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蛛网膜下腔出血(SAH)是严重威胁人类生命的疾病, 病死率及致残率很高, 约40%的病人出血后48 h内死亡[1], 已广泛引起人们的关注与重视。近年来研究[2]显示, 改善大血管痉挛并不能明显改善SAH病人的预后, 早期脑损伤(EBI)可能是导致SAH病人死亡风险较高和决定预后的首要原因, 而且其发生机制复杂, 涉及脑细胞及脑血管一系列的病理生理改变。全脑CT灌注成像可以动态观察全脑不同时间段的微循环状况, 通过血流量(CBF)、血容量(CBV)、平均通过时间(MTT)和达峰时间(TTP)多参数定量分析, 间接反映EBI的进展程度。一氧化氮(NO)是重要的血管活性物质, 能够介导脑血管的舒缩反应, 影响EBI的进展。本研究拟通过兔SAH模型的建立, 研究EBI状态下脑微循环的变化规律及与血清NO的相关性。
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手术组术后20只兔表现为异常兴奋、上蹿下跳, 13只表现为淡漠、萎靡、活动减少, 4只出现活动障碍或瘫痪; 假手术组3只出现上述症状; 其余兔未见明显异常。
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手术组CBV与CBF在术后1、6、24、72 h均明显低于假手术组(P<0.01), MTT高于假手术组(P<0.05~P<0.01);手术组TTP在术后24 h和72 h高于假手术组间(P<0.01和P<0.05), 2组术后1 h和6 h差异无统计学意义(P>0.05)(见表 1)。
分组 n 术后1 h 术后6 h 术后24 h 术后72 h F P MS组内 CBV/(mL/100 g) 手术组 10 4.79±0.98 5.17±0.68 4.58±0.77 4.76±0.90 0.87 >0.05 0.706 假手术组 10 8.02±1.20 9.93±2.48* 10.95±1.05** 10.94±1.17** 7.56 < 0.01 2.516 t — 6.59 5.85 15.47 13.24 — — — P — < 0.01 < 0.01 < 0.01 < 0.01 — — — CBF/(mL·100 g-1·min-1) 手术组 10 353.71±42.94 257.32±37.25** 182.15±16.36**△△ 69.06±41.86**## 37.60 < 0.01 1 312.829 假手术组 10 632.45±95.69 657.52±61.15 559.14±104.30 550.79±115.29 3.04 < 0.05 9 266.543 t — 8.40* 17.67 11.29 12.42 — — — P — < 0.01 < 0.01 < 0.01 < 0.01 — — — MTT/s 手术组 10 1.76±0.21 2.19±0.43 2.70±0.67** 1.64±0.84## 6.67 < 0.01 0.346 假手术组 10 0.74±0.29 0.87±0.33 1.00±0.41 1.00±0.25 1.46 >0.05 0.106 t — 9.01 7.70 6.84 2.31 — — — P — < 0.01 < 0.01 < 0.01 < 0.05 — — — TTP/s 手术组 10 9.02±0.78 9.25±1.13 9.00±0.99 9.07±0.46 0.17 >0.05 0.769 假手术组 10 8.75±1.09 8.62±0.32*△ 7.73±0.58 8.31±0.71 3.87 < 0.05 0.533 t — 0.64 1.70 3.50 2.84 — — — P — >0.05 >0.05 < 0.01 < 0.05 — — — q检验: 与术后1 h比较*P<0.05, ** P<0.01;与术后6 h比较△△P<0.01;与术后24 h比较## P<0.01 表 1 2组CT灌注参数比较(x±s)
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假手术组在术后1、6、24、72 h NO含量无明显变化(P>0.05), 手术组兔在术后24 h和72 h NO含量有所回升(P<0.01);术后1、6、24、72 h手术组NO含量均明显低于相应假手术组(P<0.01)(见表 2)。
分组 n 术后1 h 术后6 h 术后24 h 术后72 h F P MS组内 手术组 10 0.66±0.15 0.84±0.21 1.93±0.55**△△ 1.85±0.50**△△ 28.41 < 0.01 0.155 假手术组 10 2.71±1.01 2.71±0.76 3.02±0.85 3.21±0.54 0.93 >0.05 0.653 t — 6.35 7.50 3.40 5.84 — — — P — < 0.01 < 0.01 < 0.01 < 0.01 — — — q检验: 与术后1 h比较** P<0.01;与术后6 h比较△△P<0.01 表 2 2组血清NO含量比较(x±s; μmol/L)
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直线相关分析显示, 手术组NO与MTT呈负相关关系(r=-0.854, P<0.05), 与CBF呈正相关关系(r=0.786, P<0.05), 与CBV(r=0.208, P>0.05)、TTP(r=0.311, P>0.05)间无明显相关性。
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大体观察: 术后1 h注血组, 枕大池区域、延髓腹侧面可见血液集聚, 大脑半球表面无明显血凝块。术后6 h注血组, 少量血液集聚于枕大池区域, 延髓腹侧面可见较多血凝块。术后24 h注血组, 枕大池区及大脑半球表面较多陈旧的血凝块。术后72 h注血组, 枕大池内未见明显的血凝块, 而大脑半球腹侧面可见少量血凝块。假手术组大脑半球表面无明显血凝块(见图 2)。
