脑淀粉样血管病 （CAA）（Cerebral amyloid angiopathy：CAA） 是老年人认知障碍和脑出血 （ICH） 的主要原因。纤维状淀粉样蛋白β蛋白（Aβ）在小血管和毛细血管内的沉积，尤其是β淀粉样蛋白肽1-40（Aβ40）是CAA的特征。巨噬细胞谱系细胞的吞噬行为，包括卵黄囊衍生的小胶质细胞（Microglia），脑驻留血管周围巨噬细胞（PeriVascular Macrophages：PVM）和循环单核细胞（Ly6Chi）衍生的浸润巨噬细胞，对于Aβ清除至关重要。巨噬细胞清除活性受损已被证明会加剧CAA结果。另一方面，发现被吞噬货物淹没的巨噬细胞谱系细胞获得了组织破坏特性。载有Aβ的巨噬细胞在CAA中的病理影响有待研究。

为探讨三种巨噬细胞在脑淀粉样血管病 （CAA）的功能，研究者使用了荷兰Liposoma的巨噬细胞清除剂Clodronate liposomes氯膦酸盐脂质体和饮食给药的PLX5622。外周血循环单核细胞以10ul/g剂量通过腹腔注射来清除；小胶质细胞通过长期饮食喂养PLX5622来清除；脑驻留血管周围巨噬细胞通过脑立体定位注射10ul荷兰Liposoma的巨噬细胞清除剂Clodronate liposomes氯膦酸盐脂质体来清除。三种巨噬细胞的清除效果通过流式或者免疫荧光来评价。

Peripheral blood monocyte depletion was carried out according to published procedure35. Clodronate liposomes (Liposoma, 10?ml/kg, i.p.) was administered to 12-week-old Tg-SwDI/B mice every 3 days to deplete peripheral monocytes/macrophages prior to sacrifice at day 7. Depletion efficacy was confirmed with flow cytometric analysis.

For microglia depletion, PLX5622 was supplied to 12-week-old Tg-SwDI/B mice in the diet at 1200 PPM (1200?mg/kg of chow), starting 7 days prior to sacrifice36. Depletion efficacy of was confirmed with immunostaining.

Perivascular macrophage depletion was carried out according to published procedure9. 12-week-old Tg-SwDI/B mice were anesthetized with isoflurane and stereotaxically injected with 10?μL of clodronate-containing liposomes into the left lateral ventricle (coordinates from Bregma: anteroposterior, 0.2?mm; mediolateral, 1.2?mm; dorsoventral, 2.3?mm). Animals were sacrificed 7 days later and brains were processed for further analysis. Depletion efficacy of was confirmed with immunostaining.

清除效果：

CD206 (green, PVM marker），见图a

F4/80 (green, infiltrated macrophages marker，见图b

Clo ICV：intra-cerebral ventricle injected clodronate liposomes (Clo ICV) ，脑室内注射clodronate liposomes

CLO IP：intraperitoneally injected clodronate liposomes (CLO IP)，腹腔注射clodronate liposomes

见图C：CD45和F4/80双标的流式图，外周血循环单核细胞以10ul/g剂量通过腹腔注射来清除，相对于对照的6.7%，荷兰Liposoma的巨噬细胞清除剂Clodronate liposomes氯膦酸盐脂质体可以清除94%以上的外周血单核细胞巨噬细胞。见图d：小胶质细胞通过长期饮食喂养PLX5622来清除；见图e：脑驻留血管周围巨噬细胞通过脑立体定位注射10ul荷兰Liposoma的巨噬细胞清除剂Clodronate liposomes氯膦酸盐脂质体来清除。

论文信息：

论文题目：Macrophage lineage cells-derived migrasomes activate complement-dependent blood-brain barrier damage in cerebral amyloid angiopathy mouse model

期刊名称：Nature Communications

时间期卷：14, Article number: 3945 (2023)

在线时间：2023年7月4日

DOI：doi.org/10.1038/s41467-023-39693-x

产品信息：

货号：CP-005-005

规格：5ml+5ml

品牌：Liposoma

产地：荷兰

名称：Clodronate Liposomes and Control Liposomes

办事处：Target Technology(靶点科技)