甘露聚糖誘導的銀屑病
攜帶Ncf1基因突變的小鼠經鼻內給予20 mg甘露聚糖,以誘導銀屑病和銀屑病關節炎(PsA)(24)。采用與前述相同的評分系統,每日對關節炎嚴重程度進行評分。銀屑病樣皮膚表現的嚴重程度通過一套15分制系統進行監測,其中每只耳朵或每個爪子按1至3分的等級進行評估(1分,輕微脫皮;2分,中度脫皮;3分,嚴重脫皮并伴有部分脫毛)(24)。
對于體內巨噬細胞清除實驗,在誘導關節炎前2天,經鼻內給予小鼠50 μl氯膦酸二鈉脂質體或對照PBS脂質體(荷蘭Liposoma BV公司)。通過流式細胞術分析評估清除效率。
對于體內Ym1蛋白補充實驗,在誘導關節炎的第0天經鼻內給予小鼠甘露聚糖,并在誘導后的第0、2和4天經鼻內給予重組Ym1蛋白(每只小鼠1 μg;北京義翹神州生物技術有限公司)。
小鼠肺部炎癥模型
對于甘露聚糖誘導的肺部炎癥模型,BR.Ncf1和BR.Ncf1.Ym1Δ小鼠經鼻內給予甘露聚糖(每只小鼠0.5 mg)。分別于第0、2和5天處死小鼠。計算肺重與體重的比值(肺指數)。制備肺組織進行蘇木精-伊紅(H&E)染色。通過流式細胞術分析支氣管肺泡灌洗液(BALF)中的炎癥細胞。
對于過敏性氣道肺部炎癥(AIPI)模型的誘導,B10.RIII和BR.Ym1Δ小鼠在第0天和第7天通過腹腔注射0.2 ml乳化液進行致敏,該乳化液由10 μg(溶于100 μl)卵清蛋白(OVA)(美國Sigma-Aldrich公司)和100 μl Imject Alum佐劑(美國Pierce,Thermo Fisher Scientific公司)混合而成。在第14天至第20天期間,小鼠每天接受1% OVA氣溶膠激發30分鐘。對照組小鼠接受假性致敏,并暴露于相同體積的PBS。在第21天處死小鼠。制備肺組織用于組織學分析和細胞因子表達檢測。通過流式細胞術分析BALF中的炎癥細胞。收集血清,用于檢測Ym1和OVA特異性IgG1及IgE的濃度。
肺泡巨噬細胞清除效果檢測

論文信息:
論文題目:Natural polymorphism of Ym1 regulates pneumonitis through alternative activation of macrophages
期刊名稱:Science Advances
時間期卷:Vol 6, Issue43(2020)
在線時間:2020年10月21日
DOI: 10.1126/sciadv.aba9337
產品信息:
貨號:CP-005-005
規格:5ml+5ml
品牌:Liposoma
產地:荷蘭
名稱:Clodronate Liposomes&Control Liposomes
辦事處:靶點科技
Clodronate Liposomes氯膦酸鹽脂質體清除肺炎模型小鼠體內肺泡巨噬細胞。荷蘭Liposoma巨噬細胞清除劑ClodronateLiposomes見刊于Science Advances:Ym1的自然多態性通過巨噬細胞的替代性激活來調節肺炎。

Liposoma巨噬細胞清除劑Clodronate Liposomes氯膦酸二鈉脂質體清除巨噬細胞的材料和方法:ClodronateLiposoems氯膦酸鹽脂質體清除甘露聚糖誘導的肺部炎癥模型巨噬細胞模型
In vivo macrophage depletion
Mice with Ncf1 mutation were administrated intranasally with 20 mg of mannan to induce psoriasis and PsA (24) and were scored daily for arthritis severity using the same scoring system as mentioned above. The severity of the psoriasis-like skin manifestations was monitored on a 15-score system, in which ears or each paw was evaluated by a scale ranging from 1 to 3 (1, weak skin peeling; 2, moderate skin peeling; and 3, heavy skin peeling with some hair loss) (24). For the in vivo macrophage depletion experiment, 50 μl of clodronate-liposome or control PBS-liposome (Liposoma BV, The Netherlands) was administrated intranasally to mice 2 days before arthritis induction. The depletion efficiency was assessed by flow cytometry analysis. For the in vivo Ym1 protein supplement experiment, mice were intranasally administrated with mannan to induce arthritis at day 0, and recombinant Ym1 protein (1 μg per mouse; Sino Biological Inc.) was treated intranasally at days 0, 2, and 4 after arthritis induction.
巨噬細胞清除材料和方法文獻截圖:ClodronateLiposoems氯膦酸鹽脂質體清除甘露聚糖誘導的肺部炎癥模型巨噬細胞模型