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  • Title: [Bortezomib inhibits hypoxia-induced increase of Orai-1 expression in pulmonary arterial smooth muscle cell].
    Author: Xu L, Tian GY, Wang LH, Liu YB, Gao ZF, Li GH, Fu XH.
    Journal: Zhonghua Jie He He Hu Xi Za Zhi; 2017 Feb 12; 40(2):118-122. PubMed ID: 28209043.
    Abstract:
    Objective: In this study, a primary culture system for the rat distal pulmonary arterial smooth muscle cell (PASMC) was established to observe the effect of Bortezomib a treatment on the basal intracellular calcium concentration ([Ca(2+) ](i)), store operated calcium entry (SOCE) and Orai-1 expression in rat PASMC. Methods: We employed the primary culture method for the rat distal PASMC including the enzymatically dissociation of PASMC from the freshly isolated distal pulmonary artery and the culture of PASMC. The In Cyte system was used to measure the basal [Ca(2+) ](i) and SOCE after substantial treatment.Orai-1 protein expression in rat pulmonary artery smooth muscle was detected by Western blot. Results: Compared with Hypoxia group, the basal [Ca(2+) ](i) were significantly reduced in Hypoxia+ BTZ group(P<0.01). The basal [Ca(2+) ](i) A340/A380 ratio of Normoxia group was(1.07±0.02). The basal [Ca(2+) ](i) of Hypoxia group was(1.49±0.03); The Hypoxia+ BTZ group was(1.17±0.03). Compared with Hypoxia group, the store operated calcium entry were significantly reduced in Hypoxia+ BTZ group(P<0.01). The SOCE A340/A380 ratio of Normoxia group was(0.56±0.02). The SOCE of Hypoxia group was(0.84±0.02); The Hypoxia+ BTZ group was(0.66±0.02). The level of Orail-1 protein in pulmonary artery smooth muscle of Hypoxia group was (181.5±12.7)% higher than control group which was(100±0)%, (P<0.05). In the Hypoxia+ BTZ group Orai-1 protein expression was recovered(146.7±15.1)%, (P<0.05). Conclusion: Bortezomib inhibit chronically hypoxic enhancement of Orail-1 protein expression, basal [Ca(2+) ](i) and SOCE in rat distal pulmonary arterial smooth muscle cells. 目的：探讨蛋白酶体抑制剂硼替佐米对低氧诱导的肺动脉平滑肌细胞中钙池操纵性钙内流(SOCE)通道的重要组成蛋白Orai－1表达的影响。 方法：原代培养肺动脉平滑肌细胞，进行细胞鉴定，将细胞分为常氧组、低氧组及低氧＋硼替佐米组，60 h后钙离子成像系统实时测定细胞基础钙及SOCE。通过免疫印迹法测定各组肺动脉平滑肌细胞Orai－1的表达情况。 结果：常氧组细胞基础钙A340/A380为(1.07±0.02)，低氧组细胞基础钙比值为(1.49±0.03)，低氧＋硼替佐米组细胞的基础钙比值为(1.17±0.03)；与低氧组比较，低氧＋硼替佐米组细胞基础钙明显降低(P<0.01)。常氧组细胞SOCE的A340/A380比值为(0.56±0.02)，低氧组细胞SOCE比值为(0.84±0.02)，而低氧＋硼替佐米组细胞的SOCE为(0.66±0.02)；与低氧组比较，低氧＋硼替佐米组细胞SOCE明显降低(P<0.01)。与常氧组相比低氧组、低氧＋硼替佐米组细胞Orai－1蛋白的相关表达量分别为(181.5±12.7)%和(146.7±15.1)%，低氧＋硼替佐米组细胞Orai－1蛋白表达量相较于低氧组明显降低(P<0.05)。 结论：硼替佐米可以明显抑制低氧引起的大鼠肺动脉细胞Orai－1表达的上调，降低基础钙及SOCE。.
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