目的探讨利拉鲁肽对高糖诱导的单核细胞亚群分化的影响及其机制。方法制备小鼠脾脏原代细胞悬液,高糖刺激诱导单核细胞亚群分化,进一步采用利拉鲁肽进行干预,利用流式细胞仪检测单核细胞亚群分化情况,依据Ly6C的表达量将单核细胞分为Ly6Clow,Ly6Cmid,Ly6Chi3个亚群。再以2’,7’-二氯荧光黄-二醋酸盐(DCFH-DA)标记细胞,通过流式细胞仪检测细胞内二氯荧光黄(DCF)的荧光强度,评估各单核细胞亚群的活性氧簇(ROS)水平。结果15,25,35mmol/L葡萄糖可浓度依赖性地促进Ly6Chi和Ly6Cmid单核细胞亚群分化,且能增加Ly6C+单核细胞内ROS的产生,而对Ly6C-单核细胞中ROS的水平没有影响。100U、200U利拉鲁肽浓度依赖性地抑制高糖诱导的Ly6Cmid和Ly6Chi单核细胞亚群分化,并使Ly6Clow单核细胞亚群明显增加。利拉鲁肽显著抑制高糖诱导的Ly6C+单核细胞中ROS平,但对Ly6C-单核细胞中的ROS无影响。结论利拉鲁肽可抑制高糖诱导的炎症单核细胞亚群分化,并减少ROS的产生。
Objective To explore the mechanism by which liraglutide modulated the differentiation of monocyte subsets in high glucose (HG) conditions. Methods Primary mouse splenocyte suspensions were cultured in HG conditions induced by IFN-γ in the presence or absence of liraglutide. The cells were harvested, co-incubated with antibodies, and analyzed on a BD FACS Calibur. Mononuclear cells (MNCs) were gated according to lower granularity and larger size by SSC and FSC. Monocytes (MCs) were defined as CDllb+MNC and divided into three subsets based on Ly6C expression : Ly6Ck^low, Ly6C^mid, and Ly6C^hi. ROS production in Ly6C^+ and Ly6C^- MCs was detected by 2', 7' - (DCFH-DA) staining and ROS-containing MCs were identified as DCF cells in both Ly6C+ and Ly6C-MCs. Results HG ( 15 mmol/L,25 mmol/ L, 35 mmol/L) dose-dependently increased Ly6C^hi and Ly6C^mid MC differentiation and also enhanced the production of ROS in Ly6C+MCs. Liraglutide (100 U, 200 U) dose-dependently inhibited HG-induced Ly6C^hi and Ly6C^mid MC differentiation and also promoted the differentiation of Ly6C^kw MCs. Moreover, liraglutide significantly inhibited HG-induced ROS production in Ly6C^+ MCs. Conclusion Liraglutide treatment significandy inhibited inflammatory MC differentiation induced by HG and also reduced ROS production in inflammatory MCs.