中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2010年
5期
461-467
,共7页
黄莉%黄韧%刘寒英%丘剑峰%冯媛瑜%张钰
黃莉%黃韌%劉寒英%丘劍峰%馮媛瑜%張鈺
황리%황인%류한영%구검봉%풍원유%장옥
二噁英%受体蛋白%免疫组织化学%组织分布%生殖毒性
二噁英%受體蛋白%免疫組織化學%組織分佈%生殖毒性
이오영%수체단백%면역조직화학%조직분포%생식독성
Dioxin%Receptor protein%Immunohistochemistry%Tissue distribution%Reproductive toxicity
目的 探讨引起小鼠早期妊娠毒性敏感与相关受体、代谢酶水平的关系,了解子宫组织的敏感性是否与二噁英(TCDD)代谢活化而导致局部毒性蓄积有关.方法 收集TCDD妊娠早期毒性模型小鼠的子宫、肝脏和肾脏组织,用免疫组织化学法对芳香碳氢化合物受体(AhR),受体核转位蛋白(ARNT)和细胞色素氧化酶(Cyp1a2)在组织细胞中分布进行检测.结果 正常妊娠小鼠的子宫、肝脏和肾脏组织细胞质有AhR、Cyp1a2弱阳性信号,细胞核内ARNT有强阳性信号.用不同浓度TCDD处理小鼠,各组织细胞质内的AhR阳性信号随浓度的提高而增加.50 ng/L剂量TCDD暴露使肝脏(18.038±3.0916,P＜0.01),子宫(8.8140±1.4574,P＜0.01)的AhR阳性面积比提高,以及吸光度(A)值显著升高(0.1143±0.0066,0.2399±0.0054,P＜0.01);而肾脏中2种指标均未见显著性差异.相同处理组Cyp1a2变化则主要表现在吸光度的变化,肝脏(0.2047±0.0150,0.31112±0.0107,P＜0.01),子宫(0.1892±0.0232,0.3152±0.0432,P＜0.05)和肾脏(0.2545±0.0202,0.3334±0.0168,P＜0.05)A值均显著升高;组织细胞核内ARNT阳性吸光度在肝脏(0.7496±0.0469,0.2458±0.006,P＜0.01)和子宫(0.4766±0.0359,0.2139±0.0253,P＜0.01)显著下降,在肾脏则表现为阳性信号从细胞核进入细胞质.100 ng/L剂量TCDD暴露则引起所有组织AhR和Cyp1a2阳性显著增强;肝脏和子宫ARNT信号消失,肾脏中ARNT信号从核内转入细胞质.结论 肝脏、子宫和肾脏AhR和CYP1A2酶可受TCDD类物质诱导,细胞信号面积随TCDD暴露量的增加而增大;肝脏和子宫细胞核内的ARNT随TCDD暴露量的增加,细胞信号减弱至消失,但肾脏ARNT进入胞质.说明子宫与肝脏具有相同的TCDD诱导特征,具有活化内外源性有毒化合物的功能,这应该是引起小鼠早期生殖毒性的原因之一.
目的 探討引起小鼠早期妊娠毒性敏感與相關受體、代謝酶水平的關繫,瞭解子宮組織的敏感性是否與二噁英(TCDD)代謝活化而導緻跼部毒性蓄積有關.方法 收集TCDD妊娠早期毒性模型小鼠的子宮、肝髒和腎髒組織,用免疫組織化學法對芳香碳氫化閤物受體(AhR),受體覈轉位蛋白(ARNT)和細胞色素氧化酶(Cyp1a2)在組織細胞中分佈進行檢測.結果 正常妊娠小鼠的子宮、肝髒和腎髒組織細胞質有AhR、Cyp1a2弱暘性信號,細胞覈內ARNT有彊暘性信號.用不同濃度TCDD處理小鼠,各組織細胞質內的AhR暘性信號隨濃度的提高而增加.50 ng/L劑量TCDD暴露使肝髒(18.038±3.0916,P＜0.01),子宮(8.8140±1.4574,P＜0.01)的AhR暘性麵積比提高,以及吸光度(A)值顯著升高(0.1143±0.0066,0.2399±0.0054,P＜0.01);而腎髒中2種指標均未見顯著性差異.相同處理組Cyp1a2變化則主要錶現在吸光度的變化,肝髒(0.2047±0.0150,0.31112±0.0107,P＜0.01),子宮(0.1892±0.0232,0.3152±0.0432,P＜0.05)和腎髒(0.2545±0.0202,0.3334±0.0168,P＜0.05)A值均顯著升高;組織細胞覈內ARNT暘性吸光度在肝髒(0.7496±0.0469,0.2458±0.006,P＜0.01)和子宮(0.4766±0.0359,0.2139±0.0253,P＜0.01)顯著下降,在腎髒則錶現為暘性信號從細胞覈進入細胞質.100 ng/L劑量TCDD暴露則引起所有組織AhR和Cyp1a2暘性顯著增彊;肝髒和子宮ARNT信號消失,腎髒中ARNT信號從覈內轉入細胞質.結論 肝髒、子宮和腎髒AhR和CYP1A2酶可受TCDD類物質誘導,細胞信號麵積隨TCDD暴露量的增加而增大;肝髒和子宮細胞覈內的ARNT隨TCDD暴露量的增加,細胞信號減弱至消失,但腎髒ARNT進入胞質.說明子宮與肝髒具有相同的TCDD誘導特徵,具有活化內外源性有毒化閤物的功能,這應該是引起小鼠早期生殖毒性的原因之一.
