党参多糖对Aβ1-40诱导PC12细胞衰老的影响与机制研究

Study on effect and mechanism of Radix Codonopsis polysaccharide on PC12 cells senescence induced by Aβ1-40

目的:探讨党参多糖对Aβ1-40诱导PC12细胞衰老的延缓作用。方法:将PC12细胞分为4组:分别是正常组、Aβ1-40组、Aβ1-40+党参多糖组、党参多糖组;采用CCK-F法筛选Aβ1-40和党参多糖的最佳浓度;应用倒置显微镜观察细胞形态;采用β-半乳糖苷酶染色检测细胞衰老;应用免疫荧光法和Western Blot法检测Aβ1-40和党参多糖对衰老相关蛋白(P16、P21)的表达情况。结果:通过CCK-F法确定Aβ1-40和党参多糖的最佳浓度分别为1μmol/L和50μg/mL;与正常组比较,Aβ1-40组细胞突起减少,细胞活性下降(P<0.01),衰老细胞数量增多(P<0.01),衰老相关蛋白P16、P21表达增加(P<0.01);党参多糖组细胞形态正常,细胞互联成网状生长,细胞活力增加(P<0.01),衰老细胞数量少(P<0.01),细胞衰老相关蛋白P16、P21表达降低(P<0.01)。与Aβ1-40组比较,Aβ1-40+党参多糖组细胞突起较长,细胞活力增加(P<0.01),衰老细胞数量减少,P16、P21表达下降(P<0.01)。结论:党参多糖能通过减少衰老细胞的数量以及下调P16、P21蛋白表达,从而达到延缓Aβ1-40诱导PC12细胞衰老的作用。

Objective: To investigate the anti-aging effects of Radix Codonopsis polysaccharide on Aβ1-40 induced PC12 cells. Methods: PC12 cells were divided into four groups: normal group, Aβ1-40 group, Aβ1-40+Radix Codonopsis polysaccharide group, Radix Codonopsis polysaccharide group. CCK-F method was used to screen the best concentration of Aβ1-40 and Radix Codonopsis polysaccharide. Inverted microscope was used to observe cell morphology. β-galactosidase staining was used to detect cell senescence. Immunofluorescence and Western Blot were used to detect the expression of Aβ1-40 and Radix Codonopsis polysaccharide on aging related proteins(P16, P21). Results: The best concentrations of Aβ1-40 and Radix Codonopsis polysaccharide were 1μmol/L and 50μg/mL. Compared with the normal group, Aβ1-40 group had decreased cell processes, decreased cell activity(P<0.01), increased number of senescent cells(P<0.01), and increased expression of senescence related proteins P16 and P21(P<0.01), and the morphology of cells was normal, the cells interconnected into a network, the cell viability increased(P<0.01), the number of senescent cells was less(P<0.01), and the expression of p16 and p21 in cells was lower(P<0.01). Compared with Aβ1-40 group, Aβ1-40+Radix Codonopsis polysaccharide group had longer cell processes,increased cell viability(P<0.01), decreased number of senescent cells(P<0.01), and decreased expression of P16 and P21(P<0.01).Conclusion: Radix Codonopsis polysaccharide can delay the senescence of PC12 cells induced by Aβ1-40 by reducing the number of senescent cells and downregulate the expression of P16 and P21 proteins.