氨磷汀和维生素E对顺铂导致大鼠卵巢功能损伤作用机制探讨

Role of amifostine and vitamin E on ovary function damage caused by cisplatin in rats

目的 探讨氨磷汀和维生素E是否可减轻顺铂对大鼠卵巢功能的损伤及其作用机制。方法 将40只雌性SD大鼠随机分为顺铂组(A组)、顺铂+维生素E组(B组)、顺铂+氨磷汀组(C组)、空白对照组(D组)4组,每组10只。D组大鼠正常饲养。A、B、C组大鼠于实验的第1天、第8天分别给予顺铂6 mg/kg腹腔注射;B组大鼠再于实验开始后每日给予维生素E 100 mg/kg灌胃,共13 d;C组大鼠再于实验第1天、第8天分别给予氨磷汀200 mg/kg腹腔注射。实验第14天处死大鼠,取腹主动脉血清,ELISA法检测雌二醇(E2)和卵泡刺激素(FSH)值。取左卵巢组织4%甲醛固定石蜡包埋切片后HE染色,光镜下对各组大鼠卵巢的各级卵泡记数;取右卵巢组织提取细胞胞质,通过WST-1法测胞质超氧化物歧化酶(SOD)活性值。结果 A组原始卵泡数为(12.70±2.79)个,与B组的(19.30±2.79)个比较,差异有统计学意义,P=0.001;与C组的(22.00±6.16)个比较,差异有统计学意义,P<0.001。A组生长卵泡数为(7.90±0.32)个,与B组的(12.70±4.19)个、C组的(17.70±3.34)个比较,差异均有统计学意义,均P<0.05;B组与C组比较,差异亦有统计学意义,P=0.020。A组血清E2为(21.94±2.06) ng/L,与C组的(35.25±3.88) ng/L比较,差异有统计学意义,P<0.001;B组的为(23.70±2.05) ng/L,与C组比较,差异亦有统计学意义,P<0.001。C组血清FSH为(31.08±3.36) ng/mL,与A组的(56.87±8.76) ng/mL、B组的(52.48±8.12) ng/mL比较,差异均有统计学意义,均P<0.001。A组卵巢组织胞质SOD活性值为(129.59±9.42) u/mgprot,与B组的(242.25±15.71) u/mgprot、C组的(246.74±24.03) u/mgprot比较,差异有统计学意义,均P<0.001。结论 顺铂可对大鼠造成卵巢激素生成功能和组织结构损伤。氨磷汀和维生素E在一定程度上可减轻这种损伤,对卵巢起保护作用,其机制可能与两者减轻顺铂诱导产生的氧自由基损伤有关。

Objective To investigate whether amifostine and vitamin E given concomitantly with cisplatin is effective in preventing cisplatin-induced ovarian damage and the potential mechanism in rats.Methods A total of 40female SpragueDawley rats were randomized into four Groups:cisplatin alone group(Group A),cisplatin+vitamin E group(Group B),cisplatin+amifostine group(Group C)and control group(Group D),10each.Rats in the Group D were raised normally without any treatment.Rats in Group A,Group B and Group C were injected intraperitoneally with cisplatin 6mg/kg on the first day and 8th day;rats in Group B were given vitamin E 100mg/kg via gavage administration daily and the administration lasted for 13days.Rats in Group C were injected intraperitoneally with amifotine 200mg/kg on the first day and8th day.Rats were executed on the 14th day of the experiment,abdominal artery serum was collected,and estradiol(E_2)and follicle-stimulating hormone(FSH)were detected by ELISA.The left ovary has been fixed in formaldehyde,embedded in paraffin wax,sectioned and stained with HE staining,follicles of different developmental stages in ovarian tissue were counted under light microscope.The contents of superoxide dismutase(SOD)activity in cytoplasm of right ovarian tissues were detected using WST-1assay.Results The number of primordial follicles:Group A (12.70±2.79)was significantly decreased compared with Group B(19.30±2.79),and the difference was statistically significant(P=0.001);Group A was significantly decreased compared with Group C (22.00±6.16),and the difference was statistically significant(P<0.001).The number of developing follicles:Group A (7.90±0.32)was significantly decreased compared with Group B(12.70±4.19),and the difference was statistically significant(P<0.001);Group A was significantly decreased compared with and Group C(17.70±3.34),and the difference was statistically significant(P0.05).Compared with Group B,the number of developing follicles was decreased in the Group C,and the difference was statistically significant(P=0.020).The content of estradiol in serum:Group A (21.94±2.06)ng/L was significantly decreased compared with Group C(35.25±3.88)ng/L,and the difference was statistically significant(P<0.001);Group B(23.70±2.05)ng/L was significantly decreased compared with Group C,and the difference was statistically significant(P<0.001);The content FSH in serum:Group A (56.87±8.76)ng/ml was increased compared with Group C(31.08±3.36)ng/ml,and the difference was statistically significant(P<0.001);Group B (52.48±8.12)ng/ml was increased compared with Group C,and the difference was statistically significant(P<0.001).The activity of SOD in the cytoplasm of ovarian tissue in Group A was(129.59±9.42)u/mgprot,which was significantly different from that in Group B(242.25±15.71)u/mgprot and Group C (246.74±24.03)u/mgprot,P<0.001.Conclusions Cisplatin can cause damage to the ovarian tissue structure and hormone function in rats,while amifostine and vitamin E can alleviate the damage and protect the ovary to a certain extent.The mechanism may be related to the reduction of oxygen free radical damage by amifostine and vitamin E induced by cisplatin.