没食子酸拮抗环磷酰胺损伤睾丸支持细胞波形蛋白的研究

Gallic acid antagonizes the distribution and expression of Vimentin injury induced by cyclophospha- mide in mice

目的观察没食子酸（GA）拮抗环磷酰胺干扰睾丸支持细胞波形蛋白（Vimentin）分布表达的作用。方法用SK—N．SH细胞株建立3周龄C57BL／6j小鼠移植瘤模型，实验随机对照组：（1）荷瘤鼠磷酸盐缓冲液（PBS）组（PBS组），每只荷瘤鼠腹腔注射PBS0．5ml，阴性对照；（2）荷瘤鼠环磷酰胺组（cP组），每只荷瘤鼠腹腔注射cP75mg／kg，2次／周，连续2周，阳性对照。（3）非荷瘤正常鼠组（Nc组），用于检测小鼠生殖功能对照；实验组：（1）没食子酸组（GA组），每只荷瘤鼠腹腔注射GA250mg／kg，1次／2d，连续2周；（2）荷瘤鼠没食子酸加环磷酰胺组（GA＋CP组），GA在cP给药前24h给药，每只荷瘤鼠给药剂量、方式、时间同GA、CP组，每组10只C57BL／6j小鼠。比较各组睾丸绝对质量及脏器系数，分别应用免疫组织化学、逆转录一聚合酶链反应（RT—PCR）、Westernblot检测睾丸支持细胞Vimentin分布、mRNA和蛋白水平变化。结果CP治疗后睾丸脏器系数与PBS组比较显著降低（P〈0．05），GA干预后升高（P〈0．05）。单用GA治疗后与PBS组差异无统计学意义（P〉0．05）；cP治疗后睾丸支持细胞Vimentin分布紊乱，mRNA及蛋白表达与PBS组比较显著下调（P〈0．01），GA干预后睾丸支持细胞Vimentin分布与PBS组类似，mRNA和蛋白表达较cP组显著上调（P〈0．01），与PBS组比较差异无统计学意义（P〉0．05）。结论GA拈抗环磷酰胺干扰睾丸支持细胞Vimentin正常分布表达作用，可作为环磷酰胺损伤生殖功能的干预药物。

Objective To observe that GA antagonized testicular cytoskeleton protein vimentin injury induced by CP. Methods The 3-week-old immunoeompetcnt C57BL/6j mice were transplanted with SK-N-SH cells,distribution of randomized block design, 10 of each, the control groups： （ 1 ） PBS group, 0. 5ml PBS per bearing tumor mouse,intraperitoneally （i. p） ; （2） CP group,75mg/kg per bearing tumor mouse, i. p, twice a week, continuous two weeks ; （ 3 ） NC group, as the reproductivity of mice. Experimental groups：（1 ）GA group, administered gallic acid 250mg/kg i. p per bearing tumor mouse, every other day, continuous two weeks ; （ 2 ） GA plus CP group, GA and CP independent administration, the dose and time with the above. To compare the absolute weight and the relative organ index of the testes. The expression of vimentin mRNA and protein in testes were detected by immunohistochemistry, reverse transcription-polymer- ase chain reaction （RT-PCR） and Western blotting, respectively. Results The testes organ coefficient were significantly decreased after CP administration （ P 〈 0. 01 ）, and restored after GA intervene （ P 〈 0. 05 ）. Compared with PBS group, there was no significant difference after GA administration alone （ P 〉 0. 05 ）. The cytoskeletal protein vimentin in testes was abnormal distributed after CP administration, howev- er,the distribution were similarly between GA plus CP group and PBS group. The vimentin mRNA and pro- tein expression in testes was significantly lower after CP administration （P 〈 0. 01 ） , but that restored after GA intervene （P 〈 0. 01 ）. There were no significant differences between GA plus CP group and PBS group. Conclusion GA may antagonize the toxicity of CP to the testes and stable the normal distribution and expression of cytoskeletal protein vimentin in mice.