邻苯二甲酸二丁酯和苯并[a]芘联合染毒对大鼠睾丸支持细胞的影响

Combined effect of dibutyl phthalate and benzo(a) pyrene in rat sertoli cells in vitro

目的探讨邻苯二甲酸二丁酯和苯并[a]芘单独或联合染毒对大鼠睾丸支持细胞增殖及乳酸分泌的影响。方法分离纯化大鼠睾丸支持细胞,油红O染色鉴定纯度大于90%后,以0.1、1、10、100、500μg/ml邻苯二甲酸二丁酯(dibutyl phthalate,DBP)和0.01、0.1、1、10、100μg/ml苯并[a]芘[benzo(a)pyrene,BaP]单独或依次联合染毒,用MTT法检测对支持细胞增殖的影响,并测定培养液中的乳酸含量。结果DBP染毒后100μg/ml吸光度值[D(490)]显著高于对照组(P<0.01)。BaP染毒各组的D(490)值与对照组比较,无统计学差异。DBP100μg/ml+BaP10μg/ml联合染毒组D(490)值显著上升(P<0.01),而在500μg/ml DBP+50μg/ml BaP联合染毒组D(490)值下降(P<0.05)。DBP100μg/ml组、100μg/ml DBP+10μg/ml BaP组和500μg/ml DBP+50μg/ml DBP组培养液中的乳酸显著增加(P<0.01),DBP500μg/ml组和BaP50μg/ml组培养液中的乳酸含量也增高(P<0.05)。结论一定剂量DBP可刺激支持细胞增殖并增加乳酸分泌,BaP虽不抑制支持细胞的增殖,但可使乳酸分泌增加。二者联合作用,在一定剂量下以DBP的效应为主,但在高剂量可抑制细胞增殖,呈现一定的协同作用。

Objective To investigate the combined effect of dibutyl phthalate （DBP） and benzo （a）pyrene （BaP） on the proliferation and lactate secretion in rat sertoli cells. Methods Rat testicular sertoli cells were isolated, purified, cultured, and identified with oil red O. The purity of rat sertoli cells was 〉 90%. Then the purified sertoli cells were treated with the toxicants at different dose, i.e. DBP （0.1, 1, 10, 100, 500μg/ml）, BaP （0.01, 0.1, 1, 10, 50 μg/ml） and their combination DBP ＋BaP （0.1 ＋0.01, 1 ＋0.1, 10 ＋ 1, 100＋10, 500 ＋50 μg/ml）. The viability of sertoli cells was determined by MTr method and the lactate concentration in the cultured medium was detected. Results As compared with DMSO control, the absor-bance values in 100 μg/ml DBP and DBP ＋ BaP （ 100 ＋ 10 μg/ml） groups were significantly increased （ P 〈 0. 01 ）, but that in 500 μg/ml DBP group was decreased （P〈0.01） and those of BaP groups were of no statistical difference （P〉0.05）. Lactate concentration in 100, 500 μg/ml DBP groups, 50 μg/ml BaP group and DBP ＋ BaP （ 100 ＋ 10,500 ＋ 50 μg/ml） groups were increased （P 〈0.05）. Conclusion DBP at low dose can promote the proliferation and lactate secretion in secretion in sertoli cells, but BaP can only enhance the lactate secretion. The combination of DBP and BaP, if both at low doses, promotes the proliferation of sertoli cells; if both at high doses, inhibits the proliferation of sertoli cells with high dose. The combined effect of DBP and DaP is synergistic.