多聚肌苷酸多聚胞苷酸对大鼠脊髓缺血再灌注损伤后炎性反应的脊髓保护作用

发布时间：2019-04-10 09:38

【摘要】：目的多聚肌苷酸多聚胞苷酸(Poly-IC)在中枢神经系统损伤与修复中起重要作用。文中探讨Toll样受体3(TLR3)配体Poly-IC对大鼠脊髓缺血再灌注损伤(SCIRI)后炎性反应的影响。方法 72只成年健康雄性SD大鼠随机数字表法分为假手术组、缺血再灌注组、Poly-IC组,每组24只。假手术组只做腹腔打开,缺血再灌注组、Poly-IC组打开腹腔后,夹闭左肾动脉起始水平近心端及左、右髂总动脉分叉水平腹主动脉,60min后再灌注,关闭腹腔,分别腹腔给予0.1mL等渗盐水、1.25μg/g Poly-IC。3组大鼠分别于造模后6、24、48 h检测BBB神经功能评分,酶联免疫分析法检测各时间点脊髓组织中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及干扰素β(IFN-β)的含量;各组造模后48 h行免疫组化检测核转录因子(NF-κB)及IL-10的表达,TTC染色显示脊髓组织缺血坏死面积,HE染色光镜下观察脊髓组织形态学改变。结果各组造模48h时,与假手术组比较,缺血再灌注组、Poly-IC组BBB评分均降低(P0.01);与缺血再灌注组相比,Poly-IC组BBB评分显著增加[(3.80±0.75)分vs(9.40±0.49)分,P0.01],缺血再灌注组脊髓灰质出现广泛的变性神经元,可见散在的出血和凝血灶,Poly-IC组脊髓组织神经元损伤坏死数量减少,细胞核形态基本正常,仍可见少量细胞肿胀。TTC染色结果显示经过Poly-IC治疗后坏死面积明显比缺血再灌注组要小。NF-κB免疫组化检测脊髓组织NF-κB的活性,缺血再灌注组可见大量神经元细胞表达,Poly-IC组阳性表达细胞数量明显减少。检测脊髓组织IL-10的表达,缺血再灌注组可见少量神经元细胞细胞质表达细胞,Poly-IC组阳性表达细胞数量明显增加。与假手术组相比,缺血再灌注组和Poly-IC组TNF-α、IL-1β、IFN-β均明显升高(P0.05);与缺血再灌注组相比,Poly-IC组TNF-α、IL-1β含量降低(P0.05),IFN-β含量明显增高(P0.05)。结论 PolyIC能减轻后炎性反应,对脊髓有一定的保护作用。
[Abstract]:Objective Polyinosine polycytidylate (Poly-IC) plays an important role in the injury and repair of central nervous system (CNS). To investigate the effect of Toll-like receptor 3 (TLR3) ligand Poly-IC on inflammatory response after spinal cord ischemia-reperfusion injury in rats with (SCIRI). Methods 72 adult male SD rats were randomly divided into sham-operated group, ischemia-reperfusion group and Poly-IC group with 24 rats in each group. After opening abdominal cavity in sham operation group, ischemia-reperfusion group and Poly-IC group, the initial level of left renal artery was closed near the heart end and left, the right common iliac artery bifurcation level abdominal aorta, 60min and reperfusion, closed the abdominal cavity, and closed the abdominal cavity at the beginning level of left renal artery and left, right common iliac artery bifurcation level, and closed abdominal cavity after 60min. The rats in the 1.25ug / g Poly-IC.3 group were treated with 0.1mL isoosmotic saline respectively. The BBB nerve function score was measured at 6,24,48 hours after the establishment of the model, and the tumor necrosis factor 伪 (TNF- 伪) in the spinal cord tissue was detected by enzyme linked immunosorbent assay (Elisa) at each time point. The contents of interleukin-1 尾 (IL-1-尾) and interferon-尾 (IFN- 尾); The expression of NF- 魏 B and IL-10 was detected by immunohistochemistry at 48 h after modeling in each group. The ischemic necrosis area of spinal cord was shown by TTC staining, and the morphological changes of spinal cord tissue were observed by HE staining under light microscope. Results compared with sham-operated group, BBB scores in ischemia-reperfusion group and Poly-IC group were lower than those in sham-operation group at 48h after modeling (P0.01). Compared with the ischemia-reperfusion group, the BBB score of the Poly-IC group increased significantly [(3.80 卤0.75) vs (9.40 卤0.49), P0.01]. Extensive degeneration neurons appeared in the gray matter of the spinal cord in the ischemia-reperfusion group. In the scattered hemorrhage and coagulation foci, the number of injured and necrotic neurons in the spinal cord tissue of the Poly-IC group decreased, and the shape of the nucleus was basically normal. The results of TTC staining showed that the necrotic area after Poly-IC treatment was significantly smaller than that of ischemia-reperfusion group. The activity of NF- 魏 B in spinal cord tissue was detected by immunohistochemical staining of NF-魏 B. A large number of neurons were expressed in ischemia-reperfusion group, but the number of positive cells in Poly-IC group was significantly decreased. By detecting the expression of IL-10 in spinal cord, a small number of cytoplasm-expressing cells were found in ischemia-reperfusion group, and the number of positive cells in Poly-IC group was significantly increased. Compared with sham operation group, TNF- 伪, IL-1 尾 and IFN- 尾 in ischemia reperfusion group and Poly-IC group were significantly higher than those in sham operation group (P0.05). Compared with ischemia-reperfusion group, the content of TNF- 伪 and IL-1 尾 in Poly-IC group decreased (P0.05), and the content of IFN- 尾 increased significantly (P0.05). Conclusion PolyIC can alleviate the post-inflammatory reaction and protect the spinal cord to a certain extent.
【作者单位】：新乡医学院第一附属医院骨科;新乡医学院生物医学工程学院;
【基金】：河南省神经修复重点实验室开放课题资助项目(HNSJXF-2016-011)
【分类号】：R651.2

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