Neuroprotective effects of acacetin in oxygen-glucose deprivation in SH-SY5Y cells and cerebral ischena/reperfusion in mice

Abstract

Objective To investigate the neuroprotective effects of acacetin in oxygen-glucose deprivation in SH-SYSY cells and cerebral ischemia/reperfusion in mice. Methods Neuroblastoma cells （SH-SYSY） were cultured, and they were divided into the normal group, oxygen-glucose deprivation （OGD） group, and the groups of 1, 5 and 10 μmol/L acacetin. After 24-h reoxygenation, the MTT method was used to assay the cell viability and lactate dehydrogenase （LDH） assay was used to measure the LDH release rate. A total of 60 C57 mice were randomly divided into 5 groups： the sham operation group, the ischemia-reperfusion group and the groups of low moderate- and high-dose acacetin （n = 12 in each group）. A model of middle cerebral artery occlusion （MCAO） was induced by the intraluminal suture method. During the ischemia-reperfusion period, acacetin was injected intraperitoneally （6. 25, 12. 5, and 25 mg/kg, respectively ）. The neurological function score was evaluated at 24 hours after reperfusion. Cerebral infarct volume was determined by using 2, 3,5-triphenyl tetrazolium chloride staining. Results The in vitro experiments showed that the SH-SY5Y cell survival rates in the groups of 1 μmol/L, 5 txmol/L and 10 μmol/L acacetin were 90. 34% ± 6. 87% （P = 0. 000）, 85. 47% 2. 24% （P =0. 001） and 81.79% ± 1.77% （P =0. 008）, respectively, they were significantly higher than that in the OGD group （70. 62% ± 8. 89%）. The LDH release rates in the groups of 1 izmol/L （P = 0. 021 ）, 5 txmol/L （P =0. 011） and 10 txmol/L （P = 0. 027） acacetin were 159. 11% ± 13. 11%, 155. 12% ± 24. 72% and 160. 92% ±7. 83%, respectively, they were significantly lower than that in the OGD group （180. 35% ± 10. 60%）. The in vivo experiments showed that the neurological function score in the group of high-dose acacetin was lower than that in the ischemia-reperfusion group （1.67 ±0. 85 vs. 2. 50 ±0. 55; P =0. 018）; the infarct volumes in the groups of low-, moderate-, and high-dose acacetin were 24. 14 ± 7. 1 mm3, 17. 18 ± 3.19 mm3, and 12. 86 ± 1.88 mm3, respectively, they were significantly smaller than that in the ischemia- reperfusion group （48.81 ±9. 48 mm3） （all P =0. 000）. Conclusions Both in vitro and in vivo experinaents have shown that acacetin has the neuroprotective effect. Key words: Acacetin; Cell Line; Cell Survival; Oxyn; Glucose; L-Lactate Dehydroase; Cells,Cultured; Brain Ischemia; Disease Models, Animal; Mice