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目的研究微囊藻毒素-LR(MC-LR)染毒对大鼠睾丸支持细胞凋亡形态以及半胱氨酸天冬氨酸蛋白酶-3(caspase-3)活力的改变。方法自健康18～20日龄SPF级SD雄性大鼠体内分离、培养睾丸支持细胞。用含MC-LR终浓度分别为0(对照)、1、10μg/ml的培养液培养48 h。采用Hoechst荧光染色法检测睾丸支持细胞的凋亡情况;采用caspase-3底物切除法检测caspase-3的活力。结果随着MC-LR染毒剂量的增高,大鼠睾丸支持细胞凋亡率升高。与对照组比较,仅10μg/ml MC-LR染毒的睾丸支持细胞caspase-3活力升高,差异有统计学意义(P<0.05);而1μg/ml MC-LR染毒睾丸支持细胞caspase-3活力略有降低,但差异无统计学意义。结论 MC-LR能诱导睾丸支持细胞发生凋亡,caspase-3在MC-LR诱导睾丸支持细胞凋亡中起重要调控作用。
Objective To study the changes of apoptotic morphology and the activity of caspase-3 in the testicular sertoli cells of microcystin-LR (MC-LR) -treated rats. Methods SPF SD male rats aged from 18 to 20 days of age were isolated and cultured to establish testicular sertoli cells. The cells were cultured for 48 h in medium containing MC-LR final concentrations of 0 (control), 1, 10 μg / ml, respectively. The apoptosis of testicular sertoli cells was detected by Hoechst staining. The activity of caspase-3 was detected by caspase-3 substrate excision. Results With the increase of MC-LR dose, the apoptosis rate of rat testicular sertoli cells increased. Compared with the control group, the viability of caspase-3 in testis supporting cells exposed to 10μg / ml MC-LR increased significantly (P <0.05), while the effect of 1μg / ml MC-LR on the expression of caspase- 3 vitality decreased slightly, but the difference was not statistically significant. Conclusion MC-LR can induce apoptosis of testicular sertoli cells and caspase-3 plays an important role in MC-LR-induced apoptosis of testicular sertoli cells.