经方丹栀逍遥丸对代谢相关脂肪性肝病小鼠巨噬细胞抗慢炎状态的调节作用

Regulation of Classical Prescription “Danzhi Xiaoyao Pill(丹栀逍遥丸)”on Anti-Chronic Inflammatory State of Macrophages in Mice with Metabolic Associated Fatty Liver Disease

目的 评价经方丹栀逍遥丸(JF-DZXYW)对代谢相关脂肪性肝病小鼠巨噬细胞抗慢性炎症的调控作用,对巨噬细胞吞噬能力、巨噬细胞表面标志物MHC-Ⅱ、CD86、CD80、TLR4表达及巨噬细胞内活性氧表达(ROS)的影响,并阐述其诱导免疫系统发挥抗炎症作用。方法 40只体质量18~22 g雄性昆明种小鼠,随机分为对照组(生理盐水组)、JF-DZXYW低剂量组(125μg/mL)、JF-DZXYW中剂量组(250μg/mL)和JF-DZXYW高剂量组(500μg/mL),灌胃0.4 mL/t/d,共15 d。第16天小鼠行腹腔注射0.2 mL 10%蛋白胨,3 d后小鼠腹腔注射新配制的5%鸡红细胞各0.5 mL,提取小鼠腹腔巨噬细胞进行巨噬细胞吞噬功能检测、细胞表面标志物流式细胞分析及ROS表达检测。结果 3个剂量组的JF-DZXYW汤剂吞噬百分率及吞噬指数均显著高于对照组(P<0.01)。JF-DZXYW低、中、高剂量组小鼠腹腔巨噬细胞表面标志物MHC-Ⅱ、CD86、CD80表达明显高于对照组(P<0.01),JF-DZXYW中、高剂量组小鼠腹腔巨噬细胞表面标志物TLR4表达均明显高于对照组(P<0.01)。JF-DZXYW低剂量组小鼠腹腔巨噬细胞内ROS表达明显高于对照组(P<0.05),JF-DZXYW中、高剂量组小鼠腹腔巨噬细胞内ROS表达均明显高于对照组(P<0.01)。结论 JF-DZXYW可以通过上调巨噬细胞功能、促巨噬细胞表面标志物MHC-Ⅱ、CD86、CD80、TLR4及巨噬细胞内活性氧表达提升机体免疫功能从而抑制慢性炎症状态。

Objective To evaluate the regulatory effect of the classical prescription“Danzhi Xiaoyao Pill(丹栀逍遥丸,JF-DZXYW)”on the anti-chronic inflammation of macrophages in mouse with metabolic associated fatty liver disease,its effect on the phagocytic ability of macrophages,the expressions of macrophage surface markers MHC-II,CD86,CD80 and Toll-like receptor 4(TLR4)and the expression of reactive oxygen species(ROS)in macrophages,and to elaborate its induction of the immune system to exert anti-inflammatory effects.Methods Forty male Kunming mice(18~22 g)were randomly divided into a control group(saline group),JF-DZXYW low dose group(125μg/mL),JF-DZXYW medium dose group(250μg/mL)and JF-DZXYW high dose group(500μg/mL)with 0.4 for 15 d.On the 16th day,the mice were intraperitoneally injected with 0.2 mL of 10%peptone.Three days later,the mice were intraperitoneally injected with 0.5 mL of newly prepared 5%chicken red blood cells,and peritoneal macrophages were extracted for macrophage phagocytic function testing,cell surface marker flow cytometry analysis and ROS expression detection.Results The phagocytosis percentage and phagocytosis index of JF-DZXYW three dose groups were significantly higher than those in the control group(P<0.01).The expressions of the surface markers MHC-II,CD86 and CD80 in the peritoneal macrophages of mice in the JF-DZXYW low,middle and high dose groups were significantly higher than those in the control group(P<0.01).The expression of the surface markers TLR4 in the peritoneal macrophages of mice in the JF-DZXYW middle and high dose groups were significantly higher than those in the control group(P<0.01).The expression of TLR4,a surface marker,was significantly higher in mice in the JF-DZXYW low-dose group than that in the control group(P<0.05),and the expression of ROS in mice in the JF-DZXYW middle and high dose groups was significantly higher than that in the control group(P<0.01).Conclusion JF-DZXYW can enhance the immune function of the body through up-regulating macrophage function,promoting macrophage surface markers MHC-II,CD86,CD80 and TLR4 and intra-macrophage ROS expression,thus suppressing the chronic inflammatory state.