首页> 外文期刊>American Journal of Physiology >Carbonic anhydrase IX is a critical determinant of pulmonary microvascular endothelial cell pH regulation and angiogenesis during acidosis
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Carbonic anhydrase IX is a critical determinant of pulmonary microvascular endothelial cell pH regulation and angiogenesis during acidosis

机译:碳酸酐酶IX是妇女中毒期间肺部微血管内皮细胞pH调节和血管生成的关键决定因素

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Carbonic anhydrase IX (CA IX) is highly expressed in rapidly proliferating and highly glycolytic cells, where it serves to enhance acid-regulatory capacity. Pulmonary microvascular endothelial cells (PMVECs) actively utilize aerobic glycolysis and acidify media, whereas pulmonary arterial endothelial cells (PAECs) primarily rely on oxidative phosphorylation and minimally change media pH. Therefore, we hypothesized that CA IX is critical to PMVEC angiogenesis because of its important role in regulating pH. To test this hypothesis, PMVECs and PAECs were isolated from Sprague-Dawley rats. CA IX knockout PMVECs were generated using the CRISPR-Cas9 technique. During serum-stimulated growth, mild acidosis (pH 6.8) did not affect cell counts of PMVECs, but it decreased PAEC cell number. Severe acidosis (pH 6.2) decreased cell counts of PMVECs and elicited an even more pronounced reduction of PAECs. PMVECs had a higher CA IX expression compared with PAECs. CA activity was higher in PMVECs compared with PAECs, and enzyme activity was dependent on the type IX isoform. Pharmacological inhibition and genetic ablation of CA IX caused profound dysregulation of extra- and intracellular pH in PMVECs. Matrigel assays revealed impaired angiogenesis of CA IX knockout PMVECs in acidosis. Lastly, pharmacological CA IX inhibition caused profound cell death in PMVECs, whereas genetic CA IX ablation had little effect on PMVEC cell death in acidosis. Thus CA IX controls PMVEC pH necessary for angiogenesis during acidosis. CA IX may contribute to lung vascular repair during acute lung injury that is accompanied by acidosis within the microenvironment.
机译:碳酸酐酶IX(CaI1)在快速增殖和高糖酵解细胞中高度表达,在那里它用于提高酸性调节能力。肺部微血管内皮细胞(PMVEC)积极地利用有氧糖醇分解和酸化培养基,而肺动脉内皮细胞(PAEC)主要依赖于氧化磷酸化和微小变化培养基pH。因此,我们假设Ca Ix对PMVEC血管生成至关重要,因为其在调节pH中的重要作用。为了测试这个假设,从Sprague-Dawley大鼠分离PMVEC和PAEC。使用CRISPR-CAS9技术产生CA IX敲除PMVEC。在血清刺激的生长期间,轻度酸中毒(pH6.8)不影响PMVEC的细胞计数,但它降低了PAEC细胞数。严重酸中毒(pH6.2)降低了PMVEC的细胞计数,并引发了更明显的PAEC减少。与PAEC相比,PMVecs具有更高的CA IX表达式。与PAEC相比,PMVEC的Ca活性较高,酶活性取决于IX型同种型。 CAIX的药理抑制和遗传烧蚀引起了PMVEC中的高度和细胞内pH的深远困难。 Matrigel测定揭示了Ca IX敲除PMVECs在酸中的血管生成受损。最后,药理CA IX抑制在PMVEC中引起了深刻的细胞死亡,而遗传CAIX消融对PMVEC细胞死亡对酸中毒的影响几乎没有影响。因此,CA IX控制酸中毒期间血管生成所需的PMVEC pH。 CA IX可能有助于急性肺损伤期间的肺血管修复,伴随着微环境内的酸中毒。

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