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目的利用体外培养的大鼠睾丸支持细胞研究微囊藻毒素-LR(MC-LR)对细胞中凋亡相关蛋白表达水平的影响。方法大鼠睾丸细胞分别染毒0(对照)、0.5、1、10、20μg/ml MC-LR溶液后培养24和48 h,采用MTT法检测细胞活性。调整细胞密度为4×106～5×106/ml,分别染毒含0(对照)、1、10μg/ml MC-LR无血清培养液后培养24和48 h,采用Western blot法检测细胞中凋亡相关蛋白[p53基因调节蛋白(P53)、B细胞淋巴瘤/白血病-2蛋白(Bcl-2)、前凋亡蛋白(Bax)]表达水平的变化。结果与对照组比较,10和20μg/ml MC-LR染毒24和48 h后大鼠睾丸支持细胞活性均降低(P<0.05)。与对照组相比,1、10μg/ml MC-LR染毒24 h以及10μg/ml MC-LR染毒48 h时大鼠睾丸支持细胞中P53蛋白相对表达量均显著升高(P<0.05),10μg/ml MC-LR染毒24、48 h大鼠睾丸支持细胞中Bax蛋白相对表达量均显著升高(P<0.05),Bcl-2相对表达量显著降低(P<0.05)。结论较高剂量MC-LR能明显抑制支持细胞活性,凋亡相关蛋白P53、Bcl-2、Bax参与调控MC-LR诱导的支持细胞凋亡。
OBJECTIVE: To study the effect of microcystin-LR (MC-LR) on the expression of apoptosis-related proteins in rat testicular sertoli cells in vitro. Methods Rat testicular cells were exposed to 0 (control), 0.5, 1, 10, 20μg / ml MC-LR solution for 24 and 48 hours respectively. The cell viability was measured by MTT assay. The cell density was adjusted to 4 × 106 ~ 5 × 106 / ml, and the cells were incubated with 0 (control), 1, 10μg / ml MC-LR serum-free medium for 24 and 48 h, respectively. The expressions of P53, Bcl-2 and Bax were compared between the two groups. Results Compared with the control group, the viability of the testis cells in 10 and 20 μg / ml MC-LR cells after 24 and 48 h exposure were significantly decreased (P <0.05). Compared with the control group, the relative expression of P53 protein in the testicular sertoli cells of 1, 10μg / ml MC-LR for 24 h and 10μg / ml MC-LR for 48 h were significantly increased (P <0.05) (P <0.05), and the relative expression of Bcl-2 was significantly decreased (P <0.05). Conclusions Higher doses of MC-LR can significantly inhibit the activity of supporting cells. The apoptosis-related proteins P53, Bcl-2 and Bax are involved in the regulation of MC-LR-induced apoptosis.