镜下观察: 1 h手术组, 颞叶及枕叶部分小血管略狭窄, 血管周围出现少量炎性细胞的浸润; 6 h手术组, 额、颞、枕叶出现较明显炎症反应, 血管管腔狭窄, 管壁增厚, 周围淋巴细胞增多聚集; 24 h手术组, 额、顶叶局部出现无结构的均质红染的坏死区, 周围有大量炎性细胞浸润, 颞、枕叶小血管明显狭窄, 细胞肿胀; 72 h手术组, 额、枕、颞叶脑组织细胞增大, 水肿, 胞质增多。假手术组病理切片示脑组织结构清晰, 细胞大小形态分布正常, 未见明显炎症反应和坏死灶(见图 3)。
一氧化氮与蛛网膜下腔出血后早期脑损伤所致脑微循环变化的相关性研究
Correlation between nitric oxide and cerebral microcirculation changes caused by early brain injury after subarachnoid hemorrhage
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摘要:
目的观察蛛网膜下腔出血(SAH)后早期脑微循环的变化及SAH后一氧化氮(NO)的表达对早期脑损伤(EBI)及脑微循环的影响。 方法80只新西兰大白兔分为手术组和假手术组,2组再分为术后1、6、24、72 h 4个亚组,各10只。手术组采用枕大池注血法制备兔SAH模型,假手术组采用相同方法注射0.9%氯化钠溶液制备模型。然后行全脑CT灌注成像,比较各组血流量(CBF)、血容量(CBV)、平均通过时间(MTT)、达峰时间(TTP)、NO含量,并分析NO与CBV、CBF、MTT、TTP相关性。 结果手术组CBV与CBF在术后1、6、24、72 h均明显低于假手术组(P<0.01),MTT高于假手术组(P<0.05~P<0.01);手术组TTP在术后24 h和72 h均高于假手术组间(P<0.01和P<0.05)。假手术组在术后1、6、24、72 h NO含量无明显变化(P>0.05),手术组兔在术后24 h和72 h NO含量有所回升(P<0.01);术后1、6、24、72 h手术组NO含量均明显低于相应假手术组(P<0.01)。直线相关分析显示,手术组NO与MTT呈负相关关系(r=-0.854,P<0.05),与CBF呈正相关关系(r=0.786,P<0.05)。 结论CT灌注能早期发现SAH后局部脑缺血,反映SAH后EBI的病理进展程度。NO一定程度上影响SAH后微循环的改变及EBI的进展程度。 Abstract:ObjectiveTo observe the changes of early cerebral microcirculation after subarachnoid hemorrhage (SAH) and the effect of nitric oxide (NO) expression on early brain injury (EBI) and cerebral microcirculation after SAH. MethodsEighty New Zealand white rabbits were divided into the operation group and sham operation group, then the two groups were further divided into 4 subgroups including 1 h, 6 h, 24 h and 72 h after operation, with 10 rabbits in each subgroup.SAH model was established by injecting autologous blood into cisterna magna in operation group, and injecting 0.9% sodium chloride solution with the same method in sham operation group.The CT perfusion imaging of whole brain was performed to compare the cerebral blood flow(CBF), cerebral blood volume(CBV), mean transit time(MTT), time to peak (TTP) and NO content in each group, and the correlation between NO and CBV, CBF, MTT, TTP was analyzed. ResultsCBV and CBF in operation group were significantly lower than those in sham operation group at 1 h, 6 h, 24 h and 72 h after operation (P<0.01), and MTT was higher than that in sham operation group(P<0.05 to P<0.01);TTP in operation group was higher than that in sham operation group at 24 h and 72 h after operation(P<0.01 and P<0.05).There was no significant change in the NO content at 1 h, 6 h, 24 h and 72 h after operation in sham operation group(P>0.05), but the NO content increased at 24 h and 72 h after operation in operation group(P<0.01);at 1 h, 6 h, 24 h and 72 h after operation, the NO content in operation group was significantly lower than that in the corresponding sham operation group(P<0.01).Linear correlation analysis showed that NO was negatively correlated with MTT(r=-0.854, P<0.05) and positively correlated with CBF(r=0.786, P<0.05). ConclusionsCT perfusion can detect the focal cerebral ischemia after SAH in early stage, which reflects the pathological progress of EBI after SAH.NO affects the microcirculation changes and the progress of EBI after SAH to some extent. -
