许旺细胞提取与纯化的新方法

背景:幼年动物脂肪组织中前脂肪细胞的生物学行为与肥胖的发病和防治关系密切,而采用细胞生物力学实验方法直接观测力学振荡对前脂肪细胞生物力学行为变化可为推拿按摩治疗单纯性肥胖原理研究提供更直接的实验证据.目的:通过对体外培养的幼鼠前脂肪细胞实施不同频率的机械力刺激,观测细胞活力、增殖与细胞凋亡的变化.方法:体外培养SD幼鼠前脂肪细胞.对细胞进行鉴定后,通过细胞恒温培养振荡器对前脂肪细胞实施动态机械力学刺激.根据不同处理频率分别分为0(空白对照).1.5和3 Hz共3个组,每组振荡时间均为30 min,每12 h振荡1次,共处理3 d.观察细胞活力、增殖与凋亡率的变化.结果与结论:培养初期细胞形态似成纤维细胞,油红O染色后,细胞内出现红染颗粒,证明体外培养的幼鼠细胞经鉴定为前脂肪细胞.随实验振荡力的刺激,前脂肪细胞的活力和增殖有显著抑制(P<0.05或P<0.01),但力学振荡刺激对前脂肪细胞的凋亡与空白对照组比较差异无显著性意义(P>0.05).结果提示推拿防治青少年单纯性肥胖的细胞生物学机制可能主要是通过抑制前脂肪细胞的活力和增殖而作用的.

Preadipocyte viability, proliferation, and apoptosis in young rats following dynamic mechanical force stimulation

BACKGROUND: The biological behaviors of preadipocytes in adipose tissue of young animals have been closely linked to the onset and prevention and treatment of obesity. Observing mechanical oscillation effects on biomechanical behaviors of preadipocytes using biomechanical stimuli would provide more direct experimental evidence for treatment of simple obesity using manipulation and massage therapy.OBJECTIVE: Different frequencies of mechanical force stimulation were performed on preadipocytes from young rats cultured in vitro to observe the changes in cell viability, proliferation, and apoptosis.METHODS: Preadipocytes from SD young rats were in vitro cultured. Following identification, preadipocytes were dynamically, mechanically stimulated through the use of constant temperature oscillator. According to different treatment frequencies, three groups were set: 0 Hz (blank control), 1.5 Hz, and 3 Hz. A 30-minute oscillation, once every 12 hours, total 3 days, was performed in each group. Following dynamic mechanical stimulation, cell viability, proliferation, and apoptosis in young rats were observed. RESULTS AND CONCLUSION: In the initial stage of culture,cells exhibited the morphology similar to fibroblasts. After oil red O staining, red particles appeared in the cells, indicating that the young mouse cells cultured in vitro were preadipocytes. With stimulation of oscillation force, the viability and proliferation of preadipocytes were significantly inhibited (P < 0.05 or P < 0.01). There wasno significant difference in effects of mechanical oscillation on preadipocyte apoptosis between 0 Hz and 1.5 Hz, 3 Hz groups (P > 0.05). These findings indicate that thecell biological mechanism underlying preventing simple obesity in adolescents is to inhibit the viability and proliferation of preadipocytes