Abstract: Objective To evaluate the effects of three antidepressants with different mechanisms on connexin expression and channel function of hippocampal astrocytes. Methods The cytotoxicity of antidepressants with different mechanisms on hippocampal astrocytes was assessed by CCK-8 assay. The effects of three antidepressants with different mechanisms on gap junction function and the expression level of connexin 43 protein （Cx43） were determined by cell inoculation fluorescence tracing and western blot. Results CCK-8 assay showed that 25.00 µmol/L fluoxetine, 0.10 µmol/L amitriptyline and 0.20 nmol/L venlafaxine had no cytotoxicity on hippocampal astrocytes （all P > 0.05）. In addition, 25.00 µmol/L fluoxetine, 0.10 µmol/L amitriptyline and 0.20 nmol/L venlafaxine could significantly inhibit the fluorescence transfer function of gap junction in hippocampal astrocytes （all P < 0.05）, whereas three antidepressants exerted on effects upon the expression of Cx43 in hippocampal astrocytes （all P > 0.05）. Conclusion Antidepressants can significantly reduce the gap junction function composed of Cx43 in hippocampal astrocytes.