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目的探讨单一连续应激(SPS)大鼠内侧前额皮质(mPFC)磷酸化细胞外信号调节激酶(pERK1/2)和c-fos表达的变化。方法将45只雄性Wistar大鼠随机分为对照组、应激组和干预组。应激组和干预组大鼠接受SPS,干预组大鼠接受SPS前30min前额皮质局部注射ERK抑制剂2′-氨基-3′-甲氧黄酮(PD98059)。利用开场实验、高架十字迷宫实验和Morris水迷宫实验检测行为学改变,免疫组织化学和Western blotting检测pERK1/2表达,RT-PCR检测c-fos mRNA表达。结果接受SPS后,大鼠焦虑样行为增强,空间记忆能力减退。应激组大鼠mPFC部位pERK1/2阳性细胞吸光度、pERK1/2蛋白表达量和c-fos mRNA相对表达量分别为51.54±5.41、89.61±3.25和0.91±0.13,显著高于对照组的12.18±1.61、34.22±5.83和0.13±0.03(P<0.01),干预组为26.26±1.42、60.59±5.88和0.35±0.11,显著低于应激组(P<0.01)。结论 PD98059可以阻抑SPS所引起的大鼠行为学改变和mPFC部位pERK1/2及c-fos mRNA表达增高,提示ERK1/2磷酸化在SPS过程中具有重要作用。
Objective To investigate the changes of phosphorylated extracellular signal-regulated kinase (pERK1 / 2) and c-fos expression in medial prefrontal cortex (mPFC) in single continuous stress rats. Methods Forty-five male Wistar rats were randomly divided into control group, stress group and intervention group. Rats in the stress group and the intervention group received SPS. Rats in the intervention group were injected with 2’-amino-3’-methoxy-flavone (PD98059), an ERK inhibitor, into the prefrontal cortex 30 min before receiving SPS. Behavioral changes were detected by open experiment, elevated plus maze test and Morris water maze test. The expression of pERK1 / 2 was detected by immunohistochemistry and Western blotting. The expression of c-fos mRNA was detected by RT-PCR. Results After receiving SPS, anxiety-like behaviors of rats increased, and spatial memory decreased. The expression of pERK1 / 2 protein, the expression of pERK1 / 2 and the relative expression of c-fos mRNA were 51.54 ± 5.41,89.61 ± 3.25 and 0.91 ± 0.13 in mPFC, respectively, which were significantly higher than those in control group (12.18 ± 1.61,34.22 ± 5.83 and 0.13 ± 0.03 (P <0.01). The intervention group was 26.26 ± 1.42,60.59 ± 5.88 and 0.35 ± 0.11, which was significantly lower than that of the stress group (P <0.01). Conclusion PD98059 can inhibit behavioral changes induced by SPS and increase the expression of pERK1 / 2 and c-fos mRNA in mPFC, suggesting that ERK1 / 2 phosphorylation plays an important role in the process of SPS.