川芎挥发油对大鼠大脑皮层神经细胞体外存活及脑缺血再灌注损伤的影响

Effects of Aetherolea from Chuanxiong on Ischemical Reperfusion Injury in Rats and the Survival of Cerebral Cortex Neurons in vitro

目的考察川芎挥发油对大鼠大脑皮层神经细胞的体外存活及脑缺血再灌注损伤的保护作用。方法取出生1d内的乳鼠大脑皮层制成细胞悬液,接种于经多聚赖氨酸包被的培养板。于培养6 d后,加入不同浓度的川芎挥发油,继续培养1 d,采用MTT比色法测量其存活细胞的吸收值,同时行NSE检查。采用线栓法阻塞大脑中动脉(MCA)致大鼠局灶性脑缺血再灌注损伤模型,测定大鼠血清中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、一氧化氮合酶(NOS)含量。结果 NSE检查结果表明,培养6 d的存活细胞大部分均为神经元。MTT比色法结果表明,与对照组比较,川芎挥发油较低浓度均能明显增加存活细胞数(P<0.05)。且川芎挥发油能明显提高GSH-Px、NOS、SOD的活性,降低MDA的含量及大鼠脑梗塞比率(P<0.05或P<0.01)。结论川芎挥发油能促进体外培养的大脑皮层神经细胞存活,可通过增加脑缺血再灌注模型大鼠SOD、GSH-Px、NOS活性,降低MDA含量,降低大鼠脑梗塞比率等途径产生脑缺血保护作用。

Objective To explore the effect of aetherolea from Chuanxiong on ischemical reperfusion injury in rats and the survival of cerebral cortex neurons. Methods The cerebral cortex of neonate Sprague-Dawley rats was dissociated into cell suspension,then incubated in culture plates covered with polylysine.After cultured for 6 days,aetherolea from Chuanxiong added to the 96-well culture plates,continued to culture for 1 day.Then,the OD of living cells in each well was measured by MTT assay.Some of the 6-day culture cells were stained with neuronal specific nolase（NSE） antibody.The model of middle cerebral artery occlusion（MCAO） was established by suture embolism in rats,and the cerebral infarction proportion,SOD,MDA,GSH-Px and NOS in serum were established. Results Most of the living cells were cerebral neurons by NSE.Compared with control group,low concentrations of aetherolea from Chuanxiong could increase the viable count significantly in vitro（P0.01）,decrease cerebral infarction ratio,enhance the activities of SOD,GSH-Px and NOS,and decrease the content of MDA in rats（P0.01）. Conclusion Aetherolea from Chuanxiong can promote the live of cerebral cortex neurons in vitro,decrease cerebral infarction volume,enhance the activities of SOD,GSH-Px,NOS,decrease the content of MDA in rats,and alleviate the damages caused by ischemical reperfusion.