降钙素基因相关肽通过抑制Nod样受体蛋白3表达促进小鼠成骨细胞分化的研究

目的 研究降钙素基因相关肽（CGRP）作用下相关炎症体和信号因子的表达变化，探讨CGRP对成骨细胞分化的作用机制。方法 将不同浓度的CGRP（0、10、30、100 ng•mL-1）加入到成骨细胞中，采用实时聚合酶链反应技术检测细胞内Nod样受体蛋白3（NLRP3）及白细胞介素-1β（IL-1β）mRNA的表达水平，蛋白质印迹法检测NLRP3的蛋白表达，酶联免疫吸附测定检测IL-1β的蛋白表达，流式细胞仪检测细胞内活性氧（ROS）含量，茜素红染色显示成骨细胞分化情况。结果 随着CGRP浓度的增加，NLRP3和IL-1β的蛋白表达及mRNA表达均呈降低趋势（P<0.05），而且细胞内ROS浓度逐渐下降（P<0.05）。100 ng•mL-1CGRP实验组较0 ng•mL-1CGRP对照组显著促进成骨细胞分化。结论 CGRP在一定条件下，可通过抑制细胞内炎症因子的表达促进成骨细胞分化。

Calcitonin gene-related peptide inhibits the expression of Nod-like receptor protein 3 to promote osteoblast differen-tiation in mouse osteoblasts in vitro

Objective This study aims to investigate the regulatory effects of calcitonin gene-related peptide (CGRP) on Nod-like receptor protein 3 (NLRP3) and interleukin-1β (IL-1β) to promote osteoblast differentiation. Methods Different concentrations of CGRP (0, 10, 30, 100 ng·mL-1) were added to mouse osteoblasts in vitro. The mRNA and protein expression levels of both NLRP3 and IL-1β were examined using Real-time polymerase chain reaction and Western blot, respectively. Moreover, the concentrations of IL-1β protein and intracellular reactive oxygen species (ROS) were detected using enzyme-linked immunosorbent assay and flow cytometry, respectively. The osteogenic differentiation of mouse osteoblasts was identified through alizarin red staining. Results The protein and mRNA expression levels of both NLRP3 and IL-1β significantly decreased (P<0.05) with increasing CGRP concentration. Moreover, the contents of intracellular ROS gradually decreased (P<0.05). The osteogenic differentiation of the osteoblasts was more enhanced in the group treated with 100 ng·mL-1 CGRP than in the empty group (0 ng·mL-1CGRP). Conclusion CGRP promotes osteoblast differentiation by inhibiting the expression of inflammatory factors.