他克林对脂多糖诱导小鼠睾丸支持细胞炎性损伤的保护作用.

[Objective] To analyze the protective effect of tacrine on lipopolysaccharide (LPS)-induced inflammatory damage in mouse testicular sertoli cells (TM4), and to further explore the function of tacrine from an inflammatory perspective, in order to provide more drug options for the use of anti-inflammatory drugs related to reproductive diseases and sperm quality enhancement in male animals during animal production. [Method] The CCK8 method was used to detect the appropriate damage conditions for TM4 induced by 0. 01, 0. 10, l. 00, 10. 00 and 100. 00 µ.g/mL LPS for 3. 0, 6. 0, 12. 0 and 24. 0 hours, respectively; And to analyze the appropriate protection conditions for TM4 pretreated by 0. 01,0. 10, 1. 00, 10. 00, 100. 00, 1000. 00,10 000. 00 and 100 000. 00 µ.g/mL tacrine for 0. 5, 1. 0 and 2. 0 hours prior to the addition of LPS or treated for 0. 5, 1. 5, 3. 0, 6. 0, 12. 0 and 24. 0 hours after the addition of LPS. The mRNA expression levels of TNF-α and IL-6 genes were measured by fluorescent quantitative PCR(qRT-PCR). The expression levels of TNF-α and IL-6 proteins in cell supernatants were further evaluated by ELISA, and the expression of NF - KB protein were analyzed by immunoprotein blotting (WB). [Result] There were differences in the effects of LPS on the proliferation of TM4 under different concentrations and different treatment times, and the inflammatory damage induced by 1. 00 µ.g/mL LPS treatment of TM4 for 12 hours was significantly lower than that in the blank group (P < 0. 01, the same as below). The protective effect of tacrine on TM4 was time and dose-dependent, with the TM4 proliferation rate of 10 000. 00 µ.g/mL tacrine pretreatment for 2. 0 hours being significantly (P < 0. 05, the same as below) lower than that of the blank group, and 100 000. 00 µ.g/ mL tacrine pretreatment for 0. 5 and 1. 0 hours or post-treatment for 0. 5, 1. 5,3. 0,6. 0, 12. 0 and the TM4 proliferation rate at 24. 0 hours was highly significantly lower than that of the blank group, while the best protection was achieved by 0. 01 µ.g/mL tacrine post-treatment for 6. 0 hours, which had a significantly higher TM4 proliferation rate than that of the blank group. Compared with the blank group, LPS significantly up-regulated the mRNA and protein expression of TM4 IL-6 and TNF-α genes, while 0. 01 µ.g/mL tacrine significantly down-regulated their expression compared with the LPS group. LPS significantly up-regulated NF-KB protein expression in TM4 compared to the blank group; 1. 00, 100. 00 and 10 000. 00 µ.g/mL tacrine significantly up-regulated NF-KB protein expression, while 0. 01 µ.g/mL tacrine decreased NF-KB protein expression compared to the LPS group but the difference was not significant (P >0. 05). [Conclusion] 0. 01 µ.g/mL tacrine post-treatment for 6. 0 hours showed the best protective effect on TM4, and the mechanism may be related to the down-regulation of mRNA and protein expression of IL-6 and TNF-α genes. [ABSTRACT FROM AUTHOR]