菟丝子黄酮对BPA致小鼠睾丸间质细胞分泌雄性激素异常的保护作用

Protective Effect of Flavonoids from Cuscuta chinensis on BPA-induced Abnormal Secretion of Male Hormone of LeydigCells in Mice

为研究菟丝子黄酮对双酚A(BPA)干扰小鼠睾丸间质细胞激素分泌的缓解作用,以TM3小鼠睾丸间质细胞为对象,采用1μg/mL～500μg/mL的菟丝子黄酮与TM3(小鼠睾丸间质细胞)细胞共同培养24 h,以MTT法检测细胞增殖,确定菟丝子黄酮对TM3细胞的安全剂量。然后采用1μmol/mL～200μmol/mL的双酚A干扰TM3细胞24 h,以MTT法检测细胞增殖,通过TM3细胞的相对存活率,确定BPA最适染毒剂量;最后先用菟丝子黄酮和TM3细胞共同培养4 h后,再用BPA干扰TM3细胞24 h。通过ELISA检测细胞上清液睾酮(T)和黄体生成素(LH)的含量,并分析菟丝子黄酮对BPA干扰小鼠睾丸间质细胞激素分泌的缓解作用。结果表明,250μg/mL～400μg/mL的菟丝子黄酮(等差法)对TM3细胞作用24 h后,有极显著的增殖效果(P<0.01);80μmol/mL的BPA对TM3细胞作用24 h后,细胞存活率为81%,可作为最适染毒剂量;菟丝子黄酮保护4 h后,再用BPA干扰12 h、24 h后,不同浓度的菟丝子黄酮可显著升高TM3细胞睾酮和黄体生成素的水平(P<0.05)。说明菟丝子黄酮对BPA致小鼠睾丸间质细胞睾酮和黄体生成素的分泌异常具有明显的保护作用。

To study the alleviation effect of Cuscuta chinensis flavonoids on BPA-interfering mouse Leydig cell hormone secretion,TM3 mous Leyding cells were used as the research object.Using 1 to 500 μg/mL of Cuscuta chinensis flavonoids interferied cells 24 h,the cell proliferation is detected by MTT method to determine the safe dose;Using 1-200 μmol/mL of bisphenol A (BPA) interfered cells 24 h,to detect cell proliferation by MTT,by the relative survival rate of the TM3 cells,to determine the dose;After the protection of 4 h of traditional Chinese medicine,BPA interfered with 24 h.Using the ELISA method to detect the contents of the testosterone and Luteinizing hormone and secretion.The results showed that 250-400 μg/mL of Cuscuta chinensis flavonoids (equal difference method) had a significant proliferation effect on TM3 cells for 24 h ( P <0.01);The cell survival rate was 81% after 24 h of BPA in 80 μmol/mL.After the protection of 4 h of traditional Chinese medicine,BPA interfered with 12 h and 24 h,and the different concentrations of Cuscuta chinensis flavonoids increased the testosterone level of TM3 cells ( P <0.05).It was showed that Cuscuta chinensis flavonoids has a significant effect on the secretion of testosterone and Luteinizing hormone in mice induced by BPA.