脂多糖干預對大鼠周圍神經損傷后瓦勒變性的影響
發(fā)布時間：2019-08-01 18:58
【摘要】：目的探討脂多糖對于大鼠坐骨神經損傷瓦勒變性早期髓鞘碎片清除的影響。方法將50只Wistar大鼠隨機分成假手術組(10只),模型組(20只)和脂多糖LPS組(20只),LPS組及模型組橫斷大鼠右側坐骨神經后,行神經外膜端端吻合;假手術組僅游離出坐骨神經,然后關閉切口。LPS組大鼠在神經斷端顯微注射LPS(2 g/L)1μL,模型組及假手術組大鼠注射同等體積生理鹽水。于術后1.5、24 h和7 d取術側坐骨神經。實時定量PCR(qRT-PCR)檢測坐骨神經中白介素1β(IL-1β)mRNA、單核細胞趨化蛋白-1(MCP-1)mRNA水平;免疫熒光法檢測坐骨神經中CD68+巨噬細胞的表達;HE染色觀察坐骨神經的病理變化;油紅O染色觀察坐骨神經脫髓鞘程度;LFB染色觀察坐骨神經髓鞘變化;坐骨神經功能指數(shù)(SFI)評價大鼠運動功能的恢復情況。結果實時定量PCR顯示,與假手術組相比,術后1.5 h模型組IL-1βmRNA和MCP-1 mRNA的表達均明顯升高(P0.001,P0.001),與模型組相比,術后1.5 h LPS組IL-1βmRNA和MCP-1mRNA的表達明顯升高(P0.001,P0.001)。術后24 h模型組IL-1βmRNA和MCP-1m RNA的表達均明顯升高(P0.001,P0.001),與模型組相比,術后24h LPS組IL-1βmRNA和MCP-1 mRNA的表達明顯升高(P0.01,P0.01)。免疫熒光可見,與模型組相比,術后7 d LPS組中CD68+細胞表達顯著上調(P0.05)。術后7 d坐骨神經HE染色可見,LPS組坐骨神經斷端較多炎性細胞浸潤,許旺細胞增殖活躍,模型組神經斷端炎性細胞和許旺細胞較少。術后7 d坐骨神經ORO染色可見,與模型組相比,LPS組斷端遠側脫髓鞘程度較高。術后7 d坐骨神經LFB染色可見,模型組和LPS組坐骨神經斷端均出現(xiàn)脫髓鞘反應,但與模型組相比,LPS組神經斷端殘余髓鞘碎片明顯減少(P0.05)。SFI顯示,與模型組相比,LPS組大鼠在術后10、20、30、40和50 d分別不同程度升高,術后20 d明顯增高,差異有顯著性(P0.05)。結論脂多糖通過激活固有免疫系統(tǒng)加快大鼠坐骨周圍神經損傷后瓦勒變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.
【作者單位】： 青島大學基礎醫(yī)學院;青島阜外心血管病醫(yī)院;
【基金】：山東省高等學�？萍加媱�(編號J14LK10,J16LK04)
【分類號】：R745
本文編號：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.
【作者單位】： 青島大學基礎醫(yī)學院;青島阜外心血管病醫(yī)院;
【基金】：山東省高等學�？萍加媱�(編號J14LK10,J16LK04)
【分類號】：R745
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本文編號：2521937
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