大鼠心肌组织提取物抑制缺氧和无血清诱导的大鼠骨髓间充质干细胞凋亡

Tissue extracts from myocardium of rats protects bone marrow stromal cells against hypoxia and serum deprivation-induced apoptosis

目的:观察大鼠心肌组织提取物抑制以缺氧和无血清方法诱导的大鼠骨髓间充质干细胞凋亡现象并初步探讨其机制。方法:原代培养大鼠骨髓间充质干细胞,建立缺氧和无血清诱导细胞凋亡模型,超速离心法提取大鼠心肌组织提取物,以CCK8法检测缺氧和无血清条件下不同浓度心肌组织提取物作用后的细胞数量,筛选适宜提取物作用浓度;相差显微镜下观察正常对照组、凋亡模型组和心肌组织提取物组细胞凋亡情况,进一步以Hoechst染色法检测细胞核形态变化,计数染色质浓缩细胞所占比例,Western blot法检测各组caspase-3蛋白表达量。结果:随着心肌组织提取物浓度的升高,缺氧和无血清条件下细胞存活的数量也随之增加,心肌组织提取物的最佳作用浓度为100μg/mL。心肌组织提取物组、正常对照组与凋亡模型组比较,细胞凋亡数明显减少,caspase-3蛋白裂解活化程度显著降低。结论:大鼠心肌组织提取物能够抑制缺氧和无血清诱导的大鼠骨髓间充质干细胞凋亡,其浓度超过100μg/mL时抑制细胞凋亡作用明显,caspase-3参与了该现象的调控。

Objective： To explore tissue extracts from myocardium of rats protects bone marrow stromalcells （BMSC） against hypoxia/serum deprivation （SD）-induced apoptosis, and investigate its potential meclaanlsm. Methods： The BMSC were isolated from female SD rat and cultured in vitro. The ischemic microenvironment was simulated by hypoxia/SD and tissue extracts from myocardium of rats were extracted by ultracentrifugation. Cell counting kit was used after 24 h hypoxia/SD treatment to observe the cell number of BMSC and to select the proper concentration. The inverted phase contrast microscope observation and heochst 33258 nucleus staining were used to detect the morphological changes of the cells, Western blot was used to detect the expression of caspase-3. Results： The result of cell counting kit showed that with the extracts concentration increasing, the cell number increased. The proper concentration of myocardium tissue extracts was 100 μ g/mL. Compared with the apoptotic model （AM） group, there were less apoptotic cells in myocardium tissue extracts group and control group. The expression level of cleaved caspase-3 detected was higher in AM group than that in the NMTE and control group. Conclusion： Tissue extracts from myocardium of rats protect BMSC against hypoxia/SD-induced apoptosis. The protective effect is significant when the extracts concentration reached 100 μg/mL. The protective effect is related to caspase-3 signal pathway.