Protection of puerarin on neurons injured by mimic ischemia reperfusion in vitro
- VernacularTitle:体外模拟脑缺血再灌注诱导神经元损伤后葛根素的保护作用
- Author:
Juan CHEN
;
Jie ZHOU
;
Youmei FENG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2005;9(33):190-192
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Puerarin functions to relieve the injury caused by ischemia reperfusion, improve the microcirculation, and inhibit the agglutination of platelet. But it is not clear yet that whether its protection on neurons relates to the apoptosis of cells.OBJECTIVE: To observe the protection of puerarin on neurons injured by mimic ischemia reperfusion in vitro.DESIGN: A random control study. SETTING: Laboratory of Biochemistry and Moleculobiology Department of Preclinical Medicine College of Tongji Medical College of Huazhong University of Science and Technology.PARTICIPANTS: The experiment was done on March 11, 2002. The cultured N2a cells of rat neuromablast were observed.METHODS: Ninety minutes after the cultured N2a cells were put into the 37 ℃ incubator with 0.05 CO2 and 0.95 N2(v/v), puerarin 0.5 mmol/L was added for cuiture for 24 hours. The methyl thiazolyl tetrazolium (MTT)method was used to observe the surviving ability of cells. The Annexin-V staining was adopted to exam the severity of apoptosis in the early stage and the supernate was collected to analyze the activity of lactate dehydrogenase (LDH) to reflect the permeability of cytomembrane. The immunoblotting method was applied to analyze the expression of caspase-3, at the same time, its activity was measured.MAIN OUTCOME MEASURES: The severity of cell apoptosis, the activity of LDH, the expression and activity of caspase-3.RESULTS: Puerarin could improve significantly the surviving rate of N2a cells 24 hours after reperfusion, decrease remarkably the activity of LDH in the culture fluid, lessen obviously the N2a apoptosis (P ＜ 0.01), at the same time, reduce tremendously the activity and expression of caspase-3 induced by ischemia reperfusion (P ＜ 0.01).CONCLUSION: Puerarin can protect nerves, inhibit obviously the N2a apoptosis induced by ischemia reperfusion. The mechanism is that it inhibits greatly the expression and activity of caspase-3.
