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目的建立体外骨髓基质干细胞(MSCs)与Aβ1-40损伤PC12的共育体系,探讨共育体系抑制Aβ1-40致PC12凋亡的效应与可能机制。方法分别培养MSCs与PC12,以Aβ1-40刺激PC12后,用转移筛网转移PC12。实验分A组:正常培养的PC12+MSCs共育;B组:Aβ1-40刺激的PC12+ MSCs共育;C组:正常PC12的培养上清+MSCs;D组:受损PC12上清+MSCs;E组:普通1640培养的MSCs。用PI和Annexin-V进行细胞的双染凋亡检测及电镜检测PCI2凋亡,以ELISA方法检测各组中上清液的TGF-β、NGF、BDNF、bFGF含量。结果骨髓基质干细胞与Aβ1-40损伤PC12共育组即B组凋亡细胞数最少(B组46.17%±8.28%,对照组86.39%±9.34%,F=61.637,P<0.01),ELISA结果显示各组均能检测到bFGF,B组上清中bFGF分泌最高[B组(598.76±41.32) pg/ml,对照组(296.43±47.86) pg/ml,F=24.15,P<0.01)],各组均检测到TGF-β、NGF和BDNF分泌,但差异无统计学意义。结论MSCs与Aβ1-40刺激的PC12的共育体系能减少受损PC12的凋亡。
Objective To establish a co-culture system of bone marrow stromal stem cells (MSCs) and Aβ1-40-injured PC12 in vitro and to explore the effect and possible mechanism of co-culture system on inhibiting the apoptosis of PC12 induced by Aβ1-40. Methods MSCs and PC12 were cultured respectively. After PC12 was stimulated by Aβ1-40, PC12 was transferred by transfer screen. Group A: PC12 + MSCs co-cultured in normal culture; Group B: co-cultured PC12 + MSCs stimulated by Aβ1-40; Group C: MSCs supernatant + MSCs from normal PC12; Group D: injured PC12 supernatant + MSCs; Group E: Normal 1640 cultured MSCs. Double staining and PI staining with Annexin-V were used to detect the apoptosis of the cells and the apoptosis of PCI2 was detected by electron microscopy. The contents of TGF-β, NGF, BDNF and bFGF in the supernatants were detected by ELISA. Results The number of apoptotic cells in group B was the lowest (B group 46.17% ± 8.28%, control group 86.39% ± 9.34%, F = 61.637, P <0.01). The results of ELISA BFGF was detected in each group, the highest bFGF secretion in the supernatant of group B [598.76 ± 41.32 pg / ml in control group (296.43 ± 47.86) pg / ml, F = 24.15, P <0.01) TGF-β, NGF and BDNF secretion were detected in all groups, but the difference was not statistically significant. Conclusion Co-culture of MSCs with Aβ1-40 stimulated PC12 can reduce the apoptosis of damaged PC12.