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目的:本研究利用新生大鼠缺氧缺血性脑损伤(hypoxic—ischemic brain damage,H1BD)动物模型,观察应用肌苷(Inosine)前后脑组织哺乳动物Ste20样蛋白激酶-3b(mammalian Ste20-like protein kinase-3b,Mst3b)水平的改变,探讨新生大鼠HIBD早期应用肌苷后Mst3b水平的改变与脑损伤的关系。方法:7日龄Wistar大鼠随机分为假手术组(对照组)、模型组和用药组(肌苷组),分别给予腹腔注射同等剂量的生理盐水(对照组、模型组)和0.5%肌苷(肌苷组),给药后48h采集标本。嗜银和HE染色法光镜下观察神经纤维和脑神经细胞的病理变化:免疫组化和Western Blot方法检测脑组织Mst3b水平的表达。结果:(1)对照组HE染色后脑组织无坏死,神经细胞形态无异常,轮廓清晰可见,核仁核膜明显,大小无变化,未见明显损伤性改变:模型组HE病理检测毛细血管出血明显可见,核固缩、核碎裂明显,提示模型建立成功。(2)对照组脑神经细胞染色分布均匀,柄突、轴突走行分布清晰:模型组嗜银染色示神经纤维明显增粗,部分神经纤维可见缠结现象;肌苷组神经纤维略显增粗,未见明显的神经纤维缠结。(3)免疫组化和Western Blot检测Mst3b值:肌苷组高于模型组(P<0.01),模型组高于对照组(p<0.05)。结论:HIBD新生大鼠早期应用肌苷后Mst3b的表达增加,肌苷对脑神经细胞具有保护作用。
Objective: In this study, animal models of hypoxic-ischemic brain damage (HIBD) in neonatal rats were used to observe the effects of Inosine on mammalian Ste20-like mammals such as Ste20-like protein kinase-3b, Mst3b) levels in neonatal rats to explore the early application of inosine HIBD Mst3b levels and brain damage. Methods: The 7-day-old Wistar rats were randomly divided into sham-operation group (control group), model group and administration group (inosine group). The rats were given intraperitoneal injection of normal saline (control group, model group) Glycosides (inosine group), specimens collected 48h after administration. The pathological changes of nerve fibers and neurons were observed by silver staining and HE staining. The expression of Mst3b in brain tissue was detected by immunohistochemistry and Western Blot. Results: (1) There was no necrosis in the brain tissue after HE staining in the control group, the morphology of the nerve cells was normal, and the outline was clear. The nuclear membrane of the nucleus was obvious and its size did not change. No obvious damage was found. Visible, nuclear pyknosis, nuclear fragmentation obvious, suggesting that the model was established. (2) In the control group, the distribution of neuronal cells in the control group was uniform and the processes of axons and axons were well distributed. The argyrophilic staining in the model group showed that the nerve fibers were obviously thicker and some of the nerve fibers showed tangles; , No obvious nerve fiber tangle. (3) Mst3b was detected by immunohistochemistry and Western Blot: the inosine group was higher than the model group (P <0.01), the model group was higher than the control group (p <0.05). Conclusion: The expression of Mst3b is increased in neonatal HIBD neonatal rats after inosine administration. Inosine has a protective effect on neurons.