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本研究探讨Gpr97在哮喘细胞模型中对细胞因子IL-4、IL-6分泌的调控作用。分离野生型和Gpr97-/-小鼠的骨髓细胞后,通过细胞因子诱导其分化为肥大细胞,光镜下观察细胞形态,FACS法检测肥大细胞的纯度。利用含IgE的血清致敏、卵清蛋白(OVA)激活肥大细胞,通过PCR和ELISA检测Gpr97对肥大细胞分泌IL-4、IL-6的影响。结果发现:肥大细胞表面标志物APC-CD117,FITC-FcεRIα的双阳性率均达到了67%以上。OVA作用后野生型(WT)小鼠肥大细胞Gpr97表达显著上调(P<0.001),Gpr97-/-鼠肥大细胞分泌IL-4和IL-6的量在mRNA和蛋白水平都显著低于野生型小鼠。以上结果提示Gpr97能够通过增加肥大细胞分泌IL-4和IL-6从而加重哮喘的发生。
This study was to investigate the regulatory effect of Gpr97 on the secretion of IL-4 and IL-6 in asthmatic models. The bone marrow cells of wild-type and Gpr97 - / - mice were isolated and induced to differentiate into mast cells by cytokines. The morphology of the cells was observed under light microscope. The purity of mast cells was detected by FACS. Mast cells were activated with ovalbumin (OVA) by using serum containing IgE, and the effect of Gpr97 on the secretion of IL-4 and IL-6 by mast cells was detected by PCR and ELISA. The results showed that the double positive rate of mast cell surface markers APC-CD117 and FITC-FcsRIa reached more than 67%. The expression of Gpr97 in mast cells of wild-type (WT) mice was significantly upregulated after OVA treatment (P <0.001), and the levels of IL-4 and IL-6 secreted by Gpr97 - / - mouse mast cells at mRNA and protein levels were significantly lower than those of wild type Mouse. The above results suggest that Gpr97 can exacerbate asthma by increasing the secretion of IL-4 and IL-6 by mast cells.