缝隙连接细胞间通讯在丙戊酸诱导大鼠脂肪源干细胞向许旺细胞分化中的作用

Effect of Gap Junction Intercellular Communication on Valproic Acid-Induced Differentiation of Adipose Tissue-derived Stem Cells into Schwann Cells

目的:探讨缝隙连接细胞间通讯在丙戊酸诱导大鼠脂肪源干细胞(ADSCs)向许旺细胞分化中的作用。方法:体外分离培养大鼠ADSCs并传至第3代,将ADSCs分别用不含丙戊酸(A组)、含1.0 mmol/L丙戊酸(B组)、1.0 mmol/L丙戊酸+2.5μmol/L油酸酰胺(C组)的诱导液培养。显微镜下观察细胞形态变化,分别用免疫细胞化学法和实时荧光定量PCR法检测各组中枢神经组织蛋白S100、神经元特异性烯醇化酶(NSE)、巢蛋白(Nestin)、缝隙连接蛋白43(Cx43)的蛋白表达水平和mRNA转录水平。结果:B、C组S100、NSE、Nestin蛋白表达水平和mRNA转录水平均显著高于A组(P<0.01),C组低于B组(P<0.01);B、C组Cx43蛋白表达水平和mRNA转录水平均显著高于A组,B、C组之间差异无统计学意义(P>0.05)。结论:Cx43介导的缝隙连接细胞间通讯在丙戊酸诱导ADSCs向许旺细胞分化的过程中可能有重要作用。

Objective:The aim of this study was to investigate the effect of gap junction intercellular communication on the valproic acid-induced differentiation of adipose tissue-derived stem cells(ADSCs)into Schwann cells.Methods:ADSCs from rats were isolated and cultured.At passage 3,the cells were inoculated with medium containing no valproic acid(group A),1.0 mmol/L valproic acid(group B),or 1.0 mmol/L valproic acid+2.5μmol/L oleamide(group C).Changes in cell morphology were observed by microscopy.Immunocytochemical staining and real-time quantitative PCR were employed to assess the protein expression and mRNA transcription levels of S100,neuron-specific enolase(NSE),Nestin,and Connexin-43(Cx43).Results:Compared to group A,groups B and C showed a significant increase in the protein expression and mRNA transcription levels of S100,NSE,and Nestin(P<0.01),and group C showed lower levels than group B(P<0.01).Groups B and C also showed significantly higher levels of protein expression and mRNA transcription of Cx43 than group A,and there was no significant difference between group B and group C(P>0.05).Conclusion:Cx43-mediated gap junction intercellular communication may serve an important role in the valproic acid-induced differentiation of ADSCs into Schwann cells.