β_(3)肾上腺素能受体激动剂对小鼠巨噬细胞胞葬的影响  

Effect of β_(3) adrenergic receptor agonist on efferocytosis in mouse macrophages

作　　者：刘欣妍 王越 王悦[1] 孙帅锋 吴小凡[1] LIU Xinyan;WANG Yue;WANG Yue;SUN Shuaifeng;WU Xiaofan(Department of Cardiology,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院心内科,北京100029

出　　处：《空军航空医学》2022年第6期307-310,共4页AVIATION MEDICINE OF AIR FORCE

基　　金：国家自然科学基金(82271605,82071573,81670317);北京市教育委员会科技计划重点项目(KZ202010025045)。

摘　　要：目的探讨β3肾上腺素能受体(β_(3) adrenoceptor,β_(3) AR)激动剂对小鼠巨噬细胞胞葬的影响及其可能机制。方法①紫外线照射诱导Jurkat细胞凋亡。紫外线分别照射Jurkat细胞60、75、90 min后,以Annexin V-FITC/PI标记,运用流式细胞仪检测Jurkat细胞凋亡率。②构建巨噬细胞胞葬模型。以荧光探针分别标记RAW264.7细胞和Jurkat细胞,将紫外线照射后凋亡Jurkat细胞加入巨噬细胞中共培养,构建巨噬细胞胞葬模型。运用流式细胞术分别比较加药组和对照组巨噬细胞胞葬率。③检测β_(3) AR及胞葬相关分子mRNA表达水平。运用实时荧光定量聚合酶链反应(RT-PCR)进一步检测对照组和1μmol/Lβ_(3) AR激动剂组β_(3) AR及胞葬相关分子mRNA表达水平。结果①与对照组比较,紫外线照射60、75、90 min后Jurkat细胞晚期凋亡率均显著升高[(8.17±1.27)%vs(39.7±14.63)%、(75.37±13.22)%、(79.23±11.99)%,P<0.05]。其中75 min组与90 min组较60 min组细胞凋亡率显著升高(P均<0.05),但75 min组和90 min比较差异无统计学意义(P=0.69)。②与对照组比较,0.1μmol/L和1μmol/Lβ_(3) AR激动剂组胞葬率均明显升高[(11.96±0.50)%vs(15.43±0.29)%vs(22.07±0.84)%,P<0.05],且以浓度依赖性方式增加。③β_(3) AR激动剂组β_(3) AR、MER酪氨酸蛋白激酶受体(Mer tyrosine kinase,Mertk)、生长停滞特异因子6 mRNA表达水平均明显高于对照组[(2.59±0.24)vs(1.00±0.21),P<0.05;(1.27±0.19)vs(1.00±0.04),P<0.05;(2.86±1.14)vs(1.00±0.12),P<0.05],但乳脂球表皮生长因子8 mRNA表达水平在2组间比较差异无统计学意义[(1.08±0.19)vs(1.00±0.21),P=0.55)]。结论β_(3) AR激动剂促进了小鼠巨噬细胞胞葬,其具体机制可能是通过上调Gas6、MerTK表达实现的。Objective To investigate the effect and possible mechanisms of β_(3) adrenoceptor(β_(3) AR)agonist on efferocytosis of mouse macrophages.Methods①Apoptosis of Jurkat cells induced by ultraviolet irradiation:Jurkat cells were divided into 60-,75-,and 90-minute groups after ultraviolet irradiation,labeled with Annexin V-FITC/PI.Flow cytometry was used to detect the apoptosis rate of Jurkat cells.②Macrophage efferocytosis model:RAW264.7 cells and Jurkat cells were labeled with fluorescent probes.Apoptotic Jurkat cells induced by ultraviolet were co-cultured with macrophages to construct an efferocytosis model.Flow cytometry was used to compare the efferocytosis rate of the β_(3) AR agonist groups and control group.③mRNA levels of β_(3) AR and efferocytosis-related molecules:Real-time fluorescence quantitative polymerase chain reaction(RT-PCR)was used to detect the mRNA levels of β_(3) AR and efferocytosis-related molecules in the control group and 1umol/L β_(3) AR agonist group.Results①Compared with the control group,the apoptosis rate of Jurkat cells was significantly increased after 60,75 and 90 minutes of ultraviolet irradiation[(8.17±1.27)%vs(39.7±14.63)%,(75.37±13.22)%,(79.23±11.99)%,all P<0.05].The apoptosis rate of the 75-minute group and 90-minute group was significantly higher than that of the 60-minute group(P<0.05).However,there was no significant difference between the 75-minute group and the 90-minute group(P=0.69).②Compared with the control group,the efferocytosis rate of 0.1μmol/L and 1μmol/L β_(3) AR agonist groups was significantly increased[(11.96±0.50)%vs(15.43±0.29)%and(22.07±0.84)%,both P<0.05],and improved in a concentration-dependent manner.③mRNA expression levels of β_(3) AR,Mer TK and Gas6 in the β_(3) AR agonist group were significantly higher than those of the control group[(2.59±0.24)vs(1.00±0.21),P<0.05,(1.27±0.19)vs(1.00±0.04),P<0.05,(2.86±1.14)vs(1.00±0.12),P<0.05].However,there was no significant difference in expression levels of MFG-E8 mRNA between

关 键 词：β_(3)肾上腺素能受体 胞葬 巨噬细胞 

分 类 号：R541.4[医药卫生—心血管疾病]