摘要
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.
作者
吴飞
焦传杰
杨越
刘丰
孙志博
WU Fei;JIAO Chuan-jie;YANG Yue;LIU Feng;SUN Zhi-bo(Department of Orthopedics,Renmin Hospital,Wuhan University,Wuhan,430060,China;Department of Orthopedics,Yangxin County People’s Hospital,Hubei Yangxin,435209,China)
出处
《神经损伤与功能重建》
2020年第12期703-705,709,共4页
Neural Injury and Functional Reconstruction
基金
国家自然科学基金(No.81301052)
中央高校基本科研业务费专项资金(No.2042020kf0054,2042020gf0010)。