钙敏感受体对人单核巨噬细胞株THP-1细胞因子分泌功能的影响

目的 探讨钙敏感受体(Calcium sensing receptor,CaSR)对THP-1源性巨噬细胞分泌细胞因子能力的影响。方法 首先将人单核细胞株THP-1用佛波脂(PMA,100 nmol/L)诱导,使之转化为具有贴壁能力的巨噬细胞株THP-1(以下简称THP-1)。用CaSR特异激动剂氯化钆(GdCl₃,1 mmol/L)处理细胞后,用ELISA法检测CaSR的激活对THP-1分泌细胞因子IL-1β、TNFα的影响,并利用Western Blotting方法检测NF-κB信号通路中p65、p50和IκBα蛋白的表达变化。结果 (1)用GdCl₃处理后,THP-1细胞上清液中IL-1β、TNF-α的含量明显升高,但该作用可被CaSR特异抑制剂NPS2390(10 μmol/L)抑制;(2)在Gd³⁺作用组,THP-1细胞中NF-κB-p65表达明显上调,同时伴有IκBα的减少;在NPS2390预处理组,Gd³⁺所引起的NF-κB-p65和IκBα变化被抑制;各组NF-κB-p50变化不明显。结论 CaSR的激活可能通过激活NF-κB信号通路促进THP-1源性巨噬细胞株分泌细胞因子IL-1β、TNF-α,提示CaSR可能参与涉及单核巨噬细胞激活的各种疾病。

The effect of calcium-sensing receptor on the secretion of cytokine in the differentiated THP-1 cells

Objective To observe the function of calcium sensing receptor（CaSR） in differentiated THP - 1 （human acute monocytic leukemia cell line）cells. Methods In the present study,we induced and differentiated the monocyte - like THP - 1 ceils into maerophage - like THP - 1 cells with PMA. The secretion of cytokines was analyzed by ELISA. The expression of CaSR and NF - κB in the differentiated THP - 1 cells was identified by Western blotting. Results （ 1 ） We found that GdCI3 elevation stimulated both of IL - 113 and TNFet release in differentiated THP- 1 ceils. （2）GdC13 could increase the expression of NF- κB -p65 and decrease the expres- sion of NF - κB - IκBα. The effects of GdC13 were inhibited by NPS2390. Conclusion The activation of CaSR in macrophage -like THP - 1 ceils could commit to cytokine secretion via NF -κB signal transduction pathway. These results suggested that CaSR might be involved in a variety of pathological processes mediated by activated monocyte - macrophages,