目的:研究谷氨酸(Glu)对大鼠脂肪干细胞Adipose-derived stem cells(ADCSs)诱导分化神经细胞的作用及机制。方法:取成年大鼠腹股沟脂肪组织进行体外细胞培养,采用免疫组化方法检测证实为ADSCs。对照组为正常培养的ADSCs并诱导分化神经细胞,谷氨酸(Glu)处理组加入不同浓度的Glu,MTT比色法观察脂肪干细胞的存活率。结果:从ADSCs诱导分化的细胞包括神经元及神经胶质细胞,免疫组化结果显示神经元特异性烯醇化酶(NSE)染色阳性和胶质纤维酸性蛋白(GFAP)染色阳性。Glu处理组给药24h后,与对照组比较ADSCs存活率明显降低,50μmol·L^-1Glu组细胞存活率为83.98%,与对照组比较差异无显著性(P〉0.05);100及1000μmol·L^-1Glu组干细胞存活率(分别为66.82%和17.08%)低于对照组(P〈0.05,P〈0.01)。结论:Glu对ADSCs有损伤作用,随着Glu剂量的增加,ADSCs的存活率逐渐降低,二者呈剂量依赖关系。
Objective: To explore the influence and mechanism of the ADSCs cultivated in vitro induced by glutamate of rats. Methods: The adult rat inguinal region tissues were cultivated in vitro and the cells were confirmed as the ADSCs with immunohistochemical method. The ADSCs cultured normally were used as control, the different concent rations of glutamate were added, then the survival rate of ADSCs was observed with MTT colorimetry. Results: The differentiation cells from ADSCs had both neurons and neuroglial cellsthat presented neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) dying masculinere spectively. After treated with glutamate for 24 h , the survival rates of ADSCs in glutamate groups were obviously reduced compared with control group. The survival rate in 50 μmol·L^-1 glutamate group was 83.98 % ,compared with control group , there was no significant difference (P〈0. 05) . The survival rates of the ADSCs in 100 and 1 000 μmol·L^-1 glutamate groups (66.82%, 17.08%) were significantly lower than that in control group ( P〈0.05 , P〈0.01). Conclusion: Glutamate has function of damaging on the ADSCs in a dose-dependent manner ; when the dose of glutamate increases , the survival rate of the ADSCs decreases gradually.