RBL-2H3细胞不适合用于类过敏模型的建立

目的考察RBL-2H3细胞是否适用于建立类过敏反应模型。方法用荧光定量聚合酶链反应考察RBL-2H3细胞上Mas相关G蛋白偶联受体(Mas-related G protein cou-pled receptor,Mrgpr)B2的表达情况;用显微镜观察和MTS法考察Compound 48/80对RBL-2H3细胞活力的影响;测定不同浓度Compound 48/80刺激RBL-2H3细胞脱颗粒释放的β-己糖胺酶含量,对比RBL-2H3细胞、人肥大细胞系LAD2和大鼠腹腔肥大细胞(rat peritoneal mast cells,RPMCs)对Compound 48/80响应性的差异。结果RBL-2H3细胞可表达MrgprB2受体。Compound 48/80能剂量依赖性地诱导RBL-2H3细胞释放β-己糖胺酶,但在高剂量(≥20 mg·L^-1)时对RBL-2H3的细胞活力产生明显影响,此时释放的β-己糖胺酶应当是由于其细胞毒作用引起细胞破裂所致。同在无毒剂量(10 mg·L^-1)的Compound 48/80刺激下,LAD2和RPMCs的响应性良好,β-己糖胺酶释放量分别为空白对照的15.02倍和16.05倍,而RBL-2H3细胞仅为空白对照的2.35倍。结论RBL-2H3细胞对Compound 48/80的响应性差,表明其并不适用于建立类过敏反应模型。

RBL-2H3 cells not suitable for pseudo-allergic reaction model

Aim To determine whether RBL-2H3 cells are suit-able for pseudo-allergic reaction model in vitro.Methods The expression of MrgprB2 receptor on RBL-2H3 cells was assayed by real-time fluorescence quantitative polymerase chain reaction.The effects of Compound 48/80 on the viability of RBL-2H3 cells were detected by the MTS assay and microscope.Superna-tantβ-hexosaminidase was assayed by using a fluorescent sub-strate 4-methylumbelliferyl N-acetyl-β-D-glucosaminide.Re-sults RBL-2H3 cells could express MrgprB2 receptor.Com-pound 48/80 could induce the release ofβ-hexosaminidase from RBL-2H3 cells in a concentration-dependent manner.However,the cell viability of RBL-2H3 was affected at high doses(≥20 mg·L^-1).In this context,the release ofβ-hexosaminidase might be attributed to its cytotoxicity.LAD2 cells and RPMCs showed high sensitivity in response to Compound 48/80 at a non-toxic concentration(10 mg·L^-1)with 15.02-fold and 16.05-fold differences,respectively.However,RBL-2H3 cells only re-leased 2.35-foldβ-hexosaminidase compared with control.Con-clusions In contrast to LAD2 cells and RPMCs,RBL-2H3 cells have a poor response to Compound 48/80,indicating that RBL-2H3 are not suitable for the pseudo-allergic reaction model in vitro.