摘要
目的观察β-肾上腺素受体(AR)激动药对人成纤维样滑膜细胞(FLS)增殖的作用,探讨β-AR信号对人FLS功能的影响。方法组织块培养法培养人FLS;使用肿瘤坏死因子α(TNF-α)作为刺激剂,MTT法分别检测β1-AR选择性激动剂多巴酚丁胺和β2-AR选择性激动剂沙丁胺醇对人FLS增殖作用的影响,并计算增殖抑制率;使用沙丁胺醇+β2-AR选择性拮抗剂ICI 118551观察其对FLS增殖功能的影响;Western blot法检测人FLSβ-AR、G蛋白偶联受体激酶2(GRK2)和β抑制蛋白2(β-arrestin2)胞膜蛋白的表达水平。结果 TNF-α可以显著促进人FLS的增殖;β2-AR激动剂沙丁胺醇能明显抑制FLS的增殖,其拮抗剂ICI 118551能显著拮抗沙丁胺醇的作用;β1-AR选择性激动剂多巴酚丁胺对FLS的增殖无显著影响;TNF-α和沙丁胺醇均能显著降低β2-AR胞膜表达,上调GRK2、β-arrestin2的胞膜表达。结论β2-AR信号的激活可以抑制FLS增殖;TNF-α和沙丁胺醇可以使GRK2、β-arrestin2的胞膜表达增加,β2-AR胸膜表达下降。
Objective To observe the function of β-adrenergic receptor （ AR） agonists on human fibroblasts syno-viocytes （ FLS） proliferation and explore the effect ofβ-AR signaling on FLS function. Methods Tissue culturing method cultured human FLS;using TNF-α as a stimulant;MTT method was used to detect the effect ofβ1-AR se-lective agonist dobutamine andβ2-AR selective agonist salbutamol on the proliferation of human FLS, and calculate the proliferation inhibition rate. β2-AR selective antagonist ICI 118551 was used to observe the effect on FLS prolif-eration; Western blot method could detect cell membrane protein expression level belonged to FLS β-AR,GRK2 and β-arrestin2 on human. Results TNF-αcould significantly promote the proliferation of FLS on human,β2-AR agonist salbutamol could significantly inhibit the proliferation of FLS, and its antagonist ICI 118551 could signifi-cantly inhibit salbutamol action. β1-AR selective agonist dobutamine had no significant effect on the proliferation of FLS;TNF-α and salbutamol significantly decreased β2-AR expression of cell membrane, and up-regulated GRK2 andβ-arrestin2 expression in cell membrane. Conclusion The activation ofβ2-AR signal can inhibit the prolifera-tion of FLS;TNF-α and salbutamol can increase the expression of GRK2,β-arrestin2,and decrease the expression of β2-AR in cell membrane.
出处
《安徽医科大学学报》
CAS
北大核心
2015年第7期978-981,共4页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81173075,81330081,81302784)
安徽省科技攻关计划项目(编号:1301042098)
关键词
肾上腺素受体
成纤维样滑膜细胞
沙丁胺醇
增殖
adrenergic receptor
fibroblast-like synoviocytes
albuterol sulfate
desensitization
