补肾活血固齿方对MC3T3-E1细胞BMP2表达及TRPV5/TRPV6通道影响的实验研究

The experimental study of effects of Bushen Huoxue Guchi formula on BMP2 and TRPV5/TRPV6 channels expressions in MC3T3-E1 cells

目的观察补肾活血固齿方对小鼠颅顶前成骨细胞(MC3T_3-E1细胞)骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)表达及TRPV5/TRPV6通道的影响,探讨补肾活血固齿方成骨的作用机制。方法按体表面积折算等效剂量的补肾活血固齿方灌胃大鼠,制备含药血清;等剂量生理盐水灌胃大鼠,制备无药血清作为对照。用10%含药血清、无药血清分别培养MC3T_3-E1细胞24、48、72h,RT-PCR法检测MC3T_3-E1细胞BMP2、TRPV5/TRPV6mRNA的表达。结果含药血清组MC3T_3-E1细胞的BMP2、TRPV6mRNA表达量较无药血清组增加(P<0.05),且随时间延长有逐渐增加趋势,但差异无统计学意义(P>0.05)。含药血清组和无药血清组MC3T_3-E1细胞中均未检测到TRPV5 mRNA的表达。结论补肾活血固齿方可能是通过钙离子通道TRPV6上调BMP2的表达、诱导成骨细胞分化的。

Objective To observe the effect of Bushen Huoxue Guchi Formula on bone morphogenetic protein 2（BMP2）and TRPV5/TRPV6 channel expressions,and to explore the mechanism of Bushen Huoxue Guchi Formula in bone formation.Methods Herb-contained serum was prepared from rats by gavage with Bushen Huoxue Guchi Formula by using body surface area converted equivalent dose.Blank-contained serum as control group was prepared by gavage with normal saline.The BMP2 and TRPV5/TRPV6 mRNA expressions were detected in MC3T3-E1 cells by RT-PCR after the cells were treated with 10% herb-contained serum or blank-contained serum for 24 h,48hand 72 h,respectively.Results BMP2,TRPV6 mRNA were significantly increased in herb-contained serum treatment group compared with blankcontained serum treatment group（P〈0.05）. With the prolongation of time,the mRNA expression of BMP2 and TRPV6 had a gragually increased tendency,but there was no statistically significant difference（P〉0.05）.TRPV5 mRNA was not detected either in herbcontained serum treatment group or blank-contained serum treatment group.Conclusion Bushen Huoxue Guchi Formula may upregulate BMP2 expression through TRPV6 channels to induce differentiation of osteoblast.