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目的研究全脑缺血再灌注给予δ阿片受体激动剂DADLE前后大鼠脑组织Bcl-2、Caspase-3的表达情况,探讨DADLE在全脑缺血再灌注损伤中对Bcl-2、Caspase-3的影响及与脑保护作用的关系。方法健康雌性SD大鼠50只(180-220g)随机分为5组:假手术组(Sham,n=10):只暴露血管而不夹闭;缺血再灌注组(I/R,n=10):采用改良的二血管阻断加低血压法建立全脑缺血再灌注模型,缺血15min后给予再灌注;DADLE处理组分为2mg/kg组(A,n=10)、DADLE 3mg/kg组(B,n=10)、DADLE 5mg/kg组(C,n=10):在I/R组的基础上于再灌注前经颈外静脉注射DADLE。各组动物再灌注120min后处死,开颅取大脑组织,采用western-bolt技术检测大鼠脑组织Bcl-2、Caspase-3的表达状况。结果 western-bolt检测结果显示,sham组Bcl-2表达水平显著低于其他各组(P<0.05),且Sham组仅少量Caspase-3表达。DADLE处理组与I/R组相比,Bcl-2表达水平上调(P<0.05)而Caspase-3表达水平下调(P<0.05)。各给药组之间Bcl-2为B组与C组的表达显著高于A组(P<0.05),B组与C组间差异无统计学意义(P>0.05);Caspase-3表达水平为B组与C组的表达显著低于A组(P<0.05),而B组与C组间差异无统计学意义(P>0.05)。结论药物DADLE可能通过上调Bcl-2的表达并抑制Caspase-3的表达,抑制神经细胞的凋亡,减低全脑缺血再灌注损伤,从而对大脑起保护作用。
Objective To investigate the expression of Bcl-2 and Caspase-3 in rat brain before and after global cerebral ischemia-reperfusion with D opioid receptor agonist DADLE. To investigate the effect of DADLE on the expression of Bcl-2, Caspase- 3 and its relationship with brain protective effect. Methods Fifty healthy female Sprague Dawley rats (180-220g) were randomly divided into 5 groups: sham operation group (Sham, n = 10): only vessels were exposed but not occluded; ischemia / reperfusion group 10): The model of global cerebral ischemia-reperfusion was established by modified two-vessel occlusion plus hypotension. The rats were reperfused 15 min after ischemia. The rats in DADLE group were treated with 2 mg / kg (A, n = 10) (B, n = 10), DADLE 5 mg / kg (C, n = 10): DADLE was injected via the external jugular vein on the basis of the I / R group before reperfusion. The animals in each group were sacrificed 120 min after reperfusion. Brain tissue was taken out by craniotomy. Western-bolt technique was used to detect the expression of Bcl-2 and Caspase-3 in rat brain. Results The results of western-bolt showed that the expression of Bcl-2 in sham group was significantly lower than that in other groups (P <0.05), and only a small amount of Caspase-3 was expressed in Sham group. Compared with I / R group, Bcl-2 expression increased (P <0.05) and Caspase-3 expression decreased in DADLE treatment group (P <0.05). The expression of Bcl-2 in B group and C group was significantly higher than that in A group (P <0.05), and there was no significant difference between B group and C group (P> 0.05). The expression of Caspase-3 There was no significant difference between group B and group C (P> 0.05). Conclusion DADLE may protect brain by up-regulating the expression of Bcl-2, inhibiting the expression of Caspase-3, inhibiting the apoptosis of nerve cells and decreasing the global ischemia-reperfusion injury.