Comparative assessment of in vitro BBB tight junction integrity following exposure to cigarette smoke and e-cigarette vapor: a quantitative evaluation of the protective effects of metformin using small-molecular-weight paracellular markers.
Comparative assessment of in vitro BBB tight junction integrity following exposure to cigarette smoke and e-cigarette vapor: a quantitative evaluation of the protective effects of metformin using small-molecular-weight paracellular markers.
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DOI：
10.1186/s12987-021-00261-4
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发表时间：
2021-06-22
影响因子：
7.3
通讯作者：
Cucullo L
中科院分区：
文献类型：
--
作者：
Kadry H;Noorani B;Bickel U;Abbruscato TJ;Cucullo L
血脑屏障（BBB）在保护中枢神经系统（CNS）免受血源性物质和潜在有害的外源性物质的影响方面起着关键作用。我们小组以前的数据表明，烟草烟雾（TS）和电子烟（EC）影响BBB的完整性，增加中风的发生率，并被认为是多种CNS疾病的危险因素。还发现二甲双胍可消除TS和EC的不良影响。我们使用蔗糖和甘露醇作为细胞旁标记物来定量评估TS和EC对体外血脑屏障的影响。具体而言，我们使用定量平台来确定吸烟对BBB的有害影响，并研究二甲双胍的保护作用。使用transwell系统和iPSC衍生的BMEC，我们评估了TS和EC在不同时间点在有和没有二甲双胍预处理的情况下对蔗糖和甘露醇渗透性的影响。同时，使用免疫荧光（IF）和蛋白质印迹（WB）技术，我们评估了紧密连接蛋白的表达和分布，包括ZO-1，occludin和claudin-5。我们的数据表明，TS和EC对蔗糖和甘露醇的渗透性产生负面影响，从6小时后开始，直到24小时。屏障完整性的丧失与TEER值的降低相关。虽然ZO-1和occludin的总体表达水平没有显著下调，但ZO-1的分布发生了改变，通过IF成像可以明显看出停药模式。与occludin相反，紧密连接蛋白-5的表达显著降低TS和EC，如WB和IF数据所示。与先前的研究一致，我们的数据显示二甲双胍可以抵消TS和EC对BBB完整性的负面影响，从而表明重新利用这种药物来提供脑血管保护的可能性。
The blood–brain barrier (BBB) plays a critical role in protecting the central nervous system (CNS) from blood-borne agents and potentially harmful xenobiotics. Our group’s previous data has shown that tobacco smoke (TS) and electronic cigarettes (EC) affect the BBB integrity, increase stroke incidence, and are considered a risk factor for multiple CNS disorders. Metformin was also found to abrogate the adverse effects of TS and EC. We used sucrose and mannitol as paracellular markers to quantitatively assess TS and EC’s impact on the BBB in-vitro. Specifically, we used a quantitative platform to determine the harmful effects of smoking on the BBB and study the protective effect of metformin. Using a transwell system and iPSCs-derived BMECs, we assessed TS and EC’s effect on sucrose and mannitol permeability with and without metformin pre-treatment at different time points. Concurrently, using immunofluorescence (IF) and Western blot (WB) techniques, we evaluated the expression and distribution of tight junction proteins, including ZO-1, occludin, and claudin-5. Our data showed that TS and EC negatively affect sucrose and mannitol permeability starting after 6 h and up to 24 h. The loss of barrier integrity was associated with a reduction of TEER values. While the overall expression level of ZO-1 and occludin was not significantly downregulated, the distribution of ZO-1 was altered, and discontinuation patterns were evident through IF imaging. In contrast to occludin, claudin-5 expression was significantly decreased by TS and EC, as demonstrated by WB and IF data. In agreement with previous studies, our data showed the metformin could counteract the negative impact of TS and EC on BBB integrity, thus suggesting the possibility of repurposing this drug to afford cerebrovascular protection.
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影响因子：
3.4
作者：
Das, Subhasis;Gautam, N.;Roy, Somenath
通讯作者：
Roy, Somenath
影响因子：
--
作者：
GILL, JS;SHIPLEY, MJ;BEEVERS, DG
通讯作者：
BEEVERS, DG
影响因子：
5.3
作者：
Bhowmick, Saurav;D'Mello, Veera;Abdul-Muneer, P. M.
通讯作者：
Abdul-Muneer, P. M.
DOI：
10.1016/j.jstrokecerebrovasdis.2013.09.001
发表时间：
2014-02-01
影响因子：
2.5
作者：
Cheng, Yuan-Yang;Leu, Hsin-Bang;Kao, Chung-Lan
通讯作者：
Kao, Chung-Lan
影响因子：
5
作者：
Boven, LA;Middel, J;Nottet, HSLM
通讯作者：
Nottet, HSLM