Yu Bin-Bin, Tong Xu-Hui, Dong Shu-Ying, Gu Yu-Chen, Jiao Hao, Ji Jie, Qu Biao
Zhonghua Nan Ke Xue. 2014 May;20(5):400-4.
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.
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.
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 microg/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).
TFLC increases the Cx43 expression and GJ function in normal TM3 Leydig cells, and the enhancement of GJ function reduces the cytotoxicity of OHP.
研究毛叶木姜子总黄酮（TFLC）对TM3睾丸间质细胞间隙连接（GJ）细胞间通讯的影响，以及TFLC在体外是否能降低奥沙利铂（OHP）的细胞毒性。
采用降落伞试验检测TFLC对体外培养的TM3细胞染料扩散的影响，通过蛋白质免疫印迹法观察TFLC处理的TM3细胞中连接蛋白43（Cx43）总蛋白表达的变化，利用免疫荧光法检测TFLC对TM3细胞膜上Cx43表达的影响，采用MTT法测定TFLC对OHP细胞毒性的影响。
在TM3细胞中，随着TFLC浓度的增加，TFLC明显增强了GJ功能。蛋白质免疫印迹法和免疫荧光法证实，TFLC显著增强了TM3细胞中Cx43总蛋白的表达及TM3细胞膜上Cx43的表达。MTT法显示，在高细胞密度（汇合形成GJ）时，20μg/ml TFLC增强了TM3细胞的GJ功能并降低了OHP的细胞毒性（P<0.05），而在低细胞密度（未汇合无GJ形成）时，TFLC对OHP的细胞毒性无影响（P>0.05）。
TFLC增加正常TM3间质细胞中Cx43的表达和GJ功能，GJ功能的增强降低了OHP的细胞毒性。
