缺血后处理联合水蛭注射液后处理对脑缺血损伤大鼠磷酸化糖原合成酶激酶3β表达的影响

Effect of ischemic postconditioning combined with leech injection postconditioning on the expression of phosphorylated glyeogen synthase kinase-3β of ischemiareperfusion injury in rats

目的:观察缺血后处理联合水蛭注射液后处理对脑缺血损伤大鼠磷酸化糖原合成酶激酶3β(p-GSK3β)mRNA及其蛋白表达的影响。方法:SD大鼠40只,随机分为假手术组(SO组)、缺血再灌注对照组(MCAO组)、缺血后处理组(IPOC组)、缺血后处理+水蛭注射液后处理组(P-L组)4组。采用线栓法制备大鼠MCAO模型及IPOC模型,应用实时荧光定量PCR和Western blot技术检测p-GSK3βmRNA及其蛋白的表达变化。结果:SO组有少量p-GSK3βmRNA及其蛋白的表达;MCAO组大鼠脑缺血再灌注24h缺血半暗带p-GSK3β表达较SO组明显增高(P<0.01,P<0.05);IPOC组二者的表达水平较MCAO组明显升高(P<0.01,P<0.05);P-L组能较IPOC组进一步升高其表达(P<0.05)。结论:脑缺血后处理可能通过诱导p-GSK3β的表达发挥其神经保护作用,水蛭注射液后处理可进一步促进其表达而发挥神经保护作用。

Objective： To observe the effect of leech injection postconditioning combined with ischemic postconditioning on the mRNA and protein expression of phosphorylated glyeogen synthase kinase-3β （p-GSK3β） of ischemia-reperfusion injury in rats. Methods： 40 male SD rats were randomly divided into four groups： sham operation group （SO）, middle cerebral artery occlusion （MCAO） group, ischemic postconditioning group （IPOC） and leech injection postconditioning group （P-L）. The suture- occluded method was used to make the MCAO and IPOC model of rats. The real time PCR and western blot were used to test the mRNA and protein expression of p-GSK3β. Results： The mRNA and protein expression of p-GSK3β were both increased at 24h following reperfusion in MCAO, compared with SO group （P〈0.05）. Compared with MCAO group, the IPOC could increase the mRNA and protein expression of p-GSK3β （P〈0.05）. But the mRNA and protein expression of p-GSK3β in the P-L group were both higher than that of IPOC group. Conclusion： IPOC could protect neurons by adjusting the expression of p-GSK3β, and leech injection postconditioning could promote the expression induced by IPOC.