Physiological role of CD206 macrophages and NAD+ metabolism in obesity and insulin resistance.

首先，我成功地创造了基因工程CD206+ m2样巨噬细胞特异性Nampt敲除（KO）转基因小鼠。我发现他莫昔芬（TAM）的施用不影响小鼠的体重。服用TAM后，我进行了ipGTT（腹腔葡萄糖耐量试验）和ITT（胰岛素耐量试验）。我的数据显示，CD206特异性Nampt-KO小鼠与正常食饲条件下的对照小鼠之间没有显著差异。然后给这些小鼠喂食高脂肪饮食（HFD）。与我的假设一致，Nampt-KO小鼠比对照小鼠体重增加更多。这些数据强烈提示CD206+巨噬细胞通过nampt衍生的NAD+调节饮食诱导的肥胖或体重增加，并伴有糖代谢恶化。我的数据表明，CD206基因在肥胖状态下对脂肪组织代谢的影响更差，Nampt KO进一步加重了这种影响。然而，潜在的分子机制尚不清楚。我的数据还显示，cd206阳性的巨噬细胞在喂食hfd 12周后显示炎症基因标记（Il1b, Tnfa, CD11c）。接下来，我将在CD206-/-（CD206基因敲除）小鼠上进行研究，以评估CD206 M2巨噬细胞如何参与肥胖条件下脂肪组织代谢的调节。我将对这些肥胖小鼠进行GTT和ITT，并在牺牲小鼠后收获组织。我将对FACS分选的CD206+巨噬细胞进行免疫组织化学、RNA提取、western blotting、流式细胞术、qPCR等方法来阐明其潜在的机制。

First, I have successfully create of genetically engineered CD206+ M2like-macrophage-specific Nampt knock-out (KO) transgenic mice. I found that tamoxifen (TAM) administration did not affect the body weight of the chow-fed mice. After the administration of TAM I have performed ipGTT (intraperitoneal glucose tolerance test) and ITT (insulin tolerance test). My data showed that there is no significant difference between CD206 specific Nampt-KO mice and control mice normal chow-fed conditions. Then these mice were put on high fat diet (HFD). In consistent with my hypothesis, Nampt-KO mice gaining more body weight compared to the control mice. This data strongly suggested that CD206+ macrophages regulated diet-induced obesity or weight gain via Nampt-derived NAD+, with worsen glucose metabolism. My data suggest that CD206 gene have worsen effect on adipose tissue metabolism in obese states, which further worsen by Nampt KO. However, underlying molecular mechanism is unknown. My data also showed that CD206-positive macrophages shows inflammatory gene markers (Il1b, Tnfa, CD11c) after 12 weeks of HFD-feeding. Next, I am going to work on CD206-/-(CD206 gene knock-out) mice to evaluate how CD206 M2 macrophages is involved in regulation adipose tissue metabolism in obese conditions. I will perform GTT and ITT of these obese mice, and will harvest the tissue after sacrifice the mice. I will perform immunohistochemistry, RNA extraction, western blotting, flow cytometry, the qPCR of FACS sorted CD206+ macrophages to elucidate the underlying mechanism.