摘要
目的 探索冠心康对氧化型低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)诱导的骨髓源巨噬细胞的小分子类泛素修饰物特异性蛋白酶1(small ubiquitin-related modifiers/sentrispecific proteases 1,SENP1)/信号传导蛋白和转录激活物(signal transducers and activators of transcription 3,STAT3)分子表达及胞葬作用的影响。方法 提取小鼠骨髓来源的单核细胞,加巨噬细胞集落刺激因子,诱导成巨噬细胞,显微镜下观察F4/80荧光标记;加入ox-LDL诱导,不同剂量的冠心康(1.25μg/m L、2.5μg/m L、5μg/m L)作用后,中性红试剂盒检测细胞吞噬率,实时荧光定量PCR(quantitative Real-time PCR,q RT-PCR)、免疫印迹法(Western blotting)检测胞葬相关分子TAM酪氨酸激酶受体(TYRO3/AXL/MER receptor tyrosine kinase)、血小板反应蛋白-1 (thrombospondin 1,THBS1)、乳脂球表皮生长因子8 (milk fat globule-epidermal growth factor,MFGE8)表达及SENP1/STAT3分子蛋白表达。结果 (1)骨髓源巨噬细胞的纯度约为99%;(2)与对照组比较,ox-LDL诱导的骨髓源巨噬细胞组细胞吞噬率显著降低(P <0.05);与模型组比较,冠心康显著上调细胞吞噬率(P <0.05);(3)与对照组比较,ox-LDL诱导的骨髓源巨噬细胞组胞葬相关分子、SENP1、p-STAT3表达降低(P <0.05);与模型组比较,冠心康显著上调这些分子的表达(P <0.05)。结论 冠心康通过上调SENP1/STAT3分子表达,增强骨髓源巨噬细胞的胞葬作用。
Objective Explore the effect of Guanxinkang on the expression of small ubiquitin-related modifiers/sentri-specific proteases 1(SENP1)/signal transducers and activators of transcription 3(STAT3) molecules and efferocytosis in bone marrow-derived macrophages induced by oxidized low density lipoprotein(ox-LDL).Methods Mononuclear cells derived from mice bone marrow were extracted,added with macrophage colony stimulating factor(M-CSF),and induced into macrophages.Bone marrow-derived macrophages were observed under microscope with F4/80 fluorescent labeling,and were induced by oxLDL and different doses of Guanxinkang(1.25 μg/mL,2.5 μg/mL,5 μg/mL).The cell phagocytosis rate was detected by the neutral red kit.The expressions of efferocytosis-related molecules,TYR03/AXL/MER receptor tyrosine kinase,Thrombospondin 1(THBS1),milk fat globule-epidermal growth factor(MFGE8) expression and SENP1/STAT3 molecular protein,were detected by Quantitative Real-time PCR(qRT-PCR) and Western blotting.Results(1) The purity of bone marrow-derived macrophages was about 99 %.(2) Compared with the control group,the phagocytic rate of bone marrow-derived macrophages induced by ox-LDL was significantly down-regulated(P <0.05).Compared with the model group,Guanxinkang significantly increased the phagocytic rate(P <0.05).(3) Compared with the control group,the expression of efferocytosis-related molecules,SENP1 and p-STAT3 decreased in bone marrowderived macrophages induced by ox-LDL(P <0.05).Compared with the model group,Guanxinkang significantly up-regulated the expressions of efferocytosis-related molecules,SENP1 and p-STAT3(P <0.05).Conclusion Guanxinkang enhanced the efferocytosis of bone marrow-derived macrophages by up-regulating the expression of SENP1/STAT3 signal pathway.
作者
张一凡
刘萍
ZHANG Yifan;LIU Ping(Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai,200032,China)
出处
《环球中医药》
CAS
2022年第10期1755-1760,共6页
Global Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81873117)。
