脉冲电磁场对去势大鼠骨髓源细胞体外培养破骨细胞变化的影响

Effect of pulsed electromagnetic field on the changes of osteoclasts in ovariectomized rats bone marrow culture in vitro

目的:观察脉冲电磁场对去势大鼠骨髓源性破骨细胞样细胞(OLC)形成变化及凋亡的影响。方法:30只健康3月龄雌性Wistar大鼠,随机分为A组(双侧卵巢切除+磁场治疗组、18只)、B组(双侧卵巢切除组、6只)和C组(伪去势组、6只)。A组又随机均分3组:A1、A2、A3组,电磁场频率分别为1.5、2、75Hz。术后单纯喂养3个月后对各组大鼠骨髓细胞进行培养,培养2d后,A1、A2、A3组置脉冲电磁场治疗,1次/d,每次30min,共治疗2周。之后各组培养细胞作瑞氏-吉姆萨、抗酒石酸酸性磷酸酶(TRAP)和Hoechst 33258染色,观察破骨细胞变化并计数。结果:大鼠骨髓体外培养细胞TRAP染色结果显示,C组的破骨细胞形成数量最少,其次为A2、A3、A1、B组。B组的破骨细胞形成数量与C组和A2组比较显著增多(P<0.01),B组的破骨细胞形成数量与A1、A3组比较明显增多(P<0.05)。Hoechst 33258染色结果显示,C组破骨样细胞凋亡出现数量较多,C、A2组凋亡数明显多于B组(P<0.05)。结论:脉冲电磁场能使大鼠骨髓体外培养破骨细胞形成数量减少、凋亡数增加,频率为2Hz的效果最佳,表明脉冲电磁场可能将是骨质疏松症治疗的新方法。

Objective：To investigate the effect of pulsed electromagnetic fields （PEMs） on inducing osteoclastic like cell （OLC） formation changes and apoptosis in ovariectomized （OVX） rats bone marrow culture in vitro. Methods：Thirty healthy three month old female Wistar rats were either sham operated（Sham） or ovariectomized（OVX）and randomly divided into three groups：group A （OVX＋PEMs,18 rats）,group B （OVX,6 rats） and group C （Sham,6 rats）;group A was again randomly divided into three groups：A1、A2、A3. The frequencies adopted were 1.5,2,75 Hz and 30 minutes for once a day. All rats were fed with normal diet for 3 months,then the bone marrow of all rats were cultured,2 days later,group A cells （including group A1,A2,A3） were collected and exposed to different frequencies PEMs for 2 weeks （30 min / day）. In order to observe the changes of osteoclasts and count their numbers,cells were taken for Wright Giemsa staining,tartrate resistance acid phosphatase（TRAP） staining and Hoechst 33258 staining. Results：TRAP staining results indicated the number of OLC in group C was the least,then was group A2,A3,A1,B. The number of OLC in group B was remarkly increased（P0.01;vs group C,A2）. The number of OLC in group B was significantly increased（P0.05;vs group A1,A3）. Hoechst 33258 staining results indicated the number of apoptosis of OLC in group C was more than other groups,which of group C,A2 was significantly increased （P 0.05;vs group B）. Conclusion：PEMs had decreased the formation of OLC and increased the number of apoptosis of OLC in ovariectomized （OVX） rats bone marrow culture in vitro,the effects of 2 Hz was the best. PEMs would be a new way of osteoporosis therapy.