补肾健脾方对微应力下成骨细胞骨架调控的影响

Regulative Effect of Bushen Jianpi Decoction on Osteoblast Cytoskeleton Under Microstress in Vitro

目的:探讨补肾健脾方对微应力下成骨细胞骨架的调控作用。方法:新西兰兔18只,分为中药组、西药组和空白组。培养细胞用雌性SD大鼠20只。分别施加3 min和6 min应力后中药组和西药组分别以相应药物的含药血浆培养,空白组给予常规培养。采用激光共聚焦显微镜检测细胞的形态学变化及荧光强度变化。结果:施加应力后,各组细胞形态发生明显变化,出现细胞骨架发生解聚和重排,微丝向核周集聚,微管发生断裂,变短,弯曲。以6 min应力最为明显。中药组在3 min和6 min应力下,其培养1 h和6 h后的荧光强度无明显变化,但经过24 h培养后,中药组的荧光强度增强,与空白组、西药组比较,差异有显著性意义(P<0.05)。西药组在3 min应力下,其培养6 h的荧光强度明显高于1 h(P<0.05),但与24 h比较无明显变化。但在6 min应力下,各个时段的荧光强度无明显变化。结论:施加的应力对细胞造成一定的损伤,应力时间越长,其对细胞的损伤越严重。补肾健脾中药能修复细胞的损伤,促进细胞骨架的重组,但其起效作用时间较长。

Objective： To investigate the protective effect of Bushen Jianpi decoction on osteoblast cytoskeleton in vitro under microstress. Methods： Eighteen New Zealand rabbits were divided into the blank group, western medicine group and Chinese medicine group. Primary cultured calvaria osteoblasts （OB） were obtained from 20 female SD rats. After given mechanical strain for 3-min and 6-min, OB of western medicine group and Chinese medicine group were cultured with the corresponding medicated serum. OB of the blank group were cultured by routine medium. The changes of morphology and fluorescence intensity of OB were measured with laser scanning confocal microscope. Results： After given microstress, the changes of cell morphology were obvious in each group, manifested as cytoskeleton depolymerization and rearrangement, perinuclear concentration of microfibril, and microtubule fracture, shortening, and curving. The changes were the most obvious after stress for 6 min. The fluorescence intensity of Chinese medicine group had no significant changes after culturing for one hour and 6 hours under 3-min and 6-min microstress. While after 24-hour culture, the fluorescence intensity of Chinese medicine group was significantly increased （P 〈 0.05 compared with the blank group and western medicine group）. Under 3-min microstress, the fluorescence intensity of western medicine group was significantly higher after 6-hour culture than that after one-hour culture（P 〈 0.05）, but did not differ from that after 24-hour culture. Under 6-min microstress, the fluorescence intensity of western medicine group remained unchanged after culturing for different time. Conclusion： The microstress can cause damage to OB, and the damage increases with the stress time. Bushen Jianpi decoction can repair cell damage and promote reconstruction of OB cytoskeleton, but the effect starts after a time.