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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 regulation effect of Bushen Jianpi Fang on osteoblast cytoskeleton under micro-stress. Methods: New Zealand rabbits 18, divided into traditional Chinese medicine group, western medicine group and blank group. Twenty female SD rats were cultured. After applying the stress for 3 min and 6 min respectively, the traditional Chinese medicine group and the western medicine group were respectively cultured with the drug-containing plasma of the corresponding drugs, and the blank group was given routine culture. Laser confocal microscopy was used to detect the morphological changes and changes of fluorescence intensity. RESULTS: After stress was applied, the morphology of cells in each group changed significantly. The cytoskeleton was disaggregated and rearranged, the micro-filaments gathered around the perinuclear area, and the microtubules ruptured, shortened and bent. 6 min stress the most obvious. After 3 and 6 min of stress, there was no significant change in the fluorescence intensity at 1 h and 6 h, but after 24 h, the fluorescence intensity of the traditional Chinese medicine group increased, compared with the blank group and the western medicine group Significant significance (P <0.05). Western medicine group under 3 min stress, the fluorescence intensity of cultured 6 h was significantly higher than 1 h (P <0.05), but no significant change compared with 24 h. However, under the stress of 6 min, there was no significant change in the fluorescence intensity of each period. Conclusion: The stress applied to the cells caused some damage, the longer the stress, the more serious damage to the cells. Bushen Jianpi Chinese medicine can repair cell damage and promote the reorganization of cytoskeleton, but its role in a longer duration.