Ulinastatin protects brain against cerebral ischemia/reperfusion injury through inhibiting MMP-9 and alleviating loss of ZO-1 and occludin proteins in mice
Ulinastatin protects brain against cerebral ischemia/reperfusion injury through inhibiting MMP-9 and alleviating loss of ZO-1 and occludin proteins in mice
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DOI：
10.1016/j.expneurol.2017.12.016
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发表时间：
2018-04-01
影响因子：
5.3
通讯作者：
Gao, Bu-Lang
中科院分区：
文献类型：
--
作者：
Li, Xiao-Fang;Zhang, Xiang-Jian;Gao, Bu-Lang
背景：乌司他丁(Ulinastatin，UTI)对脑缺血再灌流(I/R)急性期血脑屏障(BBB)的影响尚不清楚。方法：雄性CD-1小鼠建立短暂性大脑中动脉闭塞(TMCAO)模型，随机分为假手术组、tMCAO+0.9%生理盐水组、UTI-L组(tMCAO+UTI1500U/100g)和UTI-H组(tMCAO+UTI3000U/100g)。UTI-L组和UTI-H组在再灌流后即刻给予UTI。再灌流后24小时测定神经功能、脑含水量和脑梗塞体积。采用Western印迹和定量逆转录聚合酶链式反应(qRT-PCR)检测缺血大脑皮层基质金属蛋白酶-9(MMP-9)、闭锁小带-1(ZO-1)和闭锁蛋白的表达。通过伊文思蓝渗漏来评估血脑屏障的完整性。结果：与TMCAO组比较，UTI-L组和UTI-H组均有显著差异(P<；0.001)可改善神经功能缺失(2.00±0.71和1.60±0.55 vs 4.60±0.55)，减少脑含水量(82.99%±0.21%和82.05%±0.59%vs 84.28%±0.57%)，缩小脑梗塞体积(38.52%±1.72%和24.78%±1.20%vs 49.48%+/-1.93%)。此外，显著(P<；0.001)减少MMP9的表达(0.480.06和0.37+/-0.05vs.0.76+/-0.10mRNA：2.88+/-0.23和2.17+/-0.16vs.3.90+/-0.24)，减轻ZO-1(0.19+/-0.04和0.240.05：0.25+/-0.03)和occludin(0.74+/-0)的丢失0.08和0.87+/-0.07vs.0.94+/-0.06)。结论：UTI可能通过下调MMP9的表达，减轻ZO-1和occludin蛋白的丢失，恢复血脑屏障通透性，对脑缺血损伤具有保护作用。
Background: 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.& para;Methods: 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 1500 U/100 g) and UTI-H (tMCAO + UTI 3000 U/100 g) group. UTI was administered immediately after reperfusion in the UTI-L and UTI-H groups. At 24 h 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. & para;& para;Results: 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.& para;& para;Conclusion: 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.