CAJ | 학술논문

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PBS 0.5ml,阴性对照;(2)荷瘤鼠环磷酰胺组(CP组),每只荷瘤鼠腹腔注射CP 75 mg/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GA 250 mg/kg,1次/2 d,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24 h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-聚合酶链反应(RT-PCR)、Western blot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拮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.
목적 관찰몰식자산(GA)길항배린선알간우고환지지세포파형단백(Vimentin)분포표체적작용.방법 용SK-N-SH세포주건립3주령C57BL/6j소서이식류모형,실험수궤대조조:(1)하류서린산염완충액(PBS)조(PBS조),매지하류서복강주사PBS 0.5ml,음성대조;(2)하류서배린선알조(CP조),매지하류서복강주사CP 75 mg/kg,2차／주,련속2주,양성대조.(3)비하류정상서조(NC조),용우검측소서생식공능대조;실험조:(1)몰식자산조(GA조),매지하류서복강주사GA 250 mg/kg,1차/2 d,련속2주;(2)하류서몰식자산가배린선알조(GA+ CP조),GA재CP급약전24 h급약,매지하류서급약제량、방식、시간동GA、CP조,매조10지C57BL/6j소서.비교각조고환절대질량급장기계수,분별응용면역조직화학、역전록-취합매련반응(RT-PCR)、Western blot검측고환지지세포Vimentin분포、mRNA화단백수평변화.결과 CP치료후고환장기계수여PBS조비교현저강저(P＜0.05),GA간예후승고(P＜0.05).단용GA치료후여PBS조차이무통계학의의(P＞0.05);CP치료후고환지지세포Vimentin분포문란,mRNA급단백표체여PBS조비교현저하조(P＜0.01),GA간예후고환지지세포Vimentin분포여PBS조유사,mRNA화단백표체교CP조현저상조(P＜0.01),여PBS조비교차이무통계학의의(P＞0.05).결론 GA길항배린선알간우고환지지세포Vimentin정상분포표체작용,가작위배린선알손상생식공능적간예약물.
Objective To observe that GA antagonized testicular cytoskeleton protein vimentin injury induced by CP.Methods The 3-week-old immunocompetent 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-polymerase 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,however,the distribution were similarly between GA plus CP group and PBS group.The vimentin mRNA and protein 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.