CAJ | 학술논문

目的探讨γ射线损伤致不育症小鼠中血睾屏障重要组分Claudin-3、Claudin-11表达的变化. 方法 2011年7月至2011年10月我们选取42只雄性昆明小鼠,6～8周龄,体质量20～25g.随机分为7组,每组6只,分别为空白对照A组；接受60Coγ射线单剂量射线下腹局部照射的B、C、D组,照射剂量分别为2、6、10 Gy;接受6 Gy剂量照射后应用雌二醇1、2、4μg/d干预4周的E、F、G组.于末次给药后2周,处死各组小鼠并收集睾丸,计算各组小鼠睾丸曲细精管分化指数(tubule differentiation index,TDI),应用逆转录-荧光定量聚合酶链式反应检测睾丸组织抑制素βB、Claudin-3、Claudin-11 mRNA的水平,蛋白质印迹法检测睾丸Claudin-11蛋白的表达情况. 结果与A组(100.0)比较,B、C、D组睾丸TDI显著下降(68.5±6.4、35.0±6.1、16.3±5.7),差异均有统计学意义(P＜0.05),且D组睾丸组织抑制素βB mRNA显著降低(0.5±0.2比1.0±0.1),差异有统计学意义(P＜0.05)；C、D组Claudin-3 mRNA水平分别为A组的2.17倍和3.49倍,Claudin-11蛋白水平分别为A组的2.18倍和2.23倍.与C组(35.0±6.1)比较,F、G组TDI显著增高(61.7±7.2、55.8±11.9),差异均有统计学意义(P＜0.05),抑制素βB mRNA虽升高,但差异无统计学意义(均P＞0.05)；F、G组Claudin-3 mRNA水平分别为1.3±0.2和1.6±0.3,低于C组(2.2±0.2),差异均有统计学意义(P＜0.05),且F组Claudin-11 mRNA水平(1.2±0.2)和蛋白表达水平(1.5±0.5)均显著低于C组(1.79±0.2和2.17±0.3),差异均有统计学意义(P＜0.05).TDI与Claudin-3 mRNA、Claudin-11 mRNA水平均呈负相关(rs=-0.884,P＜0.05;rs=-0.758,P＜0.05). 结论 γ射线照射可使小鼠血睾屏障组分Claudin-3、Claudin-11表达水平升高.
목적 탐토γ사선손상치불육증소서중혈고병장중요조분Claudin-3、Claudin-11표체적변화. 방법 2011년7월지2011년10월아문선취42지웅성곤명소서,6～8주령,체질량20～25g.수궤분위7조,매조6지,분별위공백대조A조；접수60Coγ사선단제량사선하복국부조사적B、C、D조,조사제량분별위2、6、10 Gy;접수6 Gy제량조사후응용자이순1、2、4μg/d간예4주적E、F、G조.우말차급약후2주,처사각조소서병수집고환,계산각조소서고환곡세정관분화지수(tubule differentiation index,TDI),응용역전록-형광정량취합매련식반응검측고환조직억제소βB、Claudin-3、Claudin-11 mRNA적수평,단백질인적법검측고환Claudin-11단백적표체정황. 결과 여A조(100.0)비교,B、C、D조고환TDI현저하강(68.5±6.4、35.0±6.1、16.3±5.7),차이균유통계학의의(P＜0.05),차D조고환조직억제소βB mRNA현저강저(0.5±0.2비1.0±0.1),차이유통계학의의(P＜0.05)；C、D조Claudin-3 mRNA수평분별위A조적2.17배화3.49배,Claudin-11단백수평분별위A조적2.18배화2.23배.여C조(35.0±6.1)비교,F、G조TDI현저증고(61.7±7.2、55.8±11.9),차이균유통계학의의(P＜0.05),억제소βB mRNA수승고,단차이무통계학의의(균P＞0.05)；F、G조Claudin-3 mRNA수평분별위1.3±0.2화1.6±0.3,저우C조(2.2±0.2),차이균유통계학의의(P＜0.05),차F조Claudin-11 mRNA수평(1.2±0.2)화단백표체수평(1.5±0.5)균현저저우C조(1.79±0.2화2.17±0.3),차이균유통계학의의(P＜0.05).TDI여Claudin-3 mRNA、Claudin-11 mRNA수평균정부상관(rs=-0.884,P＜0.05;rs=-0.758,P＜0.05). 결론 γ사선조사가사소서혈고병장조분Claudin-3、Claudin-11표체수평승고.
Objective To investigate the changes of Claudin-3 and Claudin-11,two key components of blood-testis barrier (BTB) on male infertility induced by γ-ray irradiation.Methods Fortytwo KunMing male mice (20-25 g) were divided into one control group,three γ-ray irradiation groups and three estrodiol (E2) intervention groups randomly:Group A,sham controlled; the lower abnominal and scrotal area of the mice in Group B,C,D were irradiated with single dose of 2,6 or 10 Gy 60Co γ-ray after anaesthetizd; 17β-estradiol intervention were initiated in Group E,F,G after 6 Gy γ-ray irradiation via hypodermic injection for 4w at the dose of 1,2,4 μg/d,respectively.Mice were sacrificed 2 w after the last E2 administration.The tubule differentiation index (TDI) was counted in testis sections.InhibinβB,Claudin-3 and Claudin-11 transcription levels were assayed with semiquantitative real time PCR.Claudin-11 protein levels in testis were generated by western blot.Results Compared with sham control group,TDI in three γ-ray irradiation groups were markedly reduced (68.5 ± 6.4,35.0± 6.1,16.3 ± 5.7 vs 100.0,all P＜0.05).InhibinββB mRNA expression level in testis of gourp D was markedly decreased (0.5±0.2 vs 1.0±0.1,P＜0.05).Claudin-3 mRNA levels of group C and D were up-regulated to 2.17 and 3.49 times,respectively.Claudin-11 protein levels were significantly increased to 2.18 and 2.23 times.Compared with group C,TDI in three E2 intervention groups were improved,which were obvious in group F and G (61.7±7.2,55.8±11.9 vs 35.0±6.1,P＜0.05).The InhibinβB mRNA levels were increased,though there were no significant differences (all P ＞0.05).Claudin-3 mRNA levels in group F and G were down-regulated (1.3± 0.2,1.6±0.3 vs.2.2 ± 0.2,all P＜0.05).In group F significantly reduced mRNA level and protein level of Claudin-11 were observed (mRNA:1.2±0.2 vs.1.8±0.2,P＜0.05; Protein:1.5±0.5 vs.2.2±0.3,P＜0.05).It was negatively correlated TDI with mRNA expression levels of Claudin-3 and Claudin-11 in the irradiated testis (rs =-0.884,P＜0.05; rs=-0.758,P ＜0.05,respectively).Conelusions Irradiation could elevated the expression of claudin-3 and claudin-11.