萝卜硫素对多柔比星诱导的大鼠卵巢损伤的保护作用

Protective effect of sulforaphane on doxorubicin-induced ovarian damage in rats

目的探讨萝卜硫素(sulforaphane,SFN)对多柔比星诱导的大鼠卵巢功能损害的保护作用及机制。方法将60只雌性Wistar大鼠随机分为空白组(不做任何处理,15只,A组)及化疗组(尾静脉注射多柔比星30 mg/kg,连续14 d,45只)。化疗组大鼠再随机分为对照组(不再予其他处理,B组)、萝卜硫素低剂量组(尾静脉注射萝卜硫素0.15 mL/20 g,连续45 d,C组)和萝卜硫素高剂量组(尾静脉注射萝卜硫素0.3 mL/20 g,连续45 d,D组),每组15只。每15天测重,苏木精-伊红(HE)染色观察卵巢组织结构和各级卵泡计数;检测促卵泡激素(folliclestimulatinghormone,FSH)和雌二醇(estradiol 2,E2)水平;Western blot检测Miga1蛋白表达水平;q RT-PCR法检测Miga1-mRNA表达水平。结果 B组大鼠体质量明显低于A组,C、D组则明显高于B组(P<0.05)。B组大鼠动情周期紊乱,C、D组大鼠动情周期部分恢复规律。C、D组血清E2水平显著高于B组,FSH水平显著低于B组。与B组比较,C、D组卵泡形态、结构规则,炎性细胞较少,始基卵泡、初级卵泡、次级卵泡和窦状卵泡数量增加(P<0.05)。B组Miga1蛋白水平低于A组,C、D组的Miga1蛋白水平高于B组(P<0.05),C组显著低于A、D组(P<0.05)。B组Miga1-mRNA水平低于A组,C、D组的Miga1-mRNA水平高于B组(P<0.05),C组显著低于A、D组,差异有统计学意义(P<0.05)。结论萝卜硫素可以保护多柔比星对大鼠卵巢功能的损害,其作用机制可能与上调Miga1相关。

Objective To investigate the protective effect of sulforaphane(SFN) on ovarian damage caused by doxorubicin in rats and its mechanism. Methods A total of 60 female Wistar rats were randomly divided into blank control group(15 female Wistar rats, group A, without any treatment) and chemotherapy group(45 female Wistar rats, 30 mg/kg, 14 days). Female Wistar rats in chemotherapy group were then randomly divided into the control group(no other treatment, group B), the low-dose sulforaphane group(0.15 mL/20 g, 45 days) and the high-dose sulforaphane group(0.3 mL/20 g, 45 days), with 15 animals in each group. Weight was measured every 15 days. The morphology of ovarian tissue and follicular count were observed by hematoxylin-eosin(HE) staining. The levels of FSH and E2 were detected by ELISA. Western blot was used to detect the expression level of Miga1 protein. The expression level of Miga1 mRNA was detected by qRT-PCR. Results The body weight of rats in group B was significantly lower than that in group A, while that in group C and D was significantly higher than group B(P<0.05). The estrous cycle of rats in group B was disturbed, while that of rats in group C and D was partially restored. The level of serum E2 in group C and D were significantly higher than that in group B, and the level of serum FSH was significantly lower than that in group B(P<0.05). Compared with group B, the follicles in group C and D were regular in morphology and structure, with fewer inflammatory cells, and the number of primary follicles, primary follicles, secondary follicles and sinus-like follicles increased(P<0.05). The level of Miga1 protein in group B was lower than that in group A, and that in group C and D was higher than that in group B(P<0.05). The level of Miga1 protein in group C was lower than that in group A, D(P<0.05). The level of Miga1 mRNA in group B was lower than that in group A, and that in group C and D was higher than that in group B(P<0.05). The level of Miga1 mRNA in group C was lower than that in group A, D(P<0.05). Conclusion Sulforaphan can protect the ovarian function of rats induced by doxorubens, its mechanism may be related to the up-regulation of Miga1.