大鼠局灶性脑梗死后半胱氨酸天冬氨酸蛋白酶3,9的表达及白花丹参叶的干预效应

Expression of caspase-3,9 and the intervention effects of salvia miltiorrhiza beg.f.alba in rats with focal cerebral infarction

目的:观察大鼠局灶性脑梗死后促进细胞凋亡的半胱氨酸天冬氨酸蛋白酶3,9表达的变化规律及白花丹参叶的保护作用。方法:实验于2004-03/06在山东省泰山医学院微循环重点实验室完成。取50只Wistar大鼠,随机分为局灶性脑梗死组和白花丹参叶预处理组两组各25只,每组又按光照后时间分为30min,12,24,48,72h等5个亚组,每亚组5只。造模前,白花丹参叶预处理组大鼠给予20g/kg白花丹参叶水提物灌服,1次/d,连续7d,然后两组同时采用光化学法制作大鼠局灶性脑梗死的模型。利用免疫组化染色技术观察局灶性脑梗死后不同时间点大鼠脑半胱氨酸天冬氨酸蛋白酶3,9表达的变化,并观察大鼠一般情况。结果:50只大鼠全部进入结果分析。①局灶性脑梗死组大鼠梗死后30min即有半胱氨酸天冬氨酸蛋白酶3,9阳性细胞出现,呈半球状向四周放射样扩展,随梗死时间的延长逐渐增多,二者均在48h表达至高峰,但半胱氨酸天冬氨酸蛋白酶3表达更显著(P<0.05)。阳性细胞局限于梗死区周围。②预防性应用白花丹参叶水提物后,大鼠出现明显的嗜睡现象,受损皮质表面积红染区、蓝染区明显减小。各时点半胱氨酸天冬氨酸蛋白酶3,9的阳性反应均明显减少,较单纯梗死组有显著性差异(P<0.05)。结论:局灶性脑梗死后可诱导半胱氨酸天冬氨酸蛋白酶3,9的表达,白花丹参对脑梗死具有保护作用,其分子机制可能为抑制了半胱氨酸天冬氨酸蛋白酶3,9的表达。

AIM：To observe the change rules accelerates the apoptosis in rats with effects of salvia miltiorrhiza beg.f.alba. of the expression of caspase-3, 9 that focal cerebral infarction and the protective METHODS：The experiment was perfo,＇med at the Key l.aberatory of Microcirculation, Medical College of Taishan Medical University in Shandong Province between March and June 2004.Fifty wistar rats were selected and randomized into 2 groups：focal cerebral infarction group and salvia mihiorrhiza beg.f.alba pretreatment group with 25 in each group.Every group was divided into 5 subgroups： 30 minutes, 12, 24, 48, and 72 hours sub group according to the time after shining with 5 rats in the each subgroup.Before building model,the rats in the salvia mihiorrhiza beg.f.alba pretreatment group were treated with 20 g/kg aqueous extract of salvia mihiorrhiza beg.f.alba taking perfusing, once a day for 7 days, and then the rats in the two groups were made into focal cerebral infarction models with photochemical method at the same time. The changes of the expression of caspase-3, 9 at different time point in rats with focal cerebral infarction were observed with immunohistochemical staining technique, and the common status of rats was observed. RESULTS：Totally 50 rats were involved in the result analysis. ① There were positive cells of caspase-3,9 appeared at 30 minutes after infracting in the focal cerebral infarction group,showing hemisphere shaped and expanding around in a radial fashion,and progressively increased with time.Caspase-3,9- immunoreactivities were all increased to the maximum at the 48^th hour. But the expression of the former was more significant than the latter （P 〈 0.05）. The positive cells were limited to around infarcted region. ② There was significant drowsiness phenomenon appeared after the application of salvia mihiorrhiza beg.f.alba extract to prevent. The area of red or blue in injured cerebral cortex was decreased obviously. The positive reaction of caspase-3, 9 at each time point was reduced, and had markedly difference compared with the simple infarction group （ P 〈 0.05 ）. CONCLUSION：The focal cerebral infarction can induce the expression of caspase-3,9.The salvia miltiorrhiza beg.f.alba has protective effects on cerebra infarction, and its molecule mechanism may inhibit the expression of caspase-3, 9.