金雀异黄酮预适应对大鼠小脑浦肯野细胞缺血再灌注损伤的保护作用

Protective effect of Genistein preconditioning on ischemia-reperfusion injury of cerebellar Purkenje cells in rats

目的建立糖氧剥夺(OGD)的缺血再灌注模型,观察金雀异黄酮(GEN)预适应对小脑浦肯野细胞缺血再灌注损伤的保护作用。方法制备SD大鼠小脑脑片,人工脑脊液灌流培养。脑片分为正常对照组、缺血再灌注(I/R)组、GEN 200、100、50 mmol/L预适应组和尼莫地平组。GEN培养20 min,OGD 20 min,再灌注4 h。采用HE染色,观察GEN作用下小脑浦肯野细胞未受损伤的百分比,检测小脑组织中肌酸肌酶(CK)活力。结果与正常对照组比较,I/R能明显诱导小脑浦肯野细胞死亡(P<0.000 5)。与I/R组比较,GEN预适应(200和100 mmol/L)小脑浦肯野细胞存活率明显升高(P<0.005和P<0.01),小脑组织中CK活力明显下降(P<0.01和P<0.001)。与I/R组比较,应用钙通道阻断剂尼莫地平,小脑浦肯野细胞存活率明显升高(P<0.01),小脑组织中CK活力明显下降(P<0.01)。结论 GEN对小脑浦肯野细胞I/R损伤具有保护作用。

Objective To observe the protective effects of Genistein on ischemia-reperfusio/l （I/R） injury of cerebellar Purkenje cells. Methods Cerebellar brain slices from SD rats were incubated with oxygenated artificial cerebrospinal fluid. They were randomly divided into six groups： control I/R, GEN 200, 100, 50 μmol/L preconditioning and Nimodipine groups. Cerebellar slices were incubated with GEN for 20 rain before subjected to simulated ischemia （ oxygen-glucose deprivation） for 20 rain. After ischemia, they were recovered in oxygenated artificial cerebrospinal fluid for 4 h. Then they were fixed and stained by hematoxylin-eosin （HE） for morphologic examination to determine the percentage of undamaged Purkinje cells. The activity of CK in cerebellar brain slices was detected. Results Compared with control group, the survival rate of Purkinje cells was significantly lower after oxygen-glucose deprivation and reoxygenation （ P 〈 0. 000 5 ）. Compared with I/R group, the survival rate of Purkinje cells was significantly higher in GEN 200 and 100 mmol/L groups （P 〈 0. 005, P 〈 0.01 ）, and the activity of CK was significantly lower （P 〈0. 001, P 〈0. 01 ）. Compared with I/R group, the survival rate of Pnrkinje cells was significantly higher in slices incubated with Nimodipine （P 〈0. 01 ）, and the activity of CK was significantly lower （P 〈 0. 01 ）. Con- clusions GEN has protective effect on cerebellar Purkenje cells injured by I/R.