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脑出血(intracerebral hemorrhage, ICH)是临床常见病、多发病,具有发病率、病死率、致残率均高等特点,已成为威胁人类健康的主要疾病之一[1-3]。因此,进行ICH的早诊断、早治疗以及急慢性期病理生理改变的实验研究是非常重要的[4]。目前,大鼠是构建ICH动物模型最常用的动物之一,研究者常通过神经行为学评分,如神经损伤严重缺损评分、平衡木测试、水迷宫测试等方法判断大鼠的神经功能损伤程度,以间接评价大鼠ICH模型构建成功与否,以及评价后期相关干预实验。传统的神经功能损伤评分等方法因其具有间接性和不可避免的主观性,对评估大鼠ICH后神经功能损伤程度尚有一定局限性[5]。Micro-PET/CT能清晰、灵敏地显示脑血肿及其周围神经细胞的代谢情况,明确病变位置、范围和大小,为研究ICH的发病机制、病理转归、生理学改变甚至药物干预提供了有效手段。近年来,国内外已有一些学者利用micro-PET/CT研究ICH[6-7],但有关micro-PET/CT在验证ICH模型中的价值的研究少有报道。本研究应用18F-FDG micro-PET/CT动态观察大鼠ICH模型在不同出血时间点脑血肿体积的变化,直接评价大鼠ICH模型构建成功与否以及后期的恢复情况。
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假手术组大鼠的神经功能损伤评分均为0分。ICH模型组大鼠在ICH模型构建完成、麻醉苏醒后均出现了明显的神经精神症状,表现为精神萎靡不振、食欲欠佳、运动缓慢、左侧肢体瘫痪、行走时向左侧内旋转甚至伴有全身震颤等。ICH后6、24、48 h和3、5、7、14 d,ICH模型组大鼠神经功能损伤评分分别为(2.21±0.30)、(3.51±0.66)、(2.83±0.20)分和(2.12±0.50)、(1.44±0.37)、(1.02±0.25)、(0.51±0.12)分,其中24 h的评分最高,之后随着时间的推移大鼠神经功能开始逐渐恢复。
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(1)视觉分析结果:18F-FDG micro-PET/CT显像示假手术组大鼠大脑的 18F-FDG分布均匀,典型图像见图1。ICH模型组大鼠各个时间点大脑右侧基底节区18F-FDG摄取均减低或缺损(即血肿部位)(图2A),且与大鼠解剖后脑组织血肿区域相对应(图2B)。(2)半定量分析结果:ICH模型组大鼠在ICH后各个时间点micro-PET/CT勾画ROI法测得的脑血肿体积的结果见表1,其中24 h脑血肿体积最大,之后随着出血时间的推移脑血肿体积开始逐渐缩小;24 h脑血肿体积与6、48 h脑血肿体积比较,差异无统计学意义(F=2.27,P>0.05),与3、5、7、14 d比较,差异有统计学意义(F=29.65,P<0.05)。
图 1 假手术组大鼠的18F-FDG micro-PET/CT横断位显像图 显示大鼠大脑18F-FDG分布均匀。假手术组为以0.9%生理盐水代替胶原酶Ⅳ构建的假脑出血模型。FDG为氟脱氧葡萄糖;PET/CT为正电子发射断层显像计算机体层摄影术
Figure 1. 18F-FDG micro-PET/CT transverse imaging of rats in the sham operation group
图 2 脑出血模型组大鼠18F-FDG micro-PET/CT横断位显像图(A)与脑血肿解剖图(B)
Figure 2. 18F-FDG micro-PET/CT transverse imaging (A) and cerebral hematoma anatomical topography (B) of rats in the intracerebral hemorrhage model group
方法 6 h 24 h 48 h 3 d 5 d 7 d 14 d micro-PET/CT
勾画ROI法24.05±3.00 27.19±1.25 25.58±1.57 21.94±0.98 19.88±1.53 18.35±2.11 16.29±1.53 多田公式 23.17±1.93 26.09±1.35 24.64±1.95 21.31±1.32 19.07±1.64 17.29±1.38 15.63±1.98 t 值 1.58 1.18 3.06 1.80 2.21 2.84 1.91 P值 0.21 0.32 0.06 0.17 0.11 0.07 0.15 注:PET/CT为正电子发射断层显像计算机体层摄影术;多田公式为A×B×C/2(A和B分别为血肿最大层面测量的长径和短径;C为血肿切片数) 表 1 2种方法测得的脑出血模型组大鼠在脑出血后不同时间点脑血肿体积的比较[(
) mm3,n=4]$\bar{x}\pm s$ Table 1. Comparison of cerebral hematoma volume measured by two methods in rats in the intracerebral hemorrhage model group at different time points after intracerebral hemorrhage [(
)mm3,n=4]$\bar{x}\pm s $ -
ICH模型组在各个时间点解剖后,大脑右侧基底节区均可见不规则的血肿形成,与18F-FDG micro-PET/CT显示的放射性稀疏及缺损区相对应。而且随着出血后时间的延长,脑血肿体积逐渐缩小。典型的大鼠脑血肿解剖图见图2B。图3示脑组织血肿内大量弥漫分布的红细胞。
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Micro-PET/CT勾画ROI法和多田公式测得的ICH大鼠各个时间点的脑血肿体积的差异均无统计学意义(均P>0.05,表1);2种方法测得的脑血肿体积呈显著正相关(r=0.99,P<0.001)。
18F-FDG micro-PET/CT 在活体大鼠脑出血模型中的诊断价值
Diagnostic value of 18F-FDG micro-PET/CT in rat models of intracerebral hemorrhage in vivo
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摘要:
