氯膦酸鹽脂質(zhì)體和PLX5622清除腦部三種巨噬細(xì)胞解決方案

腦淀粉樣血管病 （CAA）（Cerebral amyloid angiopathy：CAA） 是老年人認(rèn)知障礙和腦出血 （ICH） 的主要原因。纖維狀淀粉樣蛋白β蛋白（Aβ）在小血管和毛細(xì)血管內(nèi)的沉積，尤其是β淀粉樣蛋白肽1-40（Aβ40）是CAA的特征。巨噬細(xì)胞譜系細(xì)胞的吞噬行為，包括卵黃囊衍生的小膠質(zhì)細(xì)胞（Microglia），腦駐留血管周圍巨噬細(xì)胞（PeriVascular Macrophages：PVM）和循環(huán)單核細(xì)胞（Ly6Chi）衍生的浸潤巨噬細(xì)胞，對于Aβ清除至關(guān)重要。巨噬細(xì)胞清除活性受損已被證明會加劇CAA結(jié)果。另一方面，發(fā)現(xiàn)被吞噬貨物淹沒的巨噬細(xì)胞譜系細(xì)胞獲得了組織破壞特性。載有Aβ的巨噬細(xì)胞在CAA中的病理影響有待研究。

為探討三種巨噬細(xì)胞在腦淀粉樣血管病 （CAA）的功能，研究者使用了荷蘭Liposoma的巨噬細(xì)胞清除劑Clodronate liposomes氯膦酸鹽脂質(zhì)體和飲食給藥的PLX5622。外周血循環(huán)單核細(xì)胞以10ul/g劑量通過腹腔注射來清除；小膠質(zhì)細(xì)胞通過長期飲食喂養(yǎng)PLX5622來清除；腦駐留血管周圍巨噬細(xì)胞通過腦立體定位注射10ul荷蘭Liposoma的巨噬細(xì)胞清除劑Clodronate liposomes氯膦酸鹽脂質(zhì)體來清除。三種巨噬細(xì)胞的清除效果通過流式或者免疫熒光來評價。

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），見圖a

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

Clo ICV：intra-cerebral ventricle injected clodronate liposomes (Clo ICV) ，腦室內(nèi)注射clodronate liposomes

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

見圖C：CD45和F4/80雙標(biāo)的流式圖，外周血循環(huán)單核細(xì)胞以10ul/g劑量通過腹腔注射來清除，相對于對照的6.7%，荷蘭Liposoma的巨噬細(xì)胞清除劑Clodronate liposomes氯膦酸鹽脂質(zhì)體可以清除94%以上的外周血單核細(xì)胞巨噬細(xì)胞。見圖d：小膠質(zhì)細(xì)胞通過長期飲食喂養(yǎng)PLX5622來清除；見圖e：腦駐留血管周圍巨噬細(xì)胞通過腦立體定位注射10ul荷蘭Liposoma的巨噬細(xì)胞清除劑Clodronate liposomes氯膦酸鹽脂質(zhì)體來清除。

論文信息：

論文題目：Macrophage lineage cells-derived migrasomes activate complement-dependent blood-brain barrier damage in cerebral amyloid angiopathy mouse model

期刊名稱：Nature Communications

時間期卷：14, Article number: 3945 (2023)

在線時間：2023年7月4日

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

產(chǎn)品信息：

貨號：CP-005-005

規(guī)格：5ml+5ml

品牌：Liposoma

產(chǎn)地：荷蘭

名稱：Clodronate Liposomes and Control Liposomes

辦事處：Target Technology(靶點(diǎn)科技)