Free radicals and oxidative stress play key roles in cerebral ischemic pathogenesis and represent pharmacological targets for treatment. Edaravone (Edv), one of antioxidant agents that have been used in acute ischemic stroke in both clinical settings and animal experiments, exerts neuroprotective effect on ischemic injured brains. This review is aimed to elaborate the latest molecular mechanisms of the neuroprotection of Edv on cerebral ischemia and provide reasonable evidence in its clinical application. It is found that Edv has neuroprotective influence on cerebral ischemia, which is closely related to the facets of scavenging reactive oxygen species (ROS), hydroxyl radical (ċOH) and reactive nitrogen species (RNS). And it is a good antioxidant agent that can be safely used in the treatment of cerebral ischemia and chronic neurodegenerative disorders as well as other ischemia/reperfusion (I/R)-related diseases. The combination of Edv with thrombolytic therapy also can be applied in clinical settings and will be greatly beneficial to patients with stroke.

The present study aimed to establish an effective method for the in vitro culture of guinea pig airway smooth muscle (ASM) cells, and also investigate the suppressive effect of mabuterol hydrochloride (Mab) on the increased level of intracellular Ca2+ in ASM cells induced with acetylcholine (Ach). Two different methods, i.e. with or without collagenase to pretreat tracheal tissues, were applied to the manufacture of ASM cells. Cell viability was determined with the 3-(4,5-dimethylthinazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Immunocytochemistry and immunofluorescence were used for the identification of ASM cells. Different concentration levels (10−3, 10−4, 10−5, 10−6 and 10−7 mmol/l) of Mab were administered 5 min before Ach (10−4 M) treatment, respectively. The Ca2+ fluorescent probe, Fura-2/AM or Fluo-3/AM were applied to the inspection of Ca2+ fluorescent intensity with Varioskan Flash, immunocytometry systems and an inverted system microscope, respectively. The results showed that the fresh method, in which isolated tracheal tissues were previously treated with collagenase for 20 min, was more advantageous for the preparation of guinea pig ASM cells compared to when the enzyme was not used. The time for the ASM cells to initially migrate out of the ‘tissue blocks’ and the culture having to be generated due to the thick cell density was significantly less. On identification with immunocytochemistry or immunofluorescent staining, >95% of the cells were ASM cells. Mab (10−3−10−7 mmol/l) significantly suppressed the elevation of intracellular Ca2+ induced by Ach in a concentration-dependent manner. The inhibitory rates of intracellular Ca2+ by different concentrations of Mab, from low to high, were 14.93, 24.73, 40.06, 48.54 and 57.13%, respectively, when Varioskan Flash was used for determination. In conclusion, this novel method has a shorter harvesting period for ASM cells. Mab can suppress the increasing level of intracellular Ca2+ induced by Ach in guinea pig ASM cells. Further investigation into the precise mechanisms of action is required.