CAJ | 학술논문

目的：探讨二硫化碳（carbon disulfide ，CS2）对大鼠睾丸组织氧化损伤及p53调控细胞凋亡的影响及可能的分子机制。方法清洁级SD雄性大鼠24只，随机分为4组，即空白对照组（Ⅰ）、低浓度组（Ⅱ）、中浓度组（Ⅲ）、高浓度组（Ⅳ），CS2染毒浓度分别为0、50、250和1250 mg／m3。静式吸入染毒，每周5 d ，每天2 h ，共10周，空白对照组仅吸入新鲜空气。染毒结束后记录大鼠体重、睾丸脏器系数、精子计数和精子活动率；苏木精‐伊红（H E ）染色观察其形态学改变；原位末端转移酶标记技术（TUNEL）法检测大鼠睾丸组织细胞凋亡情况；化学比色法测量大鼠睾丸组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH‐Px）活性及丙二醛（MDA）含量；Western blot法检测p53、PARP、Bax和Bcl‐2蛋白表达含量。结果各CS2染毒组的大鼠体重、睾丸脏器系数、精子相对计数及精子活动率均低于空白对照组（均 P＜0.05）；随着CS2染毒浓度的增加，大鼠睾丸组织形态损伤更加明显，中、高浓度组的凋亡指数明显大于空白对照组（均 P＜0.05），抗氧化酶SOD和GSH‐Px活性呈下降趋势而脂质过氧化分解产物 MDA 则明显上升（P＜0.05，P＜0.01）；Western blot检测结果显示，各CS2染毒组p53蛋白表达含量上升（P＜0.01），PARP蛋白仅在高浓度组有明显降低（P＜0.05 ），而Bax与Bcl‐2蛋白各自呈上升和下降趋势，中、高浓度组Bax与Bcl‐2蛋白含量与空白对照组比较差异有统计学意义（P＜0.05，P＜0.01）。结论氧化应激和p53参与了CS2诱导的大鼠睾丸组织细胞凋亡。
목적：탐토이류화탄（carbon disulfide ，CS2）대대서고환조직양화손상급p53조공세포조망적영향급가능적분자궤제。방법청길급SD웅성대서24지，수궤분위4조，즉공백대조조（Ⅰ）、저농도조（Ⅱ）、중농도조（Ⅲ）、고농도조（Ⅳ），CS2염독농도분별위0、50、250화1250 mg／m3。정식흡입염독，매주5 d ，매천2 h ，공10주，공백대조조부흡입신선공기。염독결속후기록대서체중、고환장기계수、정자계수화정자활동솔；소목정‐이홍（H E ）염색관찰기형태학개변；원위말단전이매표기기술（TUNEL）법검측대서고환조직세포조망정황；화학비색법측량대서고환조직중초양화물기화매（SOD）、곡광감태과양화물매（GSH‐Px）활성급병이철（MDA）함량；Western blot법검측p53、PARP、Bax화Bcl‐2단백표체함량。결과각CS2염독조적대서체중、고환장기계수、정자상대계수급정자활동솔균저우공백대조조（균 P＜0.05）；수착CS2염독농도적증가，대서고환조직형태손상경가명현，중、고농도조적조망지수명현대우공백대조조（균 P＜0.05），항양화매SOD화GSH‐Px활성정하강추세이지질과양화분해산물 MDA 칙명현상승（P＜0.05，P＜0.01）；Western blot검측결과현시，각CS2염독조p53단백표체함량상승（P＜0.01），PARP단백부재고농도조유명현강저（P＜0.05 ），이Bax여Bcl‐2단백각자정상승화하강추세，중、고농도조Bax여Bcl‐2단백함량여공백대조조비교차이유통계학의의（P＜0.05，P＜0.01）。결론양화응격화p53삼여료CS2유도적대서고환조직세포조망。
Objective To investigate effects of carbon disulfide (CS2 )on oxidative stress and p53‐mediated apoptosis in male rat’s testes ,and explore its possible molecular mechanisms.Methods Twenty‐four SD male rats were randomly divided into four groups:control group(Ⅰ ) ,low‐concentration group(Ⅱ ) ,medium‐concentration group(Ⅲ )and high‐concentration group (Ⅳ) .Three treatment groups were exposed to CS2 at different concentrations(50 ,250 ,1250 mg/m3 )by inhalation for 10 weeks , with 2 h per day ,5 days per week.Rats of control group inhaled fresh air.Body weight ,organ index ,sperm count and sperm mo‐tility were recorded 10 weeks after CS2 treatment.Histopathological changes of the testes were observed by HE stai‐ning.TUNEL was used to detect apoptosis of the testes.SOD ,GSH‐Px activities and MDA level were measured by chemical colorimetry in the rats.Also ,we used Western blotting to detect the p53 ,PARP ,Bax and Bcl‐2 proteins levels in each group.Results Body weight ,organ index ,sperm count and sperm motility were decreased in CS2 treatment groups ,compared with the control group (P<0.05).The histopathological changes of the testes became more serious with the increase of CS2 concentration.The apoptosis index was significantly higher in middle and high concentration groups than in control group (P<0.05).Meanwhile ,activities of SOD and GSH‐Px were significantly decreased ,and MDA level was significantly increased (P<0.05 ,P<0.01).Western blotting showed that the level of p53 was significantly increased in CS2 treatment groups(P<0.01) . the level of PARP was declined significantly in high‐concentration group(P<0.05).There were significant differences in the levels of Bax and Bcl‐2 between medium and high concentration groups and the control group(P<0.05 ,P<0.01).Conclusion Oxidative stress and p53 are involved in CS2 induced apoptosis in rat’s testes.