Key words:
- subarachnoid hemorrhage /
- early brain injury /
- nitric oxide /
- CT perfusion /
- microcirculation
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表 1 2组CT灌注参数比较(x±s)
分组 n 术后1 h 术后6 h 术后24 h 术后72 h F P MS组内 CBV/(mL/100 g) 手术组 10 4.79±0.98 5.17±0.68 4.58±0.77 4.76±0.90 0.87 >0.05 0.706 假手术组 10 8.02±1.20 9.93±2.48* 10.95±1.05** 10.94±1.17** 7.56 < 0.01 2.516 t — 6.59 5.85 15.47 13.24 — — — P — < 0.01 < 0.01 < 0.01 < 0.01 — — — CBF/(mL·100 g-1·min-1) 手术组 10 353.71±42.94 257.32±37.25** 182.15±16.36**△△ 69.06±41.86**## 37.60 < 0.01 1 312.829 假手术组 10 632.45±95.69 657.52±61.15 559.14±104.30 550.79±115.29 3.04 < 0.05 9 266.543 t — 8.40* 17.67 11.29 12.42 — — — P — < 0.01 < 0.01 < 0.01 < 0.01 — — — MTT/s 手术组 10 1.76±0.21 2.19±0.43 2.70±0.67** 1.64±0.84## 6.67 < 0.01 0.346 假手术组 10 0.74±0.29 0.87±0.33 1.00±0.41 1.00±0.25 1.46 >0.05 0.106 t — 9.01 7.70 6.84 2.31 — — — P — < 0.01 < 0.01 < 0.01 < 0.05 — — — TTP/s 手术组 10 9.02±0.78 9.25±1.13 9.00±0.99 9.07±0.46 0.17 >0.05 0.769 假手术组 10 8.75±1.09 8.62±0.32*△ 7.73±0.58 8.31±0.71 3.87 < 0.05 0.533 t — 0.64 1.70 3.50 2.84 — — — P — >0.05 >0.05 < 0.01 < 0.05 — — — q检验: 与术后1 h比较*P<0.05, ** P<0.01;与术后6 h比较△△P<0.01;与术后24 h比较## P<0.01 表 2 2组血清NO含量比较(x±s; μmol/L)
分组 n 术后1 h 术后6 h 术后24 h 术后72 h F P MS组内 手术组 10 0.66±0.15 0.84±0.21 1.93±0.55**△△ 1.85±0.50**△△ 28.41 < 0.01 0.155 假手术组 10 2.71±1.01 2.71±0.76 3.02±0.85 3.21±0.54 0.93 >0.05 0.653 t — 6.35 7.50 3.40 5.84 — — — P — < 0.01 < 0.01 < 0.01 < 0.01 — — — q检验: 与术后1 h比较** P<0.01;与术后6 h比较△△P<0.01 -
[1] 张云辉, 王廷华. 脑源性神经营养因子与脑缺血肺水肿[J]. 四川大学学报(医学版), 2012, 43(6): 893. [2] LIU E, SUN L, ZHANG Y, et al. Aquaporin4 knockout aggravates early brain injury following subarachnoid hemorrhage through impairment of the glymphatic system in rat brain[J]. Acta neurochirurgica Supplement, 2020, 127: 59. [3] GU H, FEI ZH, WANG YQ, et al. Angiopoietin-1 and angiopoiein-2 expression imbalance influence in early period after subarachnoid hemorrhage[J]. Int Neurourol J, 2016, 20(4): 288. doi: 10.5213/inj.1632692.346 [4] GÜRER B, KERTMEN H, KURU BEKTAŞOǦLU P, et al. The effects of cinnamaldehyde on early brain injury and cerebral vasospasm following experimental subarachnoid hemorrhage in rabbits[J]. Metab Brain Dis, 2019, 34(6): 1737. doi: 10.1007/s11011-019-00480-7 [5] ZHANG H, ZHANG B, LI S, et al. Whole brain CT perfusion combined with CT angiography in patients with subarachnoid hemorrhage and cerebral vasospasm[J]. Clin Neurol Neurosurg, 2013, 115(12): 2496. doi: 10.1016/j.clineuro.2013.10.004 [6] VAIDYA T, AGRAWAL A, MAHAJAN S, et al. The continuing evolution of molecular functional imaging