褪黑激素对犬脂肪干细胞向雪旺细胞分化过程中凋亡发生的抑制作用研究

Inhibitory effect of melatonin on apoptosis in the process of differentiation of canine adipose stem cells into schwann cells

为了探讨褪黑激素(Mel)对犬脂肪干细胞(ADSCs)向雪旺细胞(SCs)诱导分化过程中凋亡发生的影响。本实验将犬ADSCs诱导分化为SCs,并使用免疫荧光法进行鉴定。采用CCK8法选取Mel的最佳浓度。应用q RT-PCR法和Western-blot法检测无Mel组和Mel处理组中ADSCs向SCs诱导过程中细胞凋亡相关基因Caspase-3、Caspase-8、Bcl-2和Bax的m RNA和蛋白表达水平。结果显示,50 nmol/L Mel处理过的细胞活力最高;ADSCs向SCs诱导后S-100和GFAP呈阳性表达,并且观察到Mel的添加并不会影响到SCs的表型特征。ADSCs向SCs诱导后Bcl-2表达量明显低于对照组,Caspase-3、Caspase-8和Bax的表达量显著高于对照组,细胞凋亡较多;在加入Mel后Bcl-2表达量显著升高,Caspase-3、Caspase-8和Bax的表达量显著下降。结果说明Mel可以显著抑制犬ADSCs向SCs诱导分化过程中的凋亡发生。

In order to investigate the effect of melatonin(Mel)on apoptosis in the process of differentiation of canine adipose stem cells(ADSCs)into schwann cells(SCs).Canine ADSCs were induced to differentiate into SCs and identified by immunofluorescence.The optimum concentration of Mel was selected by the CCK8 method.Moreover,q RT-PCR and Western-blot were used to detect the m RNA and protein expression levels of apoptosis related genes(Caspase-3、Caspase-8、Bcl-2、Bax)during ADSCs induction into SCs in the Mel-free group and the Mel-treated group.The results showed that the cells treated with Mel at 50 nmol/L had the highest activity.After induction of ADSCs into SCs,S-100 and GFAP showed positive expression,and it was observed that the addition of Mel did not affect the phenotypic characteristics of SCs.Moreover,when ADSCs were induced to SCs,the expression of Bcl-2 was significantly lower than that of the control group,and the expression of Caspase-3,Caspase-8 and Bax was significantly higher than that of the control group,with more apoptosis.After the addition of Mel,the expression of Bcl-2 increased significantly,while the expression of Caspase-3,Caspase-8 and Bax decreased significantly.These results suggest that Mel can significantly inhibit apoptosis in the process of differentiation of canine ADSCs into SCs.