目的 评价18F-氟脱氧葡萄糖(FDG) micro-PET/CT 对活体大鼠脑出血(ICH)模型的诊断价值。 方法 采用简单随机抽样法选取32只健康雄性成年SD大鼠,其中28只在大鼠大脑右侧基底节区注射0.125 U/μL胶原酶Ⅳ构建大鼠ICH模型(ICH模型组),其余4只以0.9%生理盐水代替胶原酶Ⅳ构建假模型(假手术组)。采用简单随机抽样法将ICH模型组大鼠按脑出血后的时间分为7个组:6、24、48 h和3、5、7、14 d,每组4只。假手术组和7个ICH模型组均行神经功能损伤评分和18F-FDG micro-PET/CT显像,18F-FDG micro-PET/CT勾画感兴趣区(ROI)法和多田公式分别计算ICH模型组各时间点的脑血肿体积。大鼠显像后取脑,观察脑内血肿情况。同一时间点2种方法得出的脑血肿体积的比较采用配对t检验,并将2种方法得出的脑血肿体积行Pearson相关性分析。 结果 假手术组大鼠神经功能损伤评分均为0分;18F-FDG micro-PET/CT显像结果显示大鼠大脑18F-FDG分布均匀。ICH模型组大鼠在出血后6、24、48 h和3、5、7、14 d的神经功能损伤评分分别为(2.21±0.30)、(3.51±0.66)、(2.83±0.20)、(2.12±0.50)、(1.44±0.37)、(1.02±0.25)、(0.51±0.12)分;18F-FDG micro-PET/CT显像结果显示,在各个时间点大鼠大脑右侧基底节区均见18F-FDG摄取减低或缺损;18F-FDG micro-PET/CT勾画ROI法测得的脑血肿体积分别为(24.05±3.00)、(27.19±1.25)、(25.58±1.57)、(21.94±0.98)、(19.88±1.53)、(18.35±2.11)、(16.29±1.53) mm3;多田公式计算的脑血肿体积分别为(23.17±1.93)、(26.09±1.35)、(24.64±1.95)、(21.31±1.32)、(19.07±1.64)、(17.29±1.38)、(15.63±1.98) mm3。2种方法计算所得的ICH模型组大鼠在各个时间点的脑血肿体积间的差异均无统计学意义(t=1.18~3.06,均P>0.05)。2种方法所得的脑血肿体积呈显著正相关(r=0.99,P<0.001)。ICH模型组大鼠解剖后,大脑右侧基底节区均可见不规则血肿形成,与18F-FDG micro-PET/CT显示的放射性稀疏及缺损区相对应。 结论 18F-FDG micro-PET/CT能准确显示ICH后血肿的位置、形态及大小,其可以作为活体验证大鼠ICH模型构建成功与否的新型方法。 -
关键词:
- 脑出血 /
- 正电子发射断层显像术 /
- 体层摄影术,X线计算机 /
- 氟脱氧葡萄糖F18 /
- 大鼠 /
- 模型,动物
Abstract:Objective To assess the diagnostic value of 18F-fluorodeoxyglucose (FDG) micro-PET/CT in evaluating an intracerebral hemorrhage (ICH) model in rats in vivo. Methods A simple random sampling method was used to select 32 healthy male adult SD rats, 4 of which were placed in the sham operation group and 28 rats were injected with 0.125 U/µL collagenase Ⅳ into the right basal ganglia to induce ICH (ICH model group). In the sham operation group, 0.9% saline was used instead of collagenase Ⅳ to make the sham model. The ICH model group was divided into seven groups by simple random sampling according to the time after ICH, which were 6, 24, 48 h and 3, 5, 7, 14 d (4 rats in each group). The sham operation and seven ICH model groups (6, 24, 48 h and 3, 5, 7, 14 d) underwent the neurological impairment scoring and 18F-FDG micro-PET/CT imaging. The hematoma volume at each time point in the ICH model group was calculated according to 18F-FDG micro-PET/CT imaging to delineate the region of interest (ROI) and Tada's formula. After imaging, the head was decapitated and the brain was obtained for hematoma observation and histopathological examination. The hematoma volume obtained by the two methods at the same time was compared by paired t-test and evaluated by Pearson correlation analysis. Results In the sham operation group: the neurological impairment scores were zero; and micro-PET/CT clearly showed homogeneous 18F-FDG uptake in the brain tissue. The ICH model group: the neurological impairment scores were (2.21±0.30), (3.51±0.66), (2.83±0.20), (2.12±0.50), (1.44±0.37), (1.02±0.25) and (0.51±0.12) at 6, 24, 48 h and 3, 5, 7, 14 d after ICH, respectively. At each time point, the 18F-FDG uptake decreased or became