补肾宁心方药物血清对鼠成骨细胞的增殖周期与凋亡的影响

目的:探讨补肾宁心方药物血清对成骨细胞(osteob last,OB)增殖周期与凋亡的调控作用。方法:颅骨酶解法培养OB,体外模拟雌激素撤退现象,分别予以5%~30%系列浓度的对照鼠血清或中药血清培养,流式细胞仪分析DNA含量,Annexin V-FITC/PI双标记流式细胞仪分析细胞早期凋亡,透射电镜观察OB超微结构。结果:DNA含量分析显示与相应对照组相比,10%~20%药物血清组的S期和G2/M期细胞百分比明显增加,G0/G1期细胞百分比明显下降,细胞增殖指数(PI)显著增加(分别为P<0.05和P<0.01);10%~20%的药物血清还具有显著的抗凋亡作用(分别为P<0.05和P<0.01);透射电镜可见经20%药物血清诱导后的OB细胞出现典型的细胞器明显增多,内质网扩张,线粒体丰富等形态学改变。结论:补肾宁心方药物血清在体外可有效地促进鼠OB的增殖周期,从而促进OB增殖并抑制其凋亡。

Effects of Pharmacological Serum after Treated by Bushen Ningxin Decoction (BSNXD) on Proliferating Cycle and Apoptosis of Murine Osteoblasts

Objective:To investigate into the effects of the BSNXD-treated pharmacological serum on the proliferating cycle and apoptosis of osteoblasts(OBs).Methods: The OBs were isolated by the enzyme-digested assay from calvaria of neonatal BALB/c mice and then cultured.The OBs were treated in vitro with the pharmacological serum in a series of concentration.The apoptosis rate and DNA content of OBs were analyzed by flow cytometry(FCM),and their ultrastructure was evaluated by TEM.Results: The pharmacological serum in 10-20% concentration increased the S-phase and G_(2)/M phase of OB cell cycle,improved the proliferation index(PI,P<0.05,P<0.01).The pharmacological serum at the concentration of 10-20% was capable of inhibiting apoptosis of OBs(P<0.05,P<0.01).Moreover,the OBs treated by the pharmacological serum showed more organelles,expanded endoplasmic reticulum(ER) and abundant mitochondria(M).Conclusion: BSNXD improves their proliferation and inhibits apoptosis of OBs induced by serum deprivation.