抑制S100A4表达在奥马珠单抗对IgE诱导肥大细胞活化中的作用  

Inhibition of S100A4 expression in omalizumab on IgE induced mast cell activation

作　　者：杨蕾[1] 李燕燕 樊长娥[2] 蔡慧娇 甄华[1] YANG Lei;LI Yan-yan;FAN Chang-e;CAI Hui-jiao;ZHEN Hua(Department of Pharmacy,Hohhot First Hospital,Hohhot 010030,Inner Mongolia Autonomous Region,China;Department of Allergy(Allergy)Response,Hohhot First Hospital,Hohhot 010030,Inner Mongolia Autonomous Region,China)

机构地区：[1]呼和浩特市第一医院药剂科,内蒙古自治区呼和浩特010030 [2]呼和浩特市第一医院变态(过敏)反应科,内蒙古自治区呼和浩特010030

出　　处：《中国临床药理学杂志》2024年第17期2513-2517,共5页The Chinese Journal of Clinical Pharmacology

基　　金：呼和浩特市第一医院院级科研基金资助项目(2022SYY053)。

摘　　要：目的探讨抑制S100A4表达增强奥马珠单抗(OmAb)对免疫球蛋白E(IgE)预结合肥大细胞的解离作用。方法将LAD2细胞随机分为正常组、IgE组(IgE诱导)、奥马珠单抗低剂量组(0.5 mg·mL^(-1)OmAb)、奥马珠单抗中剂量组(1.0 mg·mL^(-1)OmAb)、奥马珠单抗高剂量组(2.0 mg·mL^(-1)OmAb)、高剂量+si-S100A4组(转染si-S100A4+2.0 mg·mL^(-1)OmAb)。用流式细胞术检测细胞表面IgE水平,用β-氨基己糖苷酶释放测定法测量脱颗粒,用酶联免疫吸附试验(ELISA)法检测各组细胞组胺、白三烯C4水平,用蛋白质印迹法检测相关蛋白的表达水平。结果正常组、IgE组、奥马珠单抗高剂量组、高剂量+si-S100A4组处理6 h后细胞表面IgE水平分别为(4.13±0.52)%、(100.00±6.20)%、(60.12±3.41)%和(54.04±5.60)%,β-氨基己糖苷酶释放率水平分别为(12.59±1.35)%、(69.27±6.43)%、(45.39±2.14)%和(37.80±2.77)%,组胺水平分别为(2.43±0.16)、(8.57±0.41)、(4.91±0.24)和(3.01±0.23)ng·mL^(-1),白三烯C4水平分别为(198.85±18.91)、(423.56±1.25)、(273.68±17.11)和(242.79±12.44)pg·mL^(-1),磷酸化-细胞外信号调节激酶(p-ERK)蛋白相对表达水平分别为0.31±0.04、0.91±0.12、0.55±0.04和0.35±0.02;IgE组的上述指标分别与正常组比较,奥马珠单抗高剂量组的上述指标分别与IgE组比较,高剂量+si-S100A4组的上述指标分别与奥马珠单抗高剂量组比较,在统计学上差异均有统计学意义(均P<0.05)。结论抑制S100A4可增强OmAb对肥大细胞与IgE的解离作用,进一步阻断过敏介质的释放。Objective To investigate the effect of inhibiting S100A4 expression to enhance the dissociation of mast cells pre-bound by immunoglobulin E(IgE)by omalizumab(OmAb).Methods LAD2 cells were randomly divided into normal group,IgE group(IgE induction),OmAb-L group(0.5 mg·mL^(-1) OmAb),OmAb-M group(1.0 mg·mL^(-1) OmAb),OmAb-H group(2.0 mg·mL^(-1) OmAb),OmAb-h+si-S100A4 group(transfected with si-S100A4+2.0 mg·mL^(-1) OmAb).IgE levels on cell surface were detected by flow cytometry;degranulation was measured byβ-amino-hexosidase release assay;the levels of histamine and leukotriene C4 were detected by enzyme-linked immunosorbent assay(ELISA);the expression of related proteins was detected by Western blot.Results After 6 h treatment,IgE levels in normal group,IgE group,OmAb-H group and Om Ab-H+si-S100A4 group were(4.13±0.52)%,(100.00±6.20)%,(60.12±3.41)%and(54.04±5.60)%,respectively;β-amino-hexosidase release rates were(12.59±1.35),(69.27±6.43),(45.39±2.14)and(37.80±2.77)%,respectively;histamine levels were(2.43±0.16),(8.57±0.41),(4.91±0.24),(3.01±0.23)ng·mL^(-1),respectively;the C4 levels of leukotriene were(198.85±18.91),(423.56±1.25),(273.68±17.11)and(242.79±12.44)pg·mL^(-1),respectively;relative phosphorylated extracellular signal-regulated kinases(p-ERK)expression levels were 0.31±0.04,0.91±0.12,0.55±0.04 and 0.35±0.02,respectively.The above indexes in IgE group were compared with those in normal group,the above indexes of OmAb-H group were compared with IgE group,the above indexes of OmAb-H+si-S100A4 group were respectively compared with those of OmAb-H group,the differences were statistically significant(all P<0.05).Conclusion Inhibition of S100A4 can enhance the dissociation effect of OmAb on mast cells and IgE,and further block the release of allergic mediators.

关 键 词：奥马珠单抗 肥大细胞 S100钙结合蛋白A4 过敏反应 过敏介质 

分 类 号：R97[医药卫生—药品]