School of Public Health, Beihua University, Jilin, 132013, China.
The First Clinical Medical College of Nanchang University, Nanchang, 330006, China.
Biol Res. 2019 Aug 6;52(1):41. doi: 10.1186/s40659-019-0248-1.
Di-N-butyl-phthalate (DBP) is an endocrine disrupting substance. We investigated the adverse effect of DBP on testis of male rat and reveal its potential mechanism of MAPK signaling pathway involved this effect in vivo and in vitro. Gonadal hormone, sperm quality, morphological change and the activation status of JNK, ERK1/2 and p38 was determined in vivo. Primary Sertoli cell was established and cultivated with JNK, ERK1/2 inhibitors, then determine the cell viability, apoptosis and the expression of p-JNK, p-ERK1/2. Data in this study were presented as mean ± SD and determined by one-way analysis of variance (ANOVA) followed by Bonferroni's test. Difference was considered statistically significant at P < 0.05.
In vivo experiment, DBP impaired the normal structure of testicular tissue, reduced testosterone levels in blood serum, decreased sperm count and increased sperm abnormality, p-ERK1/2 and p-JNK in rat testicular tissue increased in a dose-dependent manner. In vitro studies, DBP could decrease the viability of Sertoli cells and increase p-ERK1/2 and p-JNK. Cell apoptosis in SP600125 + DBP group was significantly lower than in DBP group (P < 0.05). p-JNK was not significantly decreased in SP600125 + DBP group, while p-ERK1/2 was significantly decreased in U0126 + DBP group.
These results suggest that DBP can lead to testicular damage and the activation of ERK1/2 and JNK pathways, the JNK signaling pathway may be primarily associated with its effect.
邻苯二甲酸二正丁酯（DBP）是一种内分泌干扰物质。我们研究了 DBP 对雄性大鼠睾丸的不良影响，并在体内和体外揭示了其涉及 MAPK 信号通路的潜在机制。在体内测定了性腺激素、精子质量、形态变化以及 JNK、ERK1/2 和 p38 的激活状态。建立并培养了原代支持细胞，并用 JNK、ERK1/2 抑制剂进行处理，然后测定细胞活力、细胞凋亡以及 p-JNK、p-ERK1/2 的表达。本研究中的数据以平均值±标准差表示，并通过单向方差分析（ANOVA）后进行 Bonferroni 检验进行确定。P 值<0.05 时认为差异具有统计学意义。
体内实验中，DBP 损害了睾丸组织的正常结构，降低了血清中的睾酮水平，减少了精子计数，增加了精子畸形，大鼠睾丸组织中的 p-ERK1/2 和 p-JNK 呈剂量依赖性增加。体外研究中，DBP 可降低支持细胞的活力并增加 p-ERK1/2 和 p-JNK。SP600125+DBP 组的细胞凋亡明显低于 DBP 组（P<0.05）。SP600125+DBP 组的 p-JNK 没有明显降低，而 U0126+DBP 组的 p-ERK1/2 明显降低。
这些结果表明，DBP 可导致睾丸损伤和 ERK1/2 和 JNK 通路的激活，JNK 信号通路可能与其作用主要相关。
