Total flavonoids of litsea coreana decreases the cytotoxicity of oxaliplatin in TM3 Leydig cells via enhancing the function of gap junction

Objective: To investigate the effects of total flavonoids of Litsea Coreana( TFLC) on the gap junction( GJ) intercellular communication in TM3 testicular Leydig cells and whether TFLC can reduce the cytotoxicity of oxaliplatin( OHP) in vitro. Methods: We detected the effect of TFLC on the dye spread of the in vitro cultured TM3 cells by parachute assay,observed changes in the expression of connexin 43( Cx43) total protein in the TFLC-treated TM3 cells by Western blot,and determined the effects of TFLC on the expression of Cx43 on the membrane of the TM3 cells by immunofluorescence assay and on the cytotoxicity of OHP by MTT assay. Results: TFLC obviously enhanced the GJ function with the increasing of the TFLC concentration in the TM3 cells. Western blot and immunofluorescence assay confirmed that TFLC significantly enhanced the expression of Cx43 total protein and Cx43 expression on the membrane of the TM3 cells. MTT assay showed that at a high cell density( confluent with GJ formation),20 μg / ml TFLC enhanced the GJ function of the TM3 cells and reduced the cytotoxicity of OHP( P 0. 05),while at a low density( preconfluent with no GJ formation),TFLC exhibited no effect on the cytotoxicity of OHP( P 0. 05). Conclusion: TFLC increases the Cx43 expression and GJ function in normal TM3 Leydig cells,and the enhancement of GJ function reduces the cytotoxicity of OHP.