热休克蛋白90通过蛋白激酶B通路在局灶性脑缺血再灌注损伤中的作用机制研究

Role of HSP90 in focal cerebral ischemia/reperfusion injury through protein kinase B pathway

目的探讨热休克蛋白90(HSP90)在大鼠局灶性脑缺血再灌注损伤中的作用机制及丹参多酚酸盐干预的影响。方法采用线栓法制作SD大鼠脑缺血再灌注损伤模型。选用健康雄性SD大鼠75只随机分为对照组、模型组、抑制剂组、丹参多酚组及联合组,每组15只。对照组仅暴露左颈总动脉及颈内外动脉后缝合皮肤。模型组建立缺血再灌注模型,抑制剂组、丹参多酚组及联合组分别于建立缺血再灌注模型后给予抑制剂、丹参多酚酸盐及抑制剂联合丹参多酚酸盐。比较各组脑梗死体积,Western blot法测定HSP90、蛋白激酶B(Akt)、磷酸化Akt(p-Akt)变化。结果与模型组比较,对照组和丹参多酚组脑梗死体积明显减少,差异有统计学意义[0 mm3和(163.11±9.12)mm3 vs(268.92±14.36)mm3,P<0.05]。与联合组比较,抑制剂组脑梗死体积明显增加,差异有统计学意义[(237.13±11.11)mm3 vs(219.08±7.44)mm3,P<0.05]。与模型比较,对照组HSP90蛋白表达水平明显降低,丹参多酚组HSP90的蛋白表达水平明显升高(P<0.05)。与模型组比较,对照组和抑制剂组p-Akt蛋白表达明显降低,丹参多酚组p-Akt蛋白表达显著升高,差异具有统计学意义(P<0.05,P<0.01)。与丹参多酚组比较,联合组p-Akt蛋白表达水平明显降低,差异有统计学意义(P<0.05)。结论 HSP90可能通过Akt/p-Akt通路在脑缺血再灌注损伤中发挥神经保护作用。

Objective To study the role of HSP90 in and the effect of salvianolic polyphenol on focal cerebral ischemia/reperfusion（I/R）injury.Methods A rat cerebral I/R injury model was established by Longa occlusion.Seventy-five SD rats were divided into control group,model group,inhibitor group,salvianolic polyphenol treatment group,and I/R injury ＋inhibitor group（15 in each group）.After I/R injury model was established,the infarction size in different groups was measured amd expressions of HSP90,Akt and P-Akt protein were detected by Western blot.Results The infarction size was significantly smaller in control group and salvianolic polyphenol treatment group than in model group（P〈0.05）and was significantly larger in inhibitor group than in I/R injury＋inhibitor group（P〈0.05）.The expression level of HSP90 and p-Akt protein was significantly lower in model group than in control group and was significantly higher in salvianolic polyphenol treatment group than in model group（P〈0.05,P〈0.01）.The expression level of p-Akt protein was significantly lower in inhibitor group than in model group and in I/R injury＋inhibitor group than in salvianolic polyphenol treatment group（P〈0.05）.Conclusion HSP90 protects nerves against injury through the Akt/P-Akt pathway in cerebral I/R injury.