鸟结核分枝杆菌刺激巨噬细胞后对细胞骨架蛋白β-actin的调控机制研究

Regulatory mechanisms of the cytoskeletal protein β-actin after stimulation of Mycobacterium aviumto macrophages

目的研究鸟结核分枝杆菌刺激巨噬细胞后其对巨噬细胞骨架蛋白及其调节蛋白的作用机制。方法 RT-PCR方法分析M.avium刺激巨噬细胞后cofilin-1,β-actin基因的表达水平,同时Western blot方法从蛋白水平分析M.avium刺激巨噬细胞后β-actin、cofilin-1蛋白的表达水平。流式细胞术检测M.avium刺激巨噬细胞后巨噬细胞凋亡及坏死的情况。结果β-actin基因及其蛋白在巨噬细胞受M.avium刺激后表达显著下调,而β-actin蛋白的调节蛋白cofilin-1表达显著增强。同时,M.avium刺激巨噬细胞后诱导巨噬细胞凋亡显著增多。结论 M.avium刺激巨噬细胞后可通过调控β-actin及cofilin-1蛋白的表达,从而诱导巨噬细胞凋亡或坏死。

We investigated the functions of the cytoskeleton proteins and their regulatory proteins in Mycobacterium ctvium （M. avium） stimulation macrophages. The expression of β- actin and cofilin-1 were detected in mRNA and proteins levels by RT-PCR and Western blot technologies, respectively. Meanwhile, the apoptosis or necrosis of macrophages stimulated with M. aviura was analyzed by flow cytometry. Results showed that β-actin mRNA and proteins o{ ~actin were both down-regula- ted in M. avium stimulation macrophages, however the expression ofcofilin 1 gene and proteisn was presented oppositely both in mRNA and protein level compared with the expression of β-actin. Furthermore, the apoptosis or necrosis of macrophages in- duced by M. avium was increased significantly. We confirmed that the expression of β-actin and confilin-1 could be regulated by M. avium stimulation to induce theapoptosis or necrosis of macrophages.