Department of Neurology, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Neurology, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China; Hebei Key Laboratory of Vascular Homeostasis and Hebei Collaborative Innovation Center for Cardio-cerebrovascular Diseases, Shijiazhuang, Hebei, China.
Exp Neurol. 2018 Apr;302:68-74. doi: 10.1016/j.expneurol.2017.12.016. Epub 2017 Dec 30.
The effects of Ulinastatin (UTI) on the blood-brain barrier (BBB) in the acute phase of cerebral ischemia/reperfusion (I/R) are not clear. This study was to investigate the potential protective effects of UTI on the BBB and the underlying mechanisms.
Male CD-1 mice were subjected to transient middle cerebral artery occlusion (tMCAO) and randomly assigned to four groups: Sham (sham-operated), tMCAO (tMCAO+0.9% saline), UTI-L (tMCAO+UTI 1500U/100g) and UTI-H (tMCAO+UTI 3000U/100g) group. UTI was administered immediately after reperfusion in the UTI-L and UTI-H groups. At 24h after reperfusion, the neurological deficit, brain water content, and infarct volume were determined. Western blot and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were used to examine the expression of matrix metalloproteinase (MMP)-9, Zonula occludens-1 (ZO-1) and occludin in ischemic cerebral cortex. The integrity of the BBB was assessed by the leakage of Evans blue.
Compared with tMCAO group, both UTI-L and UTI-H groups showed significantly (P<0.001) ameliorated the neurological deficit (2.00±0.71 and 1.60±0.55 vs. 4.60±0.55), lessened brain water content (82.99%±0.21% and 82.05%±0.59% vs. 84.28%±0.0.57%) and decreased the infarct volume (38.52%±1.72% and 24.78%±1.20% vs. 49.48%±1.93%). In addition, significantly (P<0.001) decreased expression of MMP-9 (0.48±0.06 and 0.37±0.05 vs.0.76±0.10 for protein and 2.88±0.23 and 2.17±0.16 vs. 3.90±0.24 for mRNA) and alleviated loss of ZO-1 (0.19±0.04 and 0.24±0.05 vs. 0.25±0.03) and occludin (0.74±0.08 and 0.87±0.07 vs. 0.94±0.06) proteins were observed in both UTI-L and UTI-H groups.
UTI protects the brain against ischemic injury potentially via down-regulating the expression of MMP-9 and alleviating loss of ZO-1 and occludin proteins to restore the BBB permeability.
尿胰蛋白酶抑制剂（UTI）对脑缺血再灌注（I/R）急性期血脑屏障（BBB）的影响尚不清楚。本研究旨在探讨 UTI 对 BBB 的潜在保护作用及其机制。
雄性 CD-1 小鼠进行短暂性大脑中动脉闭塞（tMCAO），并随机分为四组：假手术组（Sham）、tMCAO 组（tMCAO+0.9%生理盐水）、UTI-L 组（tMCAO+UTI 1500U/100g）和 UTI-H 组（tMCAO+UTI 3000U/100g）。UTI-L 和 UTI-H 组在再灌注后立即给予 UTI。再灌注 24h 后，测定神经功能缺损、脑水含量和梗死体积。Western blot 和定量逆转录聚合酶链反应（qRT-PCR）检测缺血皮质中基质金属蛋白酶（MMP）-9、紧密连接蛋白-1（ZO-1）和闭合蛋白的表达。伊文思蓝渗漏评估 BBB 的完整性。
与 tMCAO 组相比，UTI-L 和 UTI-H 组的神经功能缺损均明显改善（2.00±0.71 和 1.60±0.55 比 4.60±0.55），脑水含量降低（82.99%±0.21% 和 82.05%±0.59% 比 84.28%±0.0.57%），梗死体积减小（38.52%±1.72% 和 24.78%±1.20% 比 49.48%±1.93%）。此外，MMP-9 的表达明显降低（蛋白 0.48±0.06 和 0.37±0.05 比 0.76±0.10，mRNA 2.88±0.23 和 2.17±0.16 比 3.90±0.24），ZO-1（蛋白 0.19±0.04 和 0.24±0.05 比 0.25±0.03）和闭合蛋白（蛋白 0.74±0.08 和 0.87±0.07 比 0.94±0.06）的丢失也明显减轻。
UTI 通过下调 MMP-9 的表达和减轻 ZO-1 和闭合蛋白的丢失来恢复 BBB 的通透性，从而对缺血性损伤起到保护作用。
