论文部分内容阅读
目的探讨胰岛素样因子3(INSL3)对体外培养的小鼠睾丸引带细胞增殖和收缩活性的影响。方法手术放大镜解剖出3日龄雄性昆明小鼠的睾丸引带组织,进行原代细胞培养后传代。将传代细胞随机分为正常对照组和实验组,实验组加入INSL3,浓度分别为3.3×10-3μmol.L-1、3.3×10-4μmol.L-1、3.3×10-5μmol.L-1及3.3×10-6μmol.L-1,分别持续作用12 h、24 h、48 h,利用细胞计数试剂盒(CCK-8)检测其细胞增殖情况;细胞免疫荧光和流式细胞术检测其细胞纤维状肌动蛋白(F-actin)的结构变化及表达情况。结果不同浓度INSL3作用后的不同时间点,检测细胞增殖和收缩活性的相关指标,结果均存在时间-剂量效应,且差异有统计学意义(P<0.05)。正常对照组及各实验组细胞均呈持续性增殖,24 h后增殖更明显。与正常对照组比较,INSL3可刺激睾丸引带细胞骨架重塑,细胞周边肌动蛋白丝带增多,胞质中微丝F-actin明显粗大、变长,细胞中F-actin表达量增加。各实验组中以3.3×10-3μmol.L-1组变化尤其明显。结论 INSL3对睾丸引带细胞增殖和收缩活性有直接促进作用,可能参与睾丸引带发育甚或睾丸下降过程的调节。
Objective To investigate the effect of insulin-like factor 3 (INSL3) on the proliferation and contractile activity of tape-bearing cells in vitro in mice. Methods Testicular tapes of 3-day-old male Kunming mice were dissected with a surgical magnifying glass and subcultured in primary cells. The passage cells were randomly divided into normal control group and experimental group. INSL3 was added to the experimental group at the concentrations of 3.3 × 10-3μmol.L-1, 3 × 10-4μmol.L-1, 3 × 10-5μmol.L-1 And 3.3 × 10-6μmol.L-1 for 12 h, 24 h, 48 h, respectively. The proliferation of cells was detected by CCK-8. The cells were detected by immunofluorescence and flow cytometry Structural changes and expression of F-actin. Results The indexes of proliferation and contractile activity of INSL3 cells treated with different concentrations of INSL3 for different time points showed the time-dose effect and the difference was statistically significant (P <0.05). The normal control group and the experimental group cells showed a sustained proliferation, proliferation was more obvious after 24 h. Compared with the normal control group, INSL3 could stimulate the remodeling of cytoskeleton of testis, the increase of ribbons of peripheral actin, the actin filaments in the cytoplasm were obviously thicker and longer, and the expression of F-actin in the cells increased. The experimental group in 3.3 × 10-3μmol.L-1 group of changes is particularly evident. Conclusion INSL3 has direct effect on the proliferation and contractile activity of tape cells in testis, which may be involved in the regulation of testicular tape development or testicular descent.