人甲状旁腺激素1-34对培养的人牙髓细胞PTH-PTHrP受体mRNA表达的影响

Effect of human parathyroid hormone 1-34 on the expression of mRNA of PTH/PTHrP receptor in human dental pulp cells cultured in vitro

目的:观察人甲状旁腺激素1-34(hum an Parathyroid hormone 1-34,hPTH1-34)对PTH-PTHrP(Parathyroid hormone-Parathyroid hormone related prote in)受体mRNA在体外培养的人牙髓细胞中表达的影响,探讨PTH影响牙本质形成和矿化的机制。方法:将培养的第4代牙髓细胞分为两组:实验组培养基加入含33.3 nmol/L hPTH1-34,在其培养的第5,10,15,20天提取总RNA,以RNA为模板在逆转录酶作用下合成cDNA,加入特异引物进行PCR扩增,同时扩增β-肌动蛋白的cDNA作为半定量RT-PCR的内参照,对PCR产物进行半定量分析。结果:在人牙髓细胞培养的5、10 d,对照组和实验组的PTH-PTHrP受体mRNA水平均较低,10 d以后PTH-PTHrP受体mRNA水平开始升高,实验组高于对照组,差异有统计学意义。结论:PTH-PTHrP受体的表达与牙髓细胞的分化密切相关;hPTH1-34可以促进PTH-PTHrP受体的表达,PTH可能通过促进PTH-PTHrP受体的表达或与PTH-PTHrP受体结合影响牙本质的形成和矿化。

AIM： To study the change of expression of the PTH/PTHrP receptor in human dental pulp cells cultured in vitro. METHODS： The fourth passage of human dental pulp cells were cultured up to 5,10,15,20 days. The PTH group was cultured in the medium containing 33.3nmol/L hPTH1 - 34. PTH/PTHrP receptorg mRNA were enlarged by RT - PCR. The absorbence were measured and compared with β - actin. RESULTS： The mRNA of PTHrP receptor expressed lower when the cells were cultured in 5 and 10 days in both the experimental group and control group . But after 10 days, the expressions all increased with the cultured time. But the expression is higher in the experimental group than that in the control group. CONCLUSION： PTH is related with the differentiation Of pulp cells, hPTH1 -34 may promote the expression of PTH/PTHrP receptor. The formation and mineralization of dentin may be effected by parathyroid hormone itself or by combining with PTH/PTHrP receptor.