论文部分内容阅读
该文探讨了HIF-1α修饰的大鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)对缺氧损伤所致神经元样PC12细胞凋亡的作用及其可能机制。PC12细胞分别与MSCs、HIF-1α-MSCs细胞在缺氧环境下体外共培养,分为正常组、缺氧组、MSCs组及HIF-1α-MSCs组。MTT检测不同缺氧时间PC12细胞活性。HE染色观察PC12细胞形态。Hoechst 33258染色与Annexin V-FITC双染色法检测PC12细胞凋亡。RT-PCR及细胞免疫荧光检测caspase-3的m RNA及蛋白质水平。MTT结果显示,PC12细胞活性随着缺氧时间而下降,缺氧12 h细胞活性下降至45.1%。与正常组相比,缺氧组PC12细胞凋亡率明显增高,caspase-3的m RNA和蛋白质水平明显上调,两者有显著差异(P<0.05)。与缺氧组相比,MSCs组及HIF-1α-MSCs组的凋亡率降低(P<0.05),caspase-3的m RNA和蛋白质水平下调(P<0.05),且HIF-1α-MSCs组更为明显(P<0.05)。结果证明,HIF-1α修饰的MSCs能增强MSCs的抗PC12缺氧损伤作用,其机制可能与HIF-1α-MSCs降低PC12细胞caspase-3基因表达有关。
This study explored the effect of mesenchymal stem cells (HMSCs) modified by HIF-1α on apoptosis of neuron-like PC12 cells induced by hypoxia and its possible mechanism. PC12 cells were co-cultured with MSCs and HIF-1α-MSCs in hypoxia environment respectively and divided into normal group, hypoxia group, MSCs group and HIF-1α-MSCs group. MTT assay of PC12 cells in different hypoxia time. The morphology of PC12 cells was observed by HE staining. PC12 cell apoptosis was detected by Hoechst 33258 staining and Annexin V-FITC double staining. The m RNA and protein levels of caspase-3 were detected by RT-PCR and immunofluorescence. The results of MTT showed that the activity of PC12 cells decreased with time of hypoxia, and the cell viability decreased to 45.1% at 12 h of hypoxia. Compared with the normal group, the apoptotic rate of PC12 cells was significantly increased in hypoxia group, and the m RNA and protein levels of caspase-3 were significantly up-regulated (P <0.05). Compared with hypoxia group, the apoptosis rate of MSCs group and HIF-1α-MSCs group decreased (P <0.05), and the mRNA and protein levels of caspase-3 decreased (P <0.05) More obvious (P <0.05). The results demonstrate that HIF-1α-modified MSCs can enhance the anti-PC12 effect of MSCs against hypoxia, which may be related to the decrease of caspase-3 gene expression in PC12 cells induced by HIF-1α-MSCs.