection daily for 4 weeks. Finally, participating affected mice were divided into three groups: the low dose Edaravone group (1 mg/kg, n = 10), the high dose Edaravone group (10 mg/kg, n = 10) and the vehicle group (n = 10). The treatment finished at the age of 7 to 8 weeks.

In humans, 30 mg of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one; Radicut^®) was intravenously infused. Notably, edaravone was provided by Mitsubishi Tanabe Pharma Corporation (Osaka, Japan). It was dissolved in a small volume of 1 M NaOH solution. The pH of the solution was adjusted to 7 using 1 M HCL in accordance with the relevant previous studies [16 (link),20 (link)]. The prepared edaravone was used as a solution. Further, the concentration was adjusted to 1.5, 3, or 10 mg/mL in 0.9% (physiologic) saline solution in accordance with the relevant previous studies [15 (link),21 (link),22 (link)]. Previous studies reported that 15, 30, and 100 mg/kg edaravone can improve ALS, radiation-induced oral mucositis, and cisplatin-induced renal injury, respectively [15 (link),21 (link),22 (link)]. Subsequently, 15, 30, or 100 mg/kg edaravone was injected intraperitoneally 30 min (min) before irradiation. In contrast, saline was injected in the control group.

Edaravone synthesized at Mitsubishi Tanabe Pharma Corporation (Osaka Japan) was added to the cells infected with adenoviruses expressing the WT and CTF TDP-43 at final concentrations of 1–200 μmol/L in the cell viability assay and 50 μmol/L in the sequence analysis assay. Edaravone was prepared in DMSO (100%), and the final concentration of DMSO was set at 0.1% in both vehicle and Edaravone treatments. Twenty-four hours later, the cells received a medium either containing or not containing 20 μmol/L ethacrynic acid (#SML1083; Sigma) and were further incubated for 24 h.