Neuroprotective effects of an alkaloid-free ethyl acetate extract from the root of Sophora flavescens Ait. against focal cerebral ischemia in rats.
Large amounts of brain nitric oxide are produced over several hours after a stroke. This probably causes DNA strand nicks, nitration of cytosolic components of neurons, and ultimately neuronal death. Oxymatrine and matrine are two major alkaloids of the Chinese herb Sophora flavescens Ait. (Leguminosae); they have been demonstrated to inhibit liver injury during warm ischemia and reperfusion and to induce apoptosis, respectively, in vivo and in vitro. However, the neuroprotective efficacy of the EtOAc extract of S. flavescens (ESF) without the alkaloids has not been explored. This study investigated the inhibitory efficacy of ESF, which contain two major flavonoids kurarinone (45.5%) and sophoraflavone G (14.7%), in focal cerebral ischemia. Focal cerebral ischemia was induced using the middle cerebral artery occlusion (MCAO) method. After 1.5h of MCAO and 24h of reperfusion, the extent of neurological deficits and the infarct volume were measured in Sprague-Dawley rats. Compared with carnosine (50mg/kg), as positive control ESF (20mg/kg) significantly reduced infarct volume and neurological deficits. Treatment of human SH-SY5Y cells with sodium nitroprusside (SNP), a nitric oxide donor, decreased cell viability by causing apoptosis-like cell death. ESF significantly inhibited caspase-3-like enzyme activity and DNA fragmentation. The level of active caspase-3 was maximal 6h after SNP treatment. However, active caspase-3 and apoptosis were dose-dependently inhibited by ESF treatment. Flow cytometry analysis showed that ESF significantly inhibited cell apoptosis (p<0.05) and reduced the apoptotic index by 79.9% (p<0.01). These results indicate that ESF is neuroprotective in focal cerebral ischemia and the flavonoids in ESF might be responsible for its neuroprotective activity in rats, alone or in part.
Natural Products Research Institute, College of Pharmacy, Seoul National University, Gwanak-gu, Seoul 151-742, Republic of Korea.
S J
SJ
Park
K W
KW
Nam
H J
HJ
Lee
E Y
EY
Cho
U
U
Koo
W
W
Mar
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D053938:DNA Fragmentation / Q000187:drug effects; D006801:Humans; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy*; D008297:Male; D018696:Neuroprotective Agents / Q000302:isolation & purification* / Q000494:pharmacology / Q000627:therapeutic use; D009599:Nitroprusside; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D018517:Plant Roots / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D029909:Sophora / Q000737:chemistry*
D018696:Neuroprotective Agents; D010936:Plant Extracts; D009599:Nitroprusside; D053148:Caspase 3