Iwao Takuro, Takata Fuyuko, Aridome Hisataka, Yasunaga Miho, Yokoya Miki, Mizoguchi Junko, Dohgu Shinya
Department of Pharmaceutical Care and Health Sciences, Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan.
Int J Mol Sci. 2025 Jun 20;26(13):5949. doi: 10.3390/ijms26135949.
The brain actively obtains nutrients through various transporters on brain microvessel endothelial cells (BMECs). Major facilitator superfamily domain-containing protein 2a (MFSD2A) serves as a key transporter of docosahexaenoic acid (DHA) at the blood-brain barrier (BBB) and is exclusively expressed in BMECs. Although brain pericytes (PCs) regulate MFSD2A expression in BMECs, the underlying mechanism remains unclear. To determine whether PDGF-BB/PDGFRβ signaling between endothelial cells (ECs) and PCs affects MFSD2A protein expression and plasma membrane localization in ECs, we examined the impact of AG1296 (a PDGF receptor inhibitor) and -knockdown PCs on a non-contact coculture BBB model comprising the primary cultures of rat brain ECs and PCs. The effects of PCs on MFSD2A expression, localization, and brain endothelial DHA uptake was assessed using Western blot, immunofluorescence staining, and [C]DHA uptake by ECs, respectively. In ECs cocultured with PCs, MFSD2A expression and plasma membrane localization were significantly higher than in EC monolayers. Moreover, conditioned medium derived from PCs failed to enhance MFSD2A expression. The increased expression and membrane localization of MFSD2A were inhibited by AG1296 and -knockdown PCs. Furthermore, PCs significantly increased [C]DHA uptake by ECs. These findings suggest that PCs enhance MFSD2A expression and plasma membrane localization in ECs through PDGF-BB/PDGFRβ signaling.
大脑通过脑微血管内皮细胞（BMECs）上的各种转运蛋白积极获取营养物质。含主要易化子超家族结构域蛋白2a（MFSD2A）是血脑屏障（BBB）处二十二碳六烯酸（DHA）的关键转运蛋白，且仅在BMECs中表达。尽管脑周细胞（PCs）调节BMECs中MFSD2A的表达，但其潜在机制仍不清楚。为了确定内皮细胞（ECs）和PCs之间的血小板源性生长因子-BB/血小板源性生长因子受体β（PDGF-BB/PDGFRβ）信号是否影响ECs中MFSD2A蛋白表达和质膜定位，我们在由大鼠脑ECs和PCs原代培养物组成的非接触共培养血脑屏障模型中检测了AG1296（一种PDGF受体抑制剂）和PCs基因敲低的影响。分别使用蛋白质免疫印迹法、免疫荧光染色法以及ECs对[C]DHA的摄取来评估PCs对MFSD2A表达、定位和脑内皮细胞DHA摄取的影响。在与PCs共培养的ECs中，MFSD2A的表达和质膜定位显著高于ECs单层。此外，来自PCs的条件培养基未能增强MFSD2A的表达。AG1296和PCs基因敲低抑制了MFSD2A表达和膜定位的增加。此外，PCs显著增加了ECs对[C]DHA的摄取。这些发现表明，PCs通过PDGF-BB/PDGFRβ信号增强ECs中MFSD2A的表达和质膜定位。
