吗啡对小鼠室管膜上皮、中央灰质及海马结构中巢蛋白表达的影响

Influence of morphine on expression of nestin in ependymal epithelia,central gray and hippocampal formations of mice

目的:观察吗啡对小鼠脑组织巢蛋白(nestin)表达的影响。方法:20只小鼠随机分为对照组和实验组(n=10)。对照组每只小鼠每日皮下注射生理盐水0.1 ml,实验组每只小鼠每日注射吗啡0.1 ml(1 mg)。30 d后处死小鼠,开颅取脑并制成石蜡切片,免疫组织化学法观察不同部位脑组织nestin的表达,用图像分析系统进行分析。结果:对照组的室管膜上皮、中央灰质、室周灰质和海马结构中巢蛋白呈弱阳性表达,平均灰度值为150.982±13.31;其中有5类巢蛋白阳性细胞:第一类为室管膜上皮的基底细胞,第二类分布于室周灰质和中央灰质的深层,第三类主要分布于中央灰质的浅层、下托、海马旁回和内嗅区皮质的Ⅱ、Ⅲ层,第四类常见于顶盖与下托,第五类位于顶盖、海马、齿状回、海马旁回和内嗅区皮质的第V层;下托和内嗅区的阳性神经细胞数为(7.20±1.23)/mm^2。实验组上述部位脑组织巢蛋白呈阳性表达,平均灰度值为133.03±22.28,上述5类细胞的密度增加,下托和内嗅区的强阳性神经细胞数为(10.50±1.43)/mm^2。两组平均灰度值和阳性神经细胞数均有显著差异(P<0.05)。与对照组相比,实验组小鼠室管膜上皮细胞增生;中央灰质的浅层、海马旁回和内嗅区皮质第Ⅰ、Ⅱ、Ⅲ层内出现了第二类细胞;海马锥体细胞层第三类细胞增加;顶盖和下托内第四类细胞增加。结论:吗啡可能通过促进小鼠室管膜上皮细胞、中央灰质、室周灰质及海马结构巢蛋白的表达促进神经细胞的增殖、分化与迁移。

Objective：To study the influence of morphine on the expression of nestin in the ependymal epithelia,central gray and hippocampal formations in mice. Methods： Twenty health mice were evenly randomized into control group and experiment group. Mice in the control group were injected with normal saline （0. 1 ml daily） and those in the experimental group were injected with morphine （0. 1 ml, 1 mg daily） . Thirty days later, the mice brain samples were harvested and made into paraffin sections. ]mmumohistochemical ABC technique was used to observe the expression of nestin under light microscope. The images were analyzed with the image analytical system. Results： In the control group, the ependymal epithelia,the central gray, the periventricular gray substances and the hippocampal formations had weak expression of the nestin, with a mean gray scale of 150. 98 ±13.31; there were 5 kinds of nestin-positive cells： （1） the basal cells of ependymal epithelium, （2） cells distributed in the periventricular gray substance and the deep lamella of central gray, （3） cells distributed in the superficial lamella of central gray, the subiculum,the parahippocampal gyrus and the cortex in Ⅱ, Ⅲ layers of the entorhinal area, （4） cells frequently seen in the tectum of midbrain and the subiculum,and （5） cells distributed in the rectum of midbrain, the hippocampus,gyrus dentatus, parahippocampal gyrus and the cortex in V layer of the entorhinal area; the density of nestin in the subiculum and entorhinal area was （7.20 ±1.23） mm^2. In the experiment group, the ependymal epithelia, the central gray, the periventricular gray substances and the hippocampal formations had positive expression of the nestin, with the mean gray scale being 133.03±22.28; the density of the above-mentioned 5 kinds of cells increased; the density of nestin in the subiculum and entorhinal area was （10. 50±1.43） mm^2. The mean value of gray scale and nestin-positive neurons were significantly different between the 2 groups （P〈0.05）. The cellular proliferation was seen in the ependymal epithelia. The second kind of cells appeared in the superficial lamella of central gray, parahippocampal gyrus and the cortex in Ⅰ, Ⅱand Ⅲ layers of the entorhinal area. The third kind of cells increased in the hippocampal pyramidal layer and the fourth kind of cells increased in the tectum of midbrain and the subiculum. Conclusion： Morphine can promote nestin expression in the ependymocyte, the central gray,the periventricular gray substance and the hippocampal formation; it can also promote the proliferation,differentiation and migration of the ependymocytes and neurocytes.