【摘要】 目的观察近端肾小管上皮细胞对成骨细胞骨代谢相关基因表达的直接调节作用。方法体外分离培养大鼠近端肾小管上皮细胞与头盖骨成骨细胞;用甲状旁腺激素(PTH)、137Cs-γ射线及25(OH)D3预作用前者,再与后者分为4组共培养:①单培养组(成骨细胞单培养)、②共培养组(肾小管上皮细胞与成骨细胞共培养)、③刺激共培养组(肾小管上皮细胞经PTH+25(OH)D3处理,再与成骨细胞共培养)、④抑制共培养组(肾小管上皮细胞经137Cs-γ射线照射+25(OH)D3处理,再与成骨细胞共培养);并用RT-PCR方法检测共培养体系中成骨细胞的BGP、ALP、ColI、RANKL与OPG mRNA表达。结果共培养组成骨细胞的骨形成相关基因BGP、ALP、Col I mRNA表达较单培养组显著降低(P<0．001);刺激共培养组各基因表达较共培养组显著上升(P<0．05-P<0．001);抑制共培养组ALP表达水平较共培养组显著降低(P<0．001),BGP与Col I mRNA表达无显著变化。共培养组与单培养组比较,RANKL与OPC;mRNA表达显著降低(P<0．001),但RANKL／OPG比值显著高于后者(P<0．01);刺激共培养组与共培养组比较,OPG mRNA表达明显上调(P<0．01), RANKL mRNA表达变化无显著意义。RANKL／OPG比值显著降低(P<0．05);抑制共培养组与共培养组比较, OPG表达显著下降(P<0．01),RANKL/OPG比值则显著增高(P<0．01)。结论培养的近端肾小管上皮细胞对成骨细胞骨代谢相关基因表达具有直接的调节作用。建立的共培养体系可用于骨代谢相关药物筛选及作用机制研究。更多还原

【Abstract】 Purpose To investigate the direct accommodation of the proximal tubular epithelial cells (PTECs) on correlate gene expression in osteoblasts (OB). Methods The rat proximal tubular epithelial cells and osteoblasts were culture to establish the co-culture system in vitro. The PTECs were treated with PTH,γ-ray and 25(OH)D3 in advance, then co-cultured with osteoblast assigned to 4 group as follows:(1) Single- culture group (OB);(2) Co-culture group (PTECs + OB);(3) Stimulation co-culture group (PTECs stimulated by PTH + 25(()H)D, + OB) and (4) Inhibition co-culture group (PTECs irradiated byγ-rays + 25(OH)D3 + OB). The expression level of ALP, BGP, COL I , RANKL and OPG mRNA in osteoblasts were detected by RT-PCR technigne. Results The expression levels of ALP, BGP, COL I mRNA of the co-culture group were significantly lower than those in single-culture group (P<0. 001); The expression levels of ALP,BGP,COL I mRNA the stimulation co-culture group were significantly higher than those in the co-culture group (P<0. 05-P<0. 001 ) ; The expression of ALP mRNA of the inhibition co-culture group was significantly lower than that of the co-culture group (P<0. 001 ). The expression of BGP and COL I mRNA were not changed; The co-culture group compared with Single-culture group were detected that the expression levels of RANKL and OPGmRNA were significantly decreased (P<0. 001) , but the RANKL/ OPG was higher than the latter (P<0. 01). The stimulationco-culture group compared with co-culture group were detected that the expression level of OPGmRNA was remarkably up-regulated(P<0. 01), the change of RANKL mRNA was not marked, and the RANKL/ OPG was obviously decreased (P<0. 01). Compared with co-culture group, the expression of OPG mRNA of the Inhibition co-culture group was lower (P<0. 001), and RANKL was not changed, RANKL/ OPG was increased obviously (P<0. 001). Conclusions PTECs can direct regulate the gene expression of osteoblasts in vitro. The established co-culture system can be used to study the drug screening and action mechanism on bone metabolism.更多还原