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摘除颈上神经节对成年大鼠海马基质金属蛋白酶9和水通道蛋白9 表达的影响
赵倩楠 丁艳平* 邵宝平 张晋平 刘健锋 贾凌云
解剖学报 ›› 2015, Vol. 46 ›› Issue (2) : 158-163.
摘除颈上神经节对成年大鼠海马基质金属蛋白酶9和水通道蛋白9 表达的影响
The influence of superior cervical ganglionectomy on expression of matrix metalloproteinase-9 and aquaporins-9 in the hippocampus in adult rats
目的 观察摘除颈上神经节（SCG）对成年大鼠海马基质金属蛋白酶9（MMP9）和水通道蛋白9（AQP9）表达的影响，探索交感神经对脑组织的调节作用及机制。 方法 40只成年健康雄性Wistar大鼠分为对照组和手术组，手术组大鼠摘除双侧SCG；用免疫组织化学法对海马区MMP9及AQP9蛋白的表达进行定位，Western blotting技术对海马区MMP9及AQP9蛋白的表达进行定量分析，RTPCR技术测定海马区MMP9 mRNA及AQP9 mRNA的表达变化。 结果 MMP9免疫阳性反应产物表达于神经元和神经胶质细胞，手术组海马区 MMP9的表达明显强于对照组，CA1区和CA3 区的阳性神经元在锥体细胞层散在分布，齿状回（DG）区的强阳性细胞主要位于颗粒细胞层；手术组海马MMP9蛋白表达和MMP9 mRNA表达水平明显升高，和对照组相比差异极显著（P<0.01 )。AQP9免疫阳性反应产物在神经元、神经胶质细胞和血管均有表达，手术组 AQP9的表达明显强于对照组，DG区的强阳性细胞主要位于颗粒细胞下层的新生细胞，且室管膜细胞中也有AQP9的高表达；手术组AQP9蛋白表达和AQP9 mRNA表达水平均明显增强，与对照组相比差异极显著（P<0.01）。 结论 摘除成年大鼠SCG 15d 后，海马MMP9、AQP9的表达明显上调，表明脑部去交感所引起的脑供血的改变会对海马多个因子的表达产生影响而发挥调节作用。
Objective To study the expression of matrix metalloproteinase-9 (MMP9), aquaporins-9 (AQP9）in the hippocampus after the superior cervical ganglionectomy in adult rats, and to explore the regulate mechanism of sympathetic ganglion on the brain tissue. Methods A total of 40 healthy adult male Wistar rats were divided into control and operation groups. The surgery of superior cervical ganglionectomy was perfomed in the operation group. Immunohistochemistry (IHC) was used to localize the expression of MMP9 and AQP9 in the hippocampus. Western-blotting was used to quantify the expression change of MMP9 and AQP9. RT-PCR was used to detect the variation of MMP9 mRNA and AQP9 mRNA in the hippocampus. Results MMP9 immunohistochemical product was mainly expressed in neurons and glial cells. The expression of MMP9 in hippocampus of the operation group was significantly stronger than the control group. The few positive neurons in CA1 and CA3 were scattered in the pyramidal cell layer , and strongly positive cells were mainly located in the granular layer of dentat gyrus(DG) region. Compared to the control group, the MMP9 protein expression and MMP9 mRNA expression levels in the hippocampus of the operate group were significantly increased (P<0.01). AQP9 immunohistochemical product was mainly expressed in neurons, glial cells, and blood vessels; the expression of AQP9 in the hippocampus of the operation group was significantly stronger than the control group. The positive cells were mainly located in new born cells of subgranular zone of DG region and ependymal cells. Compared to the control group, the AQP9 protein expression and AQP9 mRNA expression levels in the hippocampus of the operate group were significantly increased (P<0.01) than control group. Conclusion The expressions of MMP9 and AQP9 are significantly increased in the hippocampus after 15 days of the superior cervical ganglionectomy surgery, indicating that removal of sympathetic ganglions may change cerebral blood supply and further affect the expression of some related factors in the hippocampus.
颈上神经节 / 海马 / 基质金属蛋白酶9 / 水通道蛋白9 / 免疫印迹法 / 大鼠
Superior cervical ganglion / Hippocampus / Matrix metalloproteinase-9 / Aquaporin-9 / Western blotting / Rat
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