Ninety [[C57BL/6 mice]] were randomized into [[sham]]-operated group, [[middle cerebral artery occlusion]] (MCAO) model group, and low-, medium and highdose (10, 20, and 40 mg/kg, respectively) FB groups. The expression levels of [[MDA]], [[ROS]], [[PCO]], [[8-OHdG]], [[SOD]], [[GSTα4]], [[CAT]] and [[GPx]] in the[[ brain tissue]] of the [[mice]] were detected using commercial kits, and those of [[AMPK]], [[P-AMPK]],[[ DAF-16]], [[FOXO3]] and [[P-FOXO3]] were detected with [[Western blotting]]. Compound C (CC), an AMPK inhibitor, was used to verify the role of the [[AMPK]] pathway in mediating the therapeutic effect of FB. In another 36 C57BL/6 mice randomized into 4 sham-operated group, MCAO model group, FB (40 mg/kg) treatment group, FB+CC (10 mg/kg) treatment group, TTC staining was used to examine the volume of cerebral infarcts, and the levels of ROS and SOD in the brain were detected; the changes in the protein expressions of AMPK, P-AMPK, DAF-16, FOXO3 and P-FOXO3 in the brain tissue were detected using Western blotting.

In mice with cerebral IR injury, treatment with FB significantly reduced the levels of ROS, MDA, PCO and 8-OHdG, increased the activities of antioxidant enzymes SOD, GSTα4, CAT and GPx, and enhanced phosphorylation of AMPK and FOXO3 and DAF-16 protein expression in the brain tissue (P < 0.01). Compared with FB treatment alone, the combined treatment with FB and CC significantly reduced phosphorylation of AMPK and FOXO3, lowered expression of DAF-16 and SOD activity, and increased cerebral infarction volume and ROS level in the brain tissue of the mice (P < 0.01).

FB inhibits [[oxidative stress injury]] caused by cerebral I/R in mice possibly by enhancing AMPK phosphorylation, promoting the downstream DAF-16 protein expression and FOXO3 phosphorylation, increasing the expression of antioxidant enzymes, and reducing [[ROS]] level in the brain tissue
((Chen X, Wang K, Chu D, Zhu Y, Zhang W, Cao H, Xie W, Lu C, Li X. [Forsythiaside B inhibits cerebral ischemia/reperfusion-induced oxidative stress injury in mice via the AMPK/DAF-16/FOXO3 pathway]. Nan Fang Yi Ke Da Xue Xue Bao. 2023 Feb 20;43(2):199-205. Chinese. doi: 10.12122/j.issn.1673-4254.2023.02.06. PMID: 36946038.)).