补肾益智方对AD细胞模型钙离子内流、细胞骨架、细胞凋亡的影响

Effects of Serum With Bu-Shen-Yi-Zhi Decoction on Introcellar Ca^(2+),Procedure of Cytoskeleton Reconstruction and Cell Apoptosis in AD Cellular Model

目的 :观察补肾益智方含药血清对AD(Alzheimer’sDisease)细胞模型钙离子内流、细胞骨架、细胞凋亡的影响。方法 :在神经母细胞瘤和神经胶质瘤杂交的NG10 8 15细胞培养液中加入预先老化 4d的Aβ2 5 35片段共同孵育 ,建立AD细胞模型。分别加入正常老年大鼠血清、补肾益智方高剂量含药血清和补肾益智方低剂量含药血清培养 4 8h后 ;应用激光共聚焦显微镜检测细胞内钙离子浓度 (Ca2 + )的变化情况 ;荧光显微镜观察细胞骨架的重组情况 ,流式细胞仪检测细胞的凋亡细胞。结果 :与正常大鼠血清加Aβ2 5 3 5组比较 ,高剂量补肾方含药血清组、低剂量补肾方含药血清组细胞内Ca2 + 浓度的相对荧光值明显降低 (P <0 0 1) ,而他们间两两比较则差异显著 (P >0 0 5 )。正常大鼠血清加Aβ2 5 3 5的NG10 8 15细胞组可以看到 ,细胞骨架严重解聚并向边缘扩散 ,细胞形态变性 ,还有细胞死亡现象 ;而高、低剂量补肾方含药血清组细胞骨架呈现微弱解聚现象 ,细胞形态正常 ,无变性和死亡现象。与正常大鼠血清加Aβ2 5 3 5组比较 ,高、低剂量补肾方含药血清组NG10 8 15细胞凋亡率明显减少 (P <0 0 1)。结论 :补肾益智方含药血清可以抑制细胞Ca2 + 内流 ,平衡控制细胞内Ca2 + 浓度 ;对Aβ引起NG10 8 15细胞骨架解聚、细胞变性等?

Objective:To observe the effects of serum with Bu Shen Yi Zhi Decoction on Introcellar Ca 2+ ,procedure of cytoskeleton reconstruction and cell apoptosis in AD cellular Model.Method:NG108 15 neuroblastoma x glioma hybrid cells were cultured with 5μmol olded Aβ 25 35 for establishing AD cell model.Adding 5% control rat serum,rat serum containing BSYZD(high dose)and rat serum containing BSYZD(low dose)respectively to cultured cells for 48h.Introcellar Ca 2+ was detected with an epifluorescence microscope,procedure of cytoskeleton reconstruction was obsvered with a fluorescence microscope,cell apoptosis was detected with flow cytometry.The results:The concentrations of introcellar Ca 2+ were significantly decreased in cells treatment with 5% rat serum containing BSYZD(high dose)and rat serum containing BSYZD(low dose) than the cells treatment with control rat serum (AD model).The cytoskeleton construction was distracted,the cells was deformed and death in cell treatment with control rat serum,but not in cell treatment with rat serum containing BSYZD(high dose) and rat serum containing BSYZD(low dose).A significantly higher rate of apoptosis in cells treatment with control rat serum than with rat serum containing BSYZD(high dose)and rat serum containing BSYZD(low dose).Conclusion:The serum with Bu Shen Yi Zhi Decoction could improve the introcellar Ca 2+ ,procedure of cytoskeleton reconstruction and protective the cell apoptosis in AD cellular model.