defective in the right basal ganglia of the rat brain. The cerebral hematoma volumes evaluated by 18F-FDG micro-PET/CT were (24.05±3.00), (27.19±1.25), (25.58±1.57), (21.94±0.98), (19.88±1.53), (18.35±2.11) and (16.29±1.53) mm3, respectively. The cerebral hematoma volumes evaluated by Tada's formula were (23.17±1.93), (26.09±1.35), (24.64±1.95), (21.31±1.32), (19.07±1.64), (17.29±1.38), and (15.63±1.98) mm3, respectively. No significant difference in the hematoma volume was found between two methods at each time point in the ICH model group (t=1.18−3.06, all P>0.05). The cerebral hematoma volume obtained by the two methods was significantly positively correlated (r=0.99, P<0.001). After the dissection of the ICH model group, irregular hematoma formation was seen in the right basal ganglia of the brain tissue, which was opposite to the radioactive sparse and defective areas shown by 18F-FDG micro-PET/CT. Conclusion 18F-FDG micro-PET/CT can accurately display the position, shape and size of the hematoma after ICH, and thus can be to verify the success of the rat ICH model in vivo. -
表 1 2种方法测得的脑出血模型组大鼠在脑出血后不同时间点脑血肿体积的比较[(
) mm3,n=4]$\bar{x}\pm s$ Table 1. Comparison of cerebral hematoma volume measured by two methods in rats in the intracerebral hemorrhage model group at different time points after intracerebral hemorrhage [(
)mm3,n=4]$\bar{x}\pm s $ 方法 6 h 24 h 48 h 3 d 5 d 7 d 14 d micro-PET/CT
勾画ROI法24.05±3.00 27.19±1.25 25.58±1.57 21.94±0.98 19.88±1.53 18.35±2.11 16.29±1.53 多田公式 23.17±1.93 26.09±1.35 24.64±1.95 21.31±1.32 19.07±1.64 17.29±1.38 15.63±1.98 t 值 1.58 1.18 3.06 1.80 2.21 2.84 1.91 P值 0.21 0.32 0.06 0.17 0.11 0.07 0.15 注:PET/CT为正电子发射断层显像计算机体层摄影术;多田公式为A×B×C/2(A和B分别为血肿最大层面测量的长径和短径;C为血肿切片数) -
[1] Jolink WMT, Wiegertjes K, Rinkel GJE, et al. Location-specific risk factors for intracerebral hemorrhage: systematic review and meta-analysis[J]. Neurology, 2020, 95(13): e1807−e1818. DOI: 10.1212/WNL.0000000000010418. [2] Kim JY, Bae HJ. Spontaneous intracerebral hemorrhage: management[J]. J Stroke, 2017, 19(1): 28−39. DOI: 10.5853/jos.2016.01935. [3] Majidi S, Olan WJ, Sigounas D. Intensive reduction of systolic blood pressure in acute intracerebral hemorrhage: is there a benefit?[J]. World Neurosurg, 2017, 101: 742−743. DOI: 10.1016/j.wneu.2017.03.079. [4] Cao HJ, Liu T, Li PF , et al. Altered long noncoding RNA and messenger RNA expression in experimental intracerebral hemorrhage—a preliminary study[J]. Cell Physiol Biochem, 2018, 45(3): 1284−1301. DOI: 10.1159/000487464. [5] Xi GH, Strahle J, Hua Y, et al. Progress in translational research on intracerebral hemorrhage: is there an end in sight?[J]. Prog Neurobiol, 2014, 115: 45−63. DOI: 10.1016/j.pneurobio.2013.09.007. [6] 杨凡慧, 曹龄之, 黄晓红, 等. 姜黄素治疗大鼠脑出血后脑葡萄糖代谢变化的18F-FDG microPET/CT显像观察[J]. 中华核医学与分子影像杂志, 2017, 37(10): 627−631. DOI: 10.3760/cma.j.issn.2095-2848.2017.10.006.
