脂多糖干预对大鼠周围神经损伤后瓦勒变性的影响
发布时间：2019-08-01 18:58
【摘要】：目的探讨脂多糖对于大鼠坐骨神经损伤瓦勒变性早期髓鞘碎片清除的影响。方法将50只Wistar大鼠随机分成假手术组(10只),模型组(20只)和脂多糖LPS组(20只),LPS组及模型组横断大鼠右侧坐骨神经后,行神经外膜端端吻合;假手术组仅游离出坐骨神经,然后关闭切口。LPS组大鼠在神经断端显微注射LPS(2 g/L)1μL,模型组及假手术组大鼠注射同等体积生理盐水。于术后1.5、24 h和7 d取术侧坐骨神经。实时定量PCR(qRT-PCR)检测坐骨神经中白介素1β(IL-1β)mRNA、单核细胞趋化蛋白-1(MCP-1)mRNA水平;免疫荧光法检测坐骨神经中CD68+巨噬细胞的表达;HE染色观察坐骨神经的病理变化;油红O染色观察坐骨神经脱髓鞘程度;LFB染色观察坐骨神经髓鞘变化;坐骨神经功能指数(SFI)评价大鼠运动功能的恢复情况。结果实时定量PCR显示,与假手术组相比,术后1.5 h模型组IL-1βmRNA和MCP-1 mRNA的表达均明显升高(P0.001,P0.001),与模型组相比,术后1.5 h LPS组IL-1βmRNA和MCP-1mRNA的表达明显升高(P0.001,P0.001)。术后24 h模型组IL-1βmRNA和MCP-1m RNA的表达均明显升高(P0.001,P0.001),与模型组相比,术后24h LPS组IL-1βmRNA和MCP-1 mRNA的表达明显升高(P0.01,P0.01)。免疫荧光可见,与模型组相比,术后7 d LPS组中CD68+细胞表达显著上调(P0.05)。术后7 d坐骨神经HE染色可见,LPS组坐骨神经断端较多炎性细胞浸润,许旺细胞增殖活跃,模型组神经断端炎性细胞和许旺细胞较少。术后7 d坐骨神经ORO染色可见,与模型组相比,LPS组断端远侧脱髓鞘程度较高。术后7 d坐骨神经LFB染色可见,模型组和LPS组坐骨神经断端均出现脱髓鞘反应,但与模型组相比,LPS组神经断端残余髓鞘碎片明显减少(P0.05)。SFI显示,与模型组相比,LPS组大鼠在术后10、20、30、40和50 d分别不同程度升高,术后20 d明显增高,差异有显著性(P0.05)。结论脂多糖通过激活固有免疫系统加快大鼠坐骨周围神经损伤后瓦勒变性早期髓鞘碎片的清除。
[Abstract]:Objective to investigate the effect of lipopolysaccharide (lipopolysaccharide) on the removal of myelin sheath fragments in the early stage of Vall degeneration after sciatic nerve injury in rats. Methods 50 Wistar rats were randomly divided into three groups: model group (n = 20), lipopolysaccharide LPS group (n = 20) and lipopolysaccharide LPS group (n = 20). The rats in the model group were treated with end-to-end anastomosis after transection of the right sciatica. The rats in the pseudo-operation group were only free of the sciatica and then closed the incision. the rats in the LPs group were microinjected with LPS (2 g 路L) 1 渭 L, and the rats in the model group and the pseudo-operation group were injected with the same volume of normal saline. The sciatica nerve was taken 24 hours and 7 days after operation. Real-time quantitative PCR (qRT-PCR) was used to detect the level of IL-1 尾 (IL-1 尾) mRNA, monocyte chemotactic protein-1 (MCP-1) mRNA in the sciatica, the expression of CD68 macrophages in the sciatica was detected by immunofluorescence, the pathological changes of the sciatica were observed by HE staining, the demyelination degree of the sciatica was observed by oil red O staining, and the myelin sheath of the sciatica was observed by LFB staining. The recovery of motor function in rats was evaluated by (SFI). Results Real-time quantitative PCR showed that the expression of IL- 1 尾 mRNA and MCP-1mRNA in the model group was significantly higher than that in the pseudo operation group at 1.5 h after operation (P0.001P0.001), and the expression of IL- 1 尾 mRNA and MCP-1mRNA in the LPS group was significantly higher than that in the model group at 1.5h after operation (P0.001P0.001). The expression of IL- 1 尾 mRNA and MCP-1m RNA in model group was significantly higher than that in model group 24 hours after operation (P0.001P0.001). Compared with model group, the expression of IL- 1 尾 mRNA and MCP-1 mRNA in LPS group was significantly higher than that in model group 24 hours after operation (P0.01, P0.01). Compared with the model group, the expression of CD68 cells in LPS group was significantly up-regulated on the 7th day after operation (P 0.05). On the 7th day after operation, HE staining showed that there were more inflammatory cells infiltrated and Schwann cells proliferated at the severed end of the sciatica in the LPS group, but fewer inflammatory cells and Schwann cells were found in the model group. On the 7th day after operation, ORO staining showed that the degree of demyelination at the distal end of the severed nerve in the LPS group was higher than that in the model group. On the 7th day after operation, LFB staining showed demyelination in both the model group and the LPS group, but compared with the model group, the residual myelin sheath fragments at the broken end of the nerve in the LPS group were significantly lower than those in the model group (P 0.05). LFB showed that compared with the model group, the LPS group increased at 10, 20, 40 and 50 days after operation, and increased significantly at the 20th day after operation (P 0.05). Conclusion lipopolysaccharide accelerates the clearance of myelin sheath fragments in the early stage of Vall degeneration after ischial nerve injury in rats by activating the innate immune system.
