论文部分内容阅读
目的探讨邻苯二甲酸二丁酯(DBP)致大鼠发生尿道下裂的作用和可能机制。方法孕SD大鼠14只随机均分为DBP组和对照组。于妊娠第13~19天,DBP组采用DBP 800mg/kg溶入玉米油灌胃,对照组给予玉米油灌胃。妊娠第19天取出胎鼠,鉴别胎鼠性别,并分为尿道下裂组和对照组。提取两组胎鼠的生殖结节;原代培养生殖结节中的成纤维细胞,Western blot法检测同型雄激素受体A(AR-A)和AR-B以及Ⅰ型胶原蛋白的表达,qPCR检测AR mRNA表达,细胞增殖实验检测细胞增殖能力变化。结果 DBP组雄性仔鼠35只,有尿道下裂鼠12只(34.3%);对照组雄性仔鼠41只,无尿道下裂鼠。对照组的尿道开口于生殖结节顶端,尿道下裂组胎鼠的尿道开口于生殖结节腹侧。与对照组比较,尿道下裂组胎鼠的肛门-生殖结节距离短[(1.57±0.09)mm vs.(2.45±0.16)mm],AR-B相对表达量低(细胞质:0.16±0.07vs.3.46±0.98;细胞核:0.13±0.07vs.2.17±0.58),增殖能力下降,Ⅰ型胶原蛋白相对表达量低(0.22±0.09vs.1.27±0.78)(P<0.05)。结论 DBP可降低胎鼠生殖结节成纤维细胞中AR-B和Ⅰ型胶原蛋白相对表达量,降低成纤维细胞的增殖能力,这可能会影响雄性胎鼠生殖器的发育导致尿道下裂的发生。
Objective To investigate the role and possible mechanism of hypospadias in rats induced by dibutyl phthalate (DBP). Methods 14 pregnant SD rats were randomly divided into DBP group and control group. In the first 13-19 days of gestation, DBP group was administrated with corn oil by DBP 800mg / kg, while the control group was given gavage with corn oil. Fetal rats were taken out on the 19th day of pregnancy to identify the gender of the fetus and divided into hypospadias group and control group. The reproductive nodules were extracted from two groups of fetal rats. The primary cultured reproductive nodules were cultured in vitro. The expression of isobaric androgen receptor A (AR-A), AR-B and collagen Ⅰ was detected by Western blot. QPCR AR mRNA expression was detected, cell proliferation assay to detect changes in cell proliferation. Results Thirty-five male pups in DBP group had 12 (34.3%) of hypospadias; 41 male pups in control group had no urethral hypoderm. In the control group, the urethra was opened at the top of reproductive nodules, and the urethra in the hypospadi group was ventrally opened in the reproductive nodules. Compared with the control group, the anogenital-reproductive nodules in the hypospadias group were shorter than those in the control group [(1.57 ± 0.09) mm vs. (2.45 ± 0.16) mm], and AR-B was relatively low (cytoplasm: 0.16 ± 0.07 vs .3.46 ± 0.98; nucleus: 0.13 ± 0.07vs.2.17 ± 0.58), the proliferation ability decreased, the relative expression of type Ⅰ collagen was low (0.22 ± 0.09vs.1.27 ± 0.78) (P <0.05). Conclusion DBP can reduce the relative expression of AR-B and collagen Ⅰ in fetal rat reproductive nodule fibroblasts and decrease the proliferation of fibroblasts, which may affect the development of genitalia in male fetus leads to the occurrence of hypospadias.