Yang FH, Cao LZ, Huang XH, et al. Protective effect of curcumin on glucose metabolism evaluated by 18F-FDG microPET/CT in rat models of intracerebral hemorrhage[J]. Chin J Nucl Med Mol Imaging, 2017, 37(10): 627−631. DOI: 10.3760/cma.j.issn.2095-2848.2017.10.006.[7] Shang HB, Cui DR, Yang DH, et al. The radical scavenger edaravone improves neurologic function and perihematomal glucose metabolism after acute intracerebral hemorrhage[J]. J Stroke Cerebrovasc Dis, 2015, 24(1): 215−222. DOI: 10.1016/j.jstrokecerebrovasdis.2014.08.021. [8] Takamatsu Y, Tamakoshi K, Waseda Y, et al. Running exercise enhances motor functional recovery with inhibition of dendritic regression in the motor cortex after collagenase-induced intracerebral hemorrhage in rats[J]. Behav Brain Res, 2016, 300: 56−64. DOI: 10.1016/j.bbr.2015.12.003. [9] 王雄, 邬玉芹, 陈红, 等. ApoE基因敲除构建大鼠缺血性脑卒中模型的实验研究[J]. 中国实验诊断学, 2020, 24(8): 1302−1306. DOI: 10.3969/j.issn.1007-4287.2020.08.024.
Wang X, Wu YQ, Chen H, et al. Experimental study of apoE gene knockout in rats with ischemic stroke model[J]. Chin J Lab Diagn, 2020, 24(8): 1302−1306. DOI: 10.3969/j.issn.1007-4287.2020.08.024.[10] Longa EZ, Weinstein PR, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20(1): 84−91. DOI: 10.1161/01.str.20.1.84. [11] Delcourt C, Carcel C, Zheng D, et al. Comparison of ABC methods with computerized estimates of intracerebral hemorrhage volume: the INTERACT2 study[J]. Cerebrovasc Dis Extra, 2019, 9(3): 148−154. DOI: 10.1159/000504531. [12] 徐兴华, 陈晓雷, 张军, 等. 多田公式计算脑内血肿体积的准确性和可靠性[J]. 中国神经精神疾病杂志, 2015, 41(2): 87−91. DOI: 10.3936/j.issn.1002-0152.2015.02.005.
Xu XH, Chen XL, Zhang J, et al. Study on the accuracy and reliability of the ABC/2 formula for volume assessment of intracerebral hematoma[J]. Chin J Nerv Ment Dis, 2015, 41(2): 87−91. DOI: 10.3936/j.issn.1002-0152.2015.02.005.[13] 邝忠华, 李成, 李兰君, 等. 高分辨率及高灵敏度小动物PET研究进展[J]. 原子核物理评论, 2016, 33(3): 336−344. DOI: 10.11804/NuclPhysRev.33.03.336.
Kuang ZH, Li C, Li LJ, et al. Progress of small animal PET scanners with high spatial resolution and high sensitivity[J]. Nucl Phys Rev, 2016, 33(3): 336−344. DOI: 10.11804/NuclPhysRev.33.03.336.[14] López-Gambero AJ, Martínez F, Salazar K, et al. Brain glucose-sensing mechanism and energy homeostasis[J]. Mol Neurobiol, 2019, 56(2): 769−796. DOI: 10.1007/s12035-018-1099-4. [15] 付怀栋, 潘永进, 王秀彬, 等. 磁共振成像在大鼠脑出血模型制作中的初步应用研究[J]. 重庆医学, 2012, 41(16): 1607−1608, 1611. DOI: 10.3969/j.issn.1671-8348.2012.16.017.
Fu HD, Pan YJ, Wang XB, et al. Preliminary study of MR imaging in making intracerebral hemorrhage model of rats[J]. Chongqing Med, 2012, 41(16): 1607−1608, 1611. DOI: 10.3969/j.issn.1671-8348.2012.16.017.[16] Zhu Y, Deng L, Tang HJ, et al. Electroacupuncture improves neurobehavioral function and brain injury in rat model of intracerebral hemorrhage[J]. Brain Res Bull, 2017, 131: 123−132. DOI: 10.1016/j.brainresbull.2017.04.003. [17] Wang Y, An FF, Chan M, et al. 18F-positron-emitting/fluorescent labeled erythrocytes allow imaging of internal hemorrhage in a murine intracranial hemorrhage model[J]. J Cereb Blood Flow Metab, 2017, 37(3): 776−786. DOI: 10.1177/0271678X16682510.