中国比较医学杂志
中國比較醫學雜誌
중국비교의학잡지
CHINESE JOURNAL OF COMPARATIVE MEDICINE
2014年
6期
1-6
,共6页
李军延%乔佩环%张林媛%刘帅%于淼%常兵
李軍延%喬珮環%張林媛%劉帥%于淼%常兵
리군연%교패배%장림원%류수%우묘%상병
己烯雌酚%间质细胞%氧化损伤%睾酮合成
己烯雌酚%間質細胞%氧化損傷%睪酮閤成
기희자분%간질세포%양화손상%고동합성
Diethylstilbestrol%Testis%Rats%Leydig cells%Testosterone%Oxidative damage%Steroidogenesis
目的:己烯雌酚(diethylstilbestrol,DES)可以导致睾丸氧化损伤,而氧化损伤则可能是导致类固醇合成障碍的作用机制之一。本文探讨DES导致睾丸氧化损伤与睾酮合成途径的关系及可能的机制。方法24只健康4周龄雄性Wistar大鼠,随机分为4组,即对照组(玉米油)和DES染毒组(0.1、1.0、10.0μg/kg ),每组6只,皮下注射每日1次,连续染毒8周。染毒结束后称量动物体重及雄性生殖器官和附属生殖器官(睾丸、附睾、前列腺)的重量,用生化方法检测睾丸匀浆中丙二醛( MDA)和活性氧自由基( ROS)的含量,抗氧化酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)的活性变化以及类固醇合成酶3β羟类固醇脱氢酶1(3β-HSD1)、17β羟类固醇脱氢酶3(17β-HSD3)的活性,用放射免疫法测定外周血中睾酮、促黄体生成素(LH)的水平,用RT-PCR检测固醇激素急性调节蛋白(StAR)、P450胆固醇侧链裂解酶(P450scc)、3β羟类固醇脱氢酶1(3β-HSD1)、17β羟类固醇脱氢酶3(17β-HSD3) mRNA水平的表达变化。结果10.0μg/kg组睾丸、前列腺重量以及脏器系数明显减少,MDA和ROS的氧化程度明显升高,SOD、CAT、GPx的活性降低,3β-HSD1、17β-HSD3的活性分别降低;血清睾酮降低,1.0μg/kg组LH降低,整个染毒区间呈现剂量效应关系;10.0μg/kg组StAR、P450scc、3β-HSD1、17β-HSD3 mRNA表达减少,1.0μg/kg组StAR、3β-HSD1依然减少。结论 DES暴露干扰了大鼠睾丸氧化/抗氧化平衡,同时导致包括睾酮水平降低在内的雄性生殖危害。 DES导致GPx、CAT酶活力降低,抑制睾丸类固醇合成途径中P450scc,3β-HSD1 mRNA的表达水平,以及3β-HSD1活性降低导致睾酮减少。推测该途径可能是DES干扰类固醇合成的毒性作用机制之一。
目的:己烯雌酚(diethylstilbestrol,DES)可以導緻睪汍氧化損傷,而氧化損傷則可能是導緻類固醇閤成障礙的作用機製之一。本文探討DES導緻睪汍氧化損傷與睪酮閤成途徑的關繫及可能的機製。方法24隻健康4週齡雄性Wistar大鼠,隨機分為4組,即對照組(玉米油)和DES染毒組(0.1、1.0、10.0μg/kg ),每組6隻,皮下註射每日1次,連續染毒8週。染毒結束後稱量動物體重及雄性生殖器官和附屬生殖器官(睪汍、附睪、前列腺)的重量,用生化方法檢測睪汍勻漿中丙二醛( MDA)和活性氧自由基( ROS)的含量,抗氧化酶超氧化物歧化酶(SOD)、過氧化氫酶(CAT)、穀胱甘肽過氧化物酶(GPx)的活性變化以及類固醇閤成酶3β羥類固醇脫氫酶1(3β-HSD1)、17β羥類固醇脫氫酶3(17β-HSD3)的活性,用放射免疫法測定外週血中睪酮、促黃體生成素(LH)的水平,用RT-PCR檢測固醇激素急性調節蛋白(StAR)、P450膽固醇側鏈裂解酶(P450scc)、3β羥類固醇脫氫酶1(3β-HSD1)、17β羥類固醇脫氫酶3(17β-HSD3) mRNA水平的錶達變化。結果10.0μg/kg組睪汍、前列腺重量以及髒器繫數明顯減少,MDA和ROS的氧化程度明顯升高,SOD、CAT、GPx的活性降低,3β-HSD1、17β-HSD3的活性分彆降低;血清睪酮降低,1.0μg/kg組LH降低,整箇染毒區間呈現劑量效應關繫;10.0μg/kg組StAR、P450scc、3β-HSD1、17β-HSD3 mRNA錶達減少,1.0μg/kg組StAR、3β-HSD1依然減少。結論 DES暴露榦擾瞭大鼠睪汍氧化/抗氧化平衡,同時導緻包括睪酮水平降低在內的雄性生殖危害。 DES導緻GPx、CAT酶活力降低,抑製睪汍類固醇閤成途徑中P450scc,3β-HSD1 mRNA的錶達水平,以及3β-HSD1活性降低導緻睪酮減少。推測該途徑可能是DES榦擾類固醇閤成的毒性作用機製之一。
목적:기희자분(diethylstilbestrol,DES)가이도치고환양화손상,이양화손상칙가능시도치류고순합성장애적작용궤제지일。본문탐토DES도치고환양화손상여고동합성도경적관계급가능적궤제。방법24지건강4주령웅성Wistar대서,수궤분위4조,즉대조조(옥미유)화DES염독조(0.1、1.0、10.0μg/kg ),매조6지,피하주사매일1차,련속염독8주。염독결속후칭량동물체중급웅성생식기관화부속생식기관(고환、부고、전렬선)적중량,용생화방법검측고환균장중병이철( MDA)화활성양자유기( ROS)적함량,항양화매초양화물기화매(SOD)、과양화경매(CAT)、곡광감태과양화물매(GPx)적활성변화이급류고순합성매3β간류고순탈경매1(3β-HSD1)、17β간류고순탈경매3(17β-HSD3)적활성,용방사면역법측정외주혈중고동、촉황체생성소(LH)적수평,용RT-PCR검측고순격소급성조절단백(StAR)、P450담고순측련렬해매(P450scc)、3β간류고순탈경매1(3β-HSD1)、17β간류고순탈경매3(17β-HSD3) mRNA수평적표체변화。결과10.0μg/kg조고환、전렬선중량이급장기계수명현감소,MDA화ROS적양화정도명현승고,SOD、CAT、GPx적활성강저,3β-HSD1、17β-HSD3적활성분별강저;혈청고동강저,1.0μg/kg조LH강저,정개염독구간정현제량효응관계;10.0μg/kg조StAR、P450scc、3β-HSD1、17β-HSD3 mRNA표체감소,1.0μg/kg조StAR、3β-HSD1의연감소。결론 DES폭로간우료대서고환양화/항양화평형,동시도치포괄고동수평강저재내적웅성생식위해。 DES도치GPx、CAT매활력강저,억제고환류고순합성도경중P450scc,3β-HSD1 mRNA적표체수평,이급3β-HSD1활성강저도치고동감소。추측해도경가능시DES간우류고순합성적독성작용궤제지일。