in clinical oncology: The road to precision medicine and radiogenomics(Part Ⅰ)[J]. Mol Diagn Ther, 2019, 23(1): 1. doi: 10.1007/s40291-018-0366-4 [7] OEBEL S, HAMADA S, HIGASHIGAITO K, et al. Comprehensive morphologic and functional imaging of heart transplant patients: first experience with dynamic perfusion CT[J]. Eur Radiol, 2018, 28(10): 4111. doi: 10.1007/s00330-018-5436-9 [8] QIN L, LI S, ZHENG RB, et al. Endothelin-1 expression and alterations of cerebral microcirculation after experimental subarachnoid hemorrhage[J]. Neuroradiology, 2015, 57(1): 63. doi: 10.1007/s00234-014-1435-y [9] MALINOVA V, ILIEV B, TSOGKAS I, et al. Assessment of tissue permeability by early CT perfusion as a surrogate parameter for early brain injury after subarachnoid hemorrhage[J]. J Neurosurg, 2019, 23: 1. [10] CHAI WN, SUN XC, LV FJ, et al. Clinical study of changes of cerebral microcirculation in cerebral vasospasm after SAH[J]. Acta Neurochir Suppl, 2011, 110(1): 225. [11] WANG Z, SHI XY, YIN J, et al. Role of autophary in early brain injury after experimental subarachnoid hemorrhage[J]. J Mol Neurosci, 2011, 28(6): 252. [12] OKADA T, ENKHJARGAL B, TRAVIS ZD, et al. FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage[J]. Mol Neurobiol, 2019, 56(12): 8203. doi: 10.1007/s12035-019-01668-9 [13] ZHOU N, XU T, BAI Y, et al. Protective effects of urinary trypsin inhibitor on vascular permeability following subarachnoid hemorrhage in a rat model[J]. CNS Neurosci Ther, 2013, 19(9): 659. doi: 10.1111/cns.12122 [14] ZHENG RB, QIN L, LI SB, et al. CT perfusion-derived mean transit time of cortical brain has a negative correlation with the plasma level of Nitric Oxide after subarachnoid hemorrhage[J]. Acta Neurochir, 2014, 156(3): 527. doi: 10.1007/s00701-013-1968-6 [15] KIM H, BRITTON GL, PENG T, et al. Nitric oxide-loaded echogenic liposomes for treatment of vasospasm following subarachnoid hemorrhage[J]. Int J Nanomedicine, 2014, 9: 155. [16] ZHAO D, LIU Q, JI Y, et al. Correlation between nitric oxide and early brain injury after subarachnoid hemorrhage[J]. Int J Neurosci, 2015, 37(6): 476. [17] WU Q, ZHENG R, WANG J, et al. CT perfusion imaging of cerebral microcirculatory changes following subarachnoid hemorrhage in rabbits: Specific role of endothelin-1 receptor antagonist[J]. Brain Res, 2018, 12: 1701. [18] LI HT, WANG J, LI SF, et al. Upregulation of microRNA-24 causes vasospasm following subarachnoid hemorrhage by suppressing the expression of endothelial nitric oxide synthase[J]. Mol Med Rep, 2018, 18(1): 1181. [19] SUN J, ZHANG Y, LU J, et al. Salvinorin A ameliorates cerebral vasospasm through activation of endothelial nitric oxide synthase in a rat model of subarachnoid hemorrhage[J]. Microcirculation, 2018, 25(3): e12442. doi: 10.1111/micc.12442