过表达TRPV6基因对小鼠破骨细胞钙转运基因与凋亡相关基因表达的影响

Effect of overexpression of the transient receptor potential vanilloid 6 gene on apoptosis and calcium transport in mouse osteoclasts

破骨细胞是体内唯一的骨吸收细胞,对钙离子跨细胞膜转运及维持正常血钙水平有非常重要的作用。瞬时性受体电位通道香草酸受体6(transient receptor potential vanilloid receptor 6,TRPV6)是TRP家族中的一员,对钙离子有高度选择性,且是钙离子跨细胞转运的限速门控通道。本试验旨在研究TRPV6过表达对破骨细胞钙转运相关基因及细胞凋亡相关基因表达的影响。用TRPV6过表达慢病毒载体(LV-TRPV6)转染培养7 d的破骨细胞构建TRPV6过表达破骨细胞模型,以阐释小鼠破骨细胞中TRPV6与钙离子转运和细胞凋亡的关系。将破骨细胞分为3组:空白对照组、阴性对照组和LV-TRPV6组。结果显示:与空白对照组比较,破骨细胞TRPV6过表达后钙离子转运相关基因(calbindin-D28K,NCX1) mRNA和蛋白表达水平显著增加(P<0.05);破骨细胞TRPV6过表达后细胞凋亡相关基因mRNA和蛋白表达水平显著降低(Fas和FasL除外)(P<0.05)。流式细胞仪分析显示,LV-TRPV6组凋亡率显著下降6.33%±0.235%(P<0.05)。研究表明,破骨细胞过表达TRPV6基因导致钙转运相关基因表达增加并抑制破骨细胞凋亡。

Osteoclasts are the only bone-absorptive cells in the body and are vital in calcium transmembrane transport and maintenance of normal serum calcium levels.TRPV6,a member of the TRP family,is a highly selective Ca2+channel and plays an important role in calcium transportation.This study was conducted to investigate the effect of TRPV6 overexpression on apoptosis and calcium transport in mouse osteoclasts.After osteoclasts cultured for 7 d,a TRPV6 overexpression lentivirus vector(LV-TRPV6) was used to transfect osteoclasts and establish a TRPV6 osteoclast overexpression model to explore the relationship between TRPV6 and calcium transport and apoptosis in mouse osteoclasts.The osteoclastic cells were divided into three groups:the blank control,the negative control and the LV-TRPV6 group.The rresults showed that compared with the control group,the relative mRNA and protein expression of calcium transport-related genes(calbindin-D28 K,NCX1) increased significantly(P<0.05) following TRPV6 overexpression.Additionally,the relative mRNA and protein expression of apoptosis-related genes decreased significantly(P<0.05),except Fas and Fas L.The flow cytometry analysis showed that the apoptotic rate of the LV-TRPV6 group decreased to 6.33%±0.235%(P<0.05).This study indicated that TRPV6 overexpression enhanced certain calcium transport-related genes expression and inhibited apoptosis in mouse osteoclasts.