Objective It is well known that diethylstilbestrol ( DES ) can result in testicular oxidative injury , and one of its mechanisms of action is leading to dysfunction of steroidogenesis .The aim of this study was to investigate the relationship between testicular oxidative injury caused by DES and the key synthetase activities for the synthesis pathway of steroidogenesis and the possible mechanism .Methods Twenty-four 4-wk-old male Wistar albino rats were randomly divided into 4 groups , 6 rats each.Three doses of DES (0.1, 1.0 and 10 μg/kg· d) groups and a vehicle (corn oil) control group , were respectively administered by subcutaneous injection once a day for eight weeks .The rats were sacrificed after 8 weeks treatment and the body weight , testis, epididymis, prostate were weighed, respectively.The testicular tissues were homogenized and the oxidation of MDA and ROS , the activity changes of antioxidases SOD, CAT and GPx, as well asthe activities of steroid synthetases 3β-HSD1 and 17β-HSD3 were determined by biochemical measurement.The levels oftestosterone and LH in peripheral blood were measured by radioimmunoassay .The intensities of expression of StAR,P450scc, 3β-HSD1, 17β-HSD3-mRNA were detected by PCR.Results In the 10.0 μg/kg dose group, the weights andorgan coefficients of testis and prostate were decreased significantly , the oxidation of MDA and ROS was increased distinctlyand the activities of SOD, CAT, GPx, 3β-HSD1 and 17β-HSD3 were reduced.The concentration of serum testosterone wasdecreased in the 10.0 μg/kg dose group.In the 10.0 μg/kg and 1.0 μg/kg dose groups, the decline of LH levelpresented a dose-dependent manner, and the intensities of immunochemical positive staining for StAR , P450scc, 3β-HSD1and 17β-HSD3 mRNA were decreased.Conclusions DES exposure results in disturbance of the oxidant /antioxidantbalance and decline of testosterone level that induces reproductive impairment in male rats .DES induces reductions of bothGPx and 3β-HSD activities which cause the decrease of testosterone synthesis .The expression of P450scc and 3β-HSDmRNA,which are the key synthetases in biosynthetic pathway of steroidogenesis , are inhibited by DES, and it isspeculated that the disturbance of steroidogenic synthesis enzymes may be one of the mechanisms of toxic effects of DES .