【作者单位】： 青岛大学基础医学院;青岛阜外心血管病医院;
【基金】：山东省高等学校科技计划(编号J14LK10,J16LK04)
【分类号】：R745
本文编号：2521937
[Abstract]:Objective to investigate the effect of lipopolysaccharide (lipopolysaccharide) on the removal of myelin sheath fragments in the early stage of Vall degeneration after sciatic nerve injury in rats. Methods 50 Wistar rats were randomly divided into three groups: model group (n = 20), lipopolysaccharide LPS group (n = 20) and lipopolysaccharide LPS group (n = 20). The rats in the model group were treated with end-to-end anastomosis after transection of the right sciatica. The rats in the pseudo-operation group were only free of the sciatica and then closed the incision. the rats in the LPs group were microinjected with LPS (2 g 路L) 1 渭 L, and the rats in the model group and the pseudo-operation group were injected with the same volume of normal saline. The sciatica nerve was taken 24 hours and 7 days after operation. Real-time quantitative PCR (qRT-PCR) was used to detect the level of IL-1 尾 (IL-1 尾) mRNA, monocyte chemotactic protein-1 (MCP-1) mRNA in the sciatica, the expression of CD68 macrophages in the sciatica was detected by immunofluorescence, the pathological changes of the sciatica were observed by HE staining, the demyelination degree of the sciatica was observed by oil red O staining, and the myelin sheath of the sciatica was observed by LFB staining. The recovery of motor function in rats was evaluated by (SFI). Results Real-time quantitative PCR showed that the expression of IL- 1 尾 mRNA and MCP-1mRNA in the model group was significantly higher than that in the pseudo operation group at 1.5 h after operation (P0.001P0.001), and the expression of IL- 1 尾 mRNA and MCP-1mRNA in the LPS group was significantly higher than that in the model group at 1.5h after operation (P0.001P0.001). The expression of IL- 1 尾 mRNA and MCP-1m RNA in model group was significantly higher than that in model group 24 hours after operation (P0.001P0.001). Compared with model group, the expression of IL- 1 尾 mRNA and MCP-1 mRNA in LPS group was significantly higher than that in model group 24 hours after operation (P0.01, P0.01). Compared with the model group, the expression of CD68 cells in LPS group was significantly up-regulated on the 7th day after operation (P 0.05). On the 7th day after operation, HE staining showed that there were more inflammatory cells infiltrated and Schwann cells proliferated at the severed end of the sciatica in the LPS group, but fewer inflammatory cells and Schwann cells were found in the model group. On the 7th day after operation, ORO staining showed that the degree of demyelination at the distal end of the severed nerve in the LPS group was higher than that in the model group. On the 7th day after operation, LFB staining showed demyelination in both the model group and the LPS group, but compared with the model group, the residual myelin sheath fragments at the broken end of the nerve in the LPS group were significantly lower than those in the model group (P 0.05). LFB showed that compared with the model group, the LPS group increased at 10, 20, 40 and 50 days after operation, and increased significantly at the 20th day after operation (P 0.05). Conclusion lipopolysaccharide accelerates the clearance of myelin sheath fragments in the early stage of Vall degeneration after ischial nerve injury in rats by activating the innate immune system.
【作者单位】： 青岛大学基础医学院;青岛阜外心血管病医院;
【基金】：山东省高等学校科技计划(编号J14LK10,J16LK04)
【分类号】：R745
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