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目的通过观察含雷公藤多苷(GTW)、菟丝子黄酮药物血清对体外培养的大鼠睾丸生精细胞增殖、细胞周期及凋亡的影响,探讨菟丝子黄酮干预GTW所致生殖损伤的作用机制。方法体外培养大鼠睾丸生精细胞,将其分为空白组、9mg GTW组(多苷1号组)、9mg GTW+菟丝子黄酮组(黄酮1号组)、12mg GTW组(多苷2号组)、12mg GTW+菟丝子黄酮组(黄酮2号组),加不同含药血清,于培养24h、48h后,通过MTT法检测含药血清对生精细胞增殖的影响;采用流式细胞仪检测含药血清对生精细胞细胞周期及凋亡的影响。结果生精细胞增殖方面:血清作用24h、48h时,黄酮1号组OD值明显高于多苷1号组;黄酮2号组OD值高于多苷2号组;均具有明显统计学差异(P<0.05)。生精细胞S期细胞百分比:黄酮2号组较多苷2号组明显降低(P<0.05);而黄酮1号组与多苷1号组比较无统计学意义(P>0.05)。生精细胞凋亡方面:黄酮1号组较多苷1号组明显降低(P<0.05);而黄酮2号组与多苷2号组比较无统计学意义(P>0.05)。结论GTW可使体外培养幼鼠生精细胞增殖能力下降,细胞凋亡增多。菟丝子黄酮可干预生精细胞的分裂周期,同时对生精细胞凋亡有明显的调控作用。
OBJECTIVE: To investigate the effects of GTW and Dodder seed flavonoids on the proliferation, apoptosis and cell cycle of spermatogenic cells in vitro in rats, and to explore the mechanism of the effect of cuscuta flavone on GTW-induced reproductive injury. Methods Spermatogenic cells were cultured in vitro and divided into blank group, 9mg GTW group (polyglycosidase 1 group), 9mg GTW + diceros flavanone group (flavonoid 1 group), 12mg GTW group (polyglycoside 2 group) , 12mg GTW + dodder flavonoids (flavonoids No.2 group), different serums were added, and the effect of containing serum on the proliferation of spermatogenic cells was detected by MTT assay 24h, 48h after culture. The serum containing serum was detected by flow cytometry Effect on Cell Cycle and Apoptosis of Spermatogenic Cells. Results In the aspect of proliferation of spermatogenic cells, the OD value of flavonoid 1 group was significantly higher than that of polyglucoside 1 group when serum was applied for 24 and 48 hours. The OD value of flavonoid 2 group was higher than that of polyglycoside 2 group, with significant difference P <0.05). The percentage of S phase cells in spermatogenic cells was significantly lower in flavonoid 2 than that in group 2 (P <0.05), while there was no significant difference in flavonoid 1 and polyglycoside 1 (P> 0.05). In the aspect of apoptosis of spermatogenic cells, the flavone 1 group was significantly lower than that of the 1 group (P <0.05), while there was no significant difference between the flavonoid 2 group and the glycoside 2 group (P> 0.05). Conclusion GTW can inhibit the proliferation of spermatogenic cells in vitro and increase the apoptosis rate of germ cells in vitro. Dodder flavonoids can interfere with the spermatogenic cell division cycle, while apoptosis of spermatogenic cells have obvious regulatory role.