标题：改良大鼠脊髓缺血再灌注损伤模型的再评价及应用研究

      作者：张志琴 1，王振飞 2，何大伟 1，陆轲 3，郝彦明 3，李翀 3    1.昆山市第一人民医院生物样本库，江苏 昆山 215316；2.徐州市中心医院骨科，江苏 徐州 221009；3.昆山市第一人民医院骨科，江苏 昆山 215316

      卷次： 2022年33卷6期

      【摘要】 目的 建立并评价一种改良大鼠脊髓缺血再灌注损伤(SCII)模型。方法 选取清洁级雄性SD大鼠36只(300~400 g)，经腹腔注射麻醉药物，麻醉满意后打开腹腔并夹闭右肾动脉上极腹主动脉近心端1 h，分别再灌注不同时间制作改良大鼠SCII模型。将实验动物随机分为A1组(血管造影剂对照组，给予正常大鼠左心室灌注造影剂Microfil)、A2组(实验对照组，夹闭腹主动脉后左心室灌注Microfil)、B1组(假手术组)、B2组(缺血60 min再灌注24 h)、B3组(缺血60 min再灌注48 h)、B4组(缺血90 min再灌注24 h)和B5组(缺血90 min再灌注48 h)。A组大鼠同步辐射CT三维重建夹闭腹主动脉前后脊髓动脉血供分布情况；B组采用BBB分级法评价大鼠下肢运动功能变化；HE染色和尼氏染色观察损伤脊髓组织随缺血时间延长病理动态变化。结果 同步辐射CT三维重建结果显示：夹闭大鼠右肾动脉上腹主动脉近心端可以阻断T13以下节段脊髓大部分动脉血供，但不能完全阻断；与对照组比较，BBB评分从B2至B5逐渐降低，各组间比较差异具有统计学意义(P<0.05)；尼氏染色及HE染色显示B1组无明显改变，再灌注时间同样的情况下，缺血时间的长短与脊髓病理学改变程度以及脊髓灰质前角神经元改变成正比，表现为神经元尼氏体淡染或消失。结论 经过改良的SCII模型可以模拟不同程度的缺血再灌注损伤，缺血范围较传统方法更广，经济可靠，病理改变稳定。
      【关键词】 脊髓缺血再灌注损伤；大鼠模型；血管造影；动脉分布；病理
      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2022）06—0681—04

Re-evaluation and application research of an improved spinal cord ischemia-reperfusion injury model in rats.ZHANG Zhi-qin 1, WANG Zhen-fei 2, HE Da-wei 1, LU Ke 3, HAO Yan-ming 3, LI Chong 3.
1. Biobank, the First People'sHospital of Kunshan, Kunshan 215316, Jiangsu, CHINA; 2. Department of Orthopedics, Xuzhou Central Hospital, Xuzhou221009, Jiangsu, CHINA; 3. Department of Orthopedics, the First People's Hospital of Kunshan, Kunshan 215316, Jiangsu,CHINA
【Abstract】 Objective To establish and evaluate an improved spinal cord ischemia-reperfusion injury (SCII)model in rats. Methods A total of 36 male SD rats (300~400 g) were selected. After intravenous anesthesia, a model ofacute SCII was established: the aorta was clipped above the right renal artery near the heart using a 50-g aneurysm clip,and each animal underwent spinal cord ischemia for 1 hour. The experimental animals were randomly divided into 7groups: A1 group (Microfil control group): normal rats were perfused with contrast agent Microfil into left ventricle; A2group (Microfil experimental group): normal rats were perfused with Microfil into left ventricle after aortic was clipped;B1 group (sham group); B2, B3 group (spinal cord ischemia for 60 min and reperfusion for 24 h or 48 h, respectively);B4, B5 group (spinal cord ischemia for 90 min and reperfusion for 24 h or 48 h, respectively). The distribution of spinalcord blood supply in A group were reconstructed by synchrotron radiation CT. The locomotor recovery after SCII wasscored by Basso-Beattie-Bbresnahan (BBB) open-field locomotor rating scale. The pathological changes in spinal cordwere dynamically observed by HE staining and Nissl staining. Results The results of synchrotron radiation CT recon-struction showed that the spinal blood supply can be blocked below T13, but can’t be blocked completely. Compared withthe control group, BBB score decreased gradually from B2 group to B5 group, and the difference between the groupswere statistically significant (P<0.05). HE staining and Nissl staining showed that there was no significant change ingroup B1. Under the same reperfusion time, the length of ischemia time was directly proportional to the degree of patho-logical changes of spinal cord and the changes of neurons in the anterior horn of spinal gray matter, which showed thatNissl bodies of neurons were lightly stained or disappeared. Conclusion The improved SCII model can simulate differ-ent degrees of SCII, and the range of ischemia is wider than the traditional method. The model is economical and reliableand the pathological change is stable.
      【Key words】 Spinal cord ischemia-reperfusion injury; Rat model; Angiography; Arterial distribution; Pathology