目的 分析远隔缺血后处理对新生大鼠缺氧缺血性脑病(HIE)脑组织钙敏感受体(CaSR)的表达和Akt/Pi3k通路的影响,为新生儿HIE的治疗提供理论依据。方法 7日龄新生Wistar大鼠120只随机分为假手术组(S组)、缺氧缺血性脑病组(HIE组)、远隔缺血后处理5 min组(R1组)、远隔缺血后处理10 min组(R2组)。观察各组大鼠行为表现并称其体重,HE染色观察细胞形态学变化,电镜观察细胞超微结构,免疫组化检测脑组织CaSR、Bax、Bcl-2、p-Akt、Akt蛋白的表达,Western印迹法检测脑组织 p-Akt、Bax、Bcl-2蛋白的表达。结果 与HIE组相比,R1组、R2组大鼠异常行为减少、体重增长率明显增加(P<0.05)。CaSR阳性细胞数R1组(22.00±1.79)与R2组(20.83±2.04)较HIE组(25.67±1.21)减少(P<0.05);p-Akt阳性细胞数R1组(25.00±1.41)与R2组(27.33±1.86)较HIE组(14.33±2.16)增多(P<0.05);p-Akt光密度值R1组(328.33±24.86)与R2组(339.67±23.77)较HIE组(120.83±17.43)增加(P<0.05)。与HIE 组相比,R1组与R2组Bax表达减少(P<0.05),Bcl-2表达增加(P<0.05)。R1组与R2组相比,各项指标差异无统计学意义(P>0.05)。各组间Akt的表达差异无统计学意义(P>0.05)。结论 远隔缺血后处理对新生大鼠脑组织损伤的保护作用可能与脑组织CaSR表达降低和激活Akt/Pi3k通路有关。
Abstract
Objective To explore the effect of remote ischemic postconditioning (RIPC) on calcium sensing receptorl(CaSR) expression and Akt/Pi3k pathway in brain tissue of neonatal rats with hypoxic-ischemic encephalopathy(HIE), in order to provide theoretical evidence for the treatment of HIE in neonates. Methods Totally 120 newborn Wistar rats aged 7 days were randomly divided into sham operation group (S group), hypoxic-ischemic encephalopathy group (HIE group), RIPC for 5 minutes group (R1 group) and RIPC for 10 minutes group (R2 group).The behaviors of rats in each group were observed and the body weight was measured.The morphological changes were observed by HE staining, and the ultrastructure of cells was observed by electron microscopy.The expressions of CaSR, Bax, Bcl-2,p-Akt and Akt protein were detected by immunohistochemistry, and the expression of p-Akt, Bax and Bcl-2 protein was detected by western blotting. Results Compared with HIE group, rats in R1 group and R2 group showed reduced abnormal behaviors, and the growth rate of body weight increased significantly (P<0.05).The number of CaSR positive cells in R1 group (22.00 ±1.79) and R2 group (20.83 ±2.04) was significantly lower than that in HIE group (25.67±1.21) (P < 0.05).The number of p-Akt positive cells in R1 group (25.00±1.41) and R2 group (27.3 3±1.86) was significantly more than that in HIE group (14.33 ±2.16) (P<0.05).The p-Akt optical density value of R1 group (328.33 ±24.86) and R2 group (339.67± 23.77) was significantly higher than that of HIE group (120.83 ±17.43) (P<0.05).Compared with HIE group, Bax expression level was decreased in R1 group and R2 group (P<0.05), while Bcl-2 expression level was increased (P<0.05).However, there were no significant differences on the indexes between R1 group and R2 group (P>0.05).And the Akt expression level was not significantly different among the four groups (P>0.05). Conclusion RIPC plays a protective role in neonatal rat brain tissue injury, probably because of the decreasing expression of CaSR in brain tissue and the activation of Akt/Pi3k pathway.
关键词
远隔缺血后处理 /
缺氧缺血性脑病 /
钙敏感受体 /
Akt/Pi3k通路
Key words
remote ischemia postconditioning /
hypoxic-ischemic encephalopathy /
calcium sensing receptor /
Akt/pi3k pathway
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基金
国家自然科学基金(81300122); 黑龙江省青年科学基金(QC2016130)
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