论文部分内容阅读
目的:探讨豹皮樟总黄酮(TFLC)对体外培养的小鼠睾丸Leydig细胞系(TM3)缝隙连接(GJ)功能的作用,以及TFLC是否减轻化疗药奥沙利铂(OHP)的细胞毒作用。方法:荧光示踪法测定体外培养的TM3细胞之间荧光传递,反映GJ的功能;Western印迹法检测TFLC对连接蛋白43(Cx43)表达的影响;细胞免疫荧光法观察TFLC对TM3细胞胞膜Cx43蛋白表达的影响;MTT法检测OHP的细胞毒作用,以及TFLC对OHP细胞毒的影响。结果:在TM3细胞中GJ的功能随着TFLC浓度的增加而增强;Western印迹和免疫荧光实验结果显示TFLC可显著增强TM3细胞中Cx43蛋白和膜Cx43蛋白的表达;MTT结果表明,高密度接种细胞(生长融合,有GJ形成),20μg/ml TFLC增强细胞GJ功能,且减轻OHP的细胞毒作用[细胞存活率(89.5±1.8)%高于单用ONP组(80.0±1.5)%,(P<0.05)];低密度接种细胞(生长未融合,无GJ形成),TFLC对OHP的细胞毒作用没有影响(P>0.05)。结论:TFLC增强睾丸正常细胞TM3中Cx43的表达,并增强细胞间的GJ功能;TFLC减轻OHP在TM3细胞的毒性作用,并可能与增强GJ功能有关。
AIM: To investigate the effect of TFLC on the gap junction (GJ) function of cultured mouse Leydig cell line (TM3) in vitro and the effect of TFLC on cytotoxicity of oxaliplatin (OHP) . Methods: Fluorescence tracer method was used to determine the fluorescence transmission between TM3 cells in vitro and the function of GJ. Western blotting was used to detect the effect of TFLC on the expression of connexin43 (Cx43). The effect of TFLC on Cx43 Protein expression; MTT assay cytotoxicity of OHP, and TFLC OHP cytotoxic effect. Results: The function of GJ in TM3 cells increased with the increase of TFLC concentration. Western blotting and immunofluorescence assay showed that TFLC significantly enhanced the expression of Cx43 and Cx43 in TM3 cells. The results of MTT assay showed that high-density seeding cells (GJ formation), 20μg / ml TFLC enhanced the function of GJ cells and reduced the cytotoxicity of OHP [cell survival rate (89.5 ± 1.8%] was higher than that of ONP group (P (80.0 ± 1.5)% <0.05)]. Low-density inoculated cells (unfused without GJ formation), TFLC had no effect on the cytotoxicity of OHP (P> 0.05). Conclusion: TFLC enhances the expression of Cx43 and enhances the GJ function in TM3 cells. TFLC attenuates the toxicity of OHP in TM3 cells and may be related to the enhancement of GJ function.