목적 탐토인기소서조기임신독성민감여상관수체、대사매수평적관계,료해자궁조직적민감성시부여이오영(TCDD)대사활화이도치국부독성축적유관.방법 수집TCDD임신조기독성모형소서적자궁、간장화신장조직,용면역조직화학법대방향탄경화합물수체(AhR),수체핵전위단백(ARNT)화세포색소양화매(Cyp1a2)재조직세포중분포진행검측.결과 정상임신소서적자궁、간장화신장조직세포질유AhR、Cyp1a2약양성신호,세포핵내ARNT유강양성신호.용불동농도TCDD처리소서,각조직세포질내적AhR양성신호수농도적제고이증가.50 ng/L제량TCDD폭로사간장(18.038±3.0916,P＜0.01),자궁(8.8140±1.4574,P＜0.01)적AhR양성면적비제고,이급흡광도(A)치현저승고(0.1143±0.0066,0.2399±0.0054,P＜0.01);이신장중2충지표균미견현저성차이.상동처리조Cyp1a2변화칙주요표현재흡광도적변화,간장(0.2047±0.0150,0.31112±0.0107,P＜0.01),자궁(0.1892±0.0232,0.3152±0.0432,P＜0.05)화신장(0.2545±0.0202,0.3334±0.0168,P＜0.05)A치균현저승고;조직세포핵내ARNT양성흡광도재간장(0.7496±0.0469,0.2458±0.006,P＜0.01)화자궁(0.4766±0.0359,0.2139±0.0253,P＜0.01)현저하강,재신장칙표현위양성신호종세포핵진입세포질.100 ng/L제량TCDD폭로칙인기소유조직AhR화Cyp1a2양성현저증강;간장화자궁ARNT신호소실,신장중ARNT신호종핵내전입세포질.결론 간장、자궁화신장AhR화CYP1A2매가수TCDD류물질유도,세포신호면적수TCDD폭로량적증가이증대;간장화자궁세포핵내적ARNT수TCDD폭로량적증가,세포신호감약지소실,단신장ARNT진입포질.설명자궁여간장구유상동적TCDD유도특정,구유활화내외원성유독화합물적공능,저응해시인기소서조기생식독성적원인지일.
Objective To investigate relationships between high sensitivity to toxicity in NIH mice during early pregnancy and level of receptors and metabolizing enzymes to 2,3,7,8-tetrachlorodibenzo-pdioxin (TCDD), and to elucidate whether the sensitivity is associated with the TCDD accumulation induced by metabolic activation in the uterus. Methods Liver, kidney and uterus tissues in mice models of toxicity exposure during the early pregnancy were collected and fixed. Distribution of the aryl hydrocarbon receptor (AhR), aryl hydrocarbon receptor nuclear translocator (ARNT) and cytochrome P4501a2 (Cyp1a2) was then determined by immunohistochemical staining. Results In unexposed mice, the tissues of liver, kidney and uterus, displayed weak-positive signal of AhR and Cyp1a2 in cytoplasma and strong-positive signal of ARNT in the nuclei. The positive signal of cytoplasma AhR in these three types of tissues was enhanced to varying degrees along with increasing dose of TCCD exposure. Exposure to 50 ng/L TCDD increased both the percentage of AhR-positive area in liver (18.038±3.0916,P＜0.01)and uterus (8.8140±1.4574,P＜0.01) and optic density of AhR signals(0.1143±0.0066,0.2399±0.0054, P＜0.01 ), while no change was found in kidney tissues. With the same treatment, changes in Cyp1a2 were mainly manifested by significant increase in optic density of Cypla2 signals in liver (0.31112±0.0107, P＜0.01 ), uterus (0.3152±0.0432, P＜0.05) and kidney (0.3334±0.0168, P＜0.05 ), while the optic density of positive ARNT signals in nuclei decreased significantly in liver (0.2458±0.006, P＜0.01 )and uterus(0.2139±0.0253, P＜0.01 ), was largely unchanged but associated with a translocation from nuclei to cytoplasma in kidney. Exposure to 100 ng/L TCDD resulted in enhanced AhR and Cyp1a2 positive signals in all the tissues examined, abolition of ARNT signals in liver and uterus,and traslocation of ARNT signal from nuclei to cytoplasma in the kidney. Conclusions The AhR and Cyp1a2 in liver, uterus and kidney induced by TCDD-like agents may show increasing percentage of signalpositive area along with higher dose of TCDD exposure. In contrast, ARNT signal is weakened and subsequently abolished in nuclei of liver and uterus cells, and displays a traslocation from nuclei to cytoplasma in the kidney. These findings indicate that liver and uterus may share the same TCDD-induced signaling pathway which activates the exogenous toxic agents and should be one of the factors causing sensitivity to TCDD during early pregnancy in mice.
