复方接骨片含药血清对BMP-2沉默的骨髓间充质干细胞体外增殖及分化影响

Effect of Compound Jiegu Tablets on the Proliferation and Differentiation of Bone Marrow Mesenechymal Stem Cells with BMP-2 Gene Silencing

目的:观察复方接骨片含药血清对BMP-2沉默的骨髓间充质干细胞体外增殖及分化的影响。方法:设计三对BMP-2 si RNA,利用脂质体介导的si RNA转染骨髓间充质干细胞,PCR及Western blot法筛选活性;将复方接骨片含药血清加入BMP-2沉默的骨髓间充质干细胞中,MTT法及碱性磷酸酶法检测细胞增殖和分化情况。结果:三对si RNA中,si RNA3在抑制BMP-2的m RNA和蛋白表达的效果最好,抑制率可达61.6%和50.9%;正常骨髓间充质干细胞加入含药血清,48 h后可明显提高增殖效率(P<0.05)及碱性磷酸酶活性(P<0.01);骨髓间充质干细胞转染BMP-2 si RNA3,48 h后碱性磷酸酶活性显著降低(P<0.01),72 h后细胞活性显著降低(P<0.01),而加入含药血清干预,可显著拮抗上述抑制作用(P<0.01)。结论:复方接骨片含药血清能有效拮抗BMP-2 si RNA对骨髓间充质干细胞体外增殖及分化的抑制作用。

Objective：To study the effect of Compound Jiegu Tablets on the proliferation and differentiation of bone marrow mesenechymal stem cells（BMSCs）with BMP-2 gene silencing. Methods：Three BMP-2 si RNA duplexes were designed,and transfected to BMSCs by liposome. One of them with higher silencing activity was screened by PCR and western blot. Serum of Compound Jiegu Tablets were added to BMSCs with BMP-2 gene silencing. Cell proliferation and differentiation were detected by MTT and AKP activity. Results：Transfection of si RNA3 significantly decreased the m RNA and protein expression of BMP-2,the inhibition rate were 61.6% and 50.9%;After the medicated serum added into the normal BMSCs for 48 h,cell proliferation and activity of AKP were significantly increased（P〈0.05 or P〈0.01）;after BMSCs transfected with si RNA3,activity of AKP and cell proliferation were significantly reduced respectively at 48 h and 72 h（P〈0.01）. The medicated serum could significantly antagonize the effect of the inhibition above（P〈0.01）. Conclusion：Compound Jiegu Tablets medicated serum could attenuate the inhibitory effect of BMP-2 si RNA and promote the proliferation and differentiation of BMSCs in vitro.