阴道超声辅照对早孕胚胎绒毛细胞凋亡的研究
摘 要目的：探讨不同时间阴道超声辅照，对人早孕胚胎绒毛细胞凋亡的影响，以期为临床阴道超声辅照时限提供新的佐证。方法：选择拟行人工流产的60例健康早孕妇女，分为对照组(未照射)、辐照3min、辐照5min和辐照10min组。应用探头频率5.0-MHz的阴道超声，对孕囊进行不同时间持续辐照。辐照后24小时行人工流产取绒毛。采用末端脱氧核苷酸转移酶介导的原位末端标记法(TUNEL)行细胞凋亡的组织学检测，免疫组织化学方法(S-P法)检测促/抑凋亡基因Bax和Bcl-2蛋白在组织中的表达。结果：对照组早孕绒毛细胞有少量凋亡细胞存在，辐照组凋亡细胞数目增加；辐照3min组出现凋亡细胞数目增多与对照组比较无显著性差异(P >0.05)。随着照射时间延长凋亡细胞数目明显增加，与对照组及辐照3min组比较有明显差异 (P <0.01)。对照组早孕绒毛中促凋亡Bax蛋白少量分布于合体滋养细胞胞浆中，细胞滋养细胞及间质含量极少，照射组细胞滋养细胞及合体滋养细胞Bax蛋白含量均增加，但照射3min及照射5min组与对照组比较Bax蛋白表达无差异(P >0.05)，照射10min组Bax蛋白表达明显增强，与其他三组比较差异有显著性(P <0.01)。抑制凋亡Bcl-2蛋白表达在各组间比较无显著性差异(P >0.05)。结论：阴道超声辐照后增加早孕绒毛细胞的凋亡，且随照射时间的延长，绒毛细胞的凋亡指数逐渐升高，以照射10min组的绒毛细胞凋亡指数最高。阴道超声辐照后早孕胚胎绒毛细胞滋养细胞及合体滋养细胞Bax蛋白表达增加，随着照射时间的延长Bax蛋白表达逐渐增加； Bcl-2蛋白表达减弱，与阴道超声辐照时间长短无关。孕早期的阴道超声辐照3分钟内是安全的，辐照5分钟相对安全，阴道超声的直接辐照最好不要超过10分钟。
关键词阴道超声；绒毛；凋亡
AbstractObjective：To investigate the effect of TVU on the apoptosis of chorionic villi cells in first-trimester pregnancy. Methods：60 healthy women in first-trimester pregnancy were randomly divided into 4 groups according to different exposure time: control group (bland radiation), 3min, 5min and 10min. Gestation sacs were radiated by TVU with 5.0-MHz transducer. The chorionic villi were collected 24 hours after exposure. The apoptosis of chorionic villi cells were analyzed by TUNEL. The expression of Bax and Bcl-2 proteins were analyzed by immunohistochemistry. Results：The apoptosis cells of chorionic villi were increased according to the time of exposure. In contrast with the control group and 3 min groups, the apoptosis cells were increased significantly in 5 minand 10 min groups (P <0.01). There was no difference between control group and 3 min group (P>0.05). The expression the Bax in syncytiotrophoblast and cytotrophoblasts were increased significantly in 10 min group (P <0.01). The expression of Bcl-2 decreased in 3min, 5min and 10min groups in contrast with control group, but there were no difference between the four groups (P >0.05). Conclusions：The apoptosis of chorionic villi cells increased after the exposure to TVU, and the apoptosis index increased according to the time of exposure. The apoptosis index was highest in the 10min group, but there was no difference between control group and 3 min group. The expression of Bax protein in the cytotrophoblasts and syncytiotrophoblasts increased after the exposure to TVU, and it was accoding to the exposure time. The expression of Bcl-2 protein in the cytotrophoblasts decreased after the exposure to TVU, which was independent on the exposure time. In the early pregnancy stage, it is safe to expose the fetus to TUV within 3 minutes. And it is almost safe within 5 minutes. But the time expose to TUV should not longer than 10 minutes.
Key wordstransvaginal ultrasonography；chorionic villi； apoptosis
随着辅助生殖技术的开展，阴道超声广泛用于卵泡监测、取卵及多胎妊娠减胎术等过程中，且趋于早期化、多次化。阴道超声照射作为一种外源性的应激原对生殖细胞及胚胎潜在的生物效应及其安全性越来越受到重视，辐照后对子代组织或器官的发育分化的影响不应忽视。本研究探讨不同时间阴道超声辅照，对人早孕胚胎绒毛细胞凋亡的影响，以期为临床阴道超声辅照时限提供新的佐证。
1对象与方法
1.1对象
2007年7月～2007年12月我院妇科门诊计划外妊娠的早孕患者60例，平均年龄31.3±3岁，平均妊娠天数54.9±3.4天(月经周期和超声检查用于确定孕周)，孕囊平均直径2.3±0.8cm，均见胎芽和原始心管搏动。随机分成四组,对照组是没有接触阴道超声辐照的14例早孕妇女，辐照组为接受阴道超声辐照的46例早孕妇女，按照不同的辐照时间分为辐照3min组12例、辐照5min组18例和辐照10min组16例。, http://www.100md.com(郝翠芳 曲庆兰 张 宁 包洪初 王 雄 王梅梅 庄丽丽)
关键词阴道超声；绒毛；凋亡
AbstractObjective：To investigate the effect of TVU on the apoptosis of chorionic villi cells in first-trimester pregnancy. Methods：60 healthy women in first-trimester pregnancy were randomly divided into 4 groups according to different exposure time: control group (bland radiation), 3min, 5min and 10min. Gestation sacs were radiated by TVU with 5.0-MHz transducer. The chorionic villi were collected 24 hours after exposure. The apoptosis of chorionic villi cells were analyzed by TUNEL. The expression of Bax and Bcl-2 proteins were analyzed by immunohistochemistry. Results：The apoptosis cells of chorionic villi were increased according to the time of exposure. In contrast with the control group and 3 min groups, the apoptosis cells were increased significantly in 5 minand 10 min groups (P <0.01). There was no difference between control group and 3 min group (P>0.05). The expression the Bax in syncytiotrophoblast and cytotrophoblasts were increased significantly in 10 min group (P <0.01). The expression of Bcl-2 decreased in 3min, 5min and 10min groups in contrast with control group, but there were no difference between the four groups (P >0.05). Conclusions：The apoptosis of chorionic villi cells increased after the exposure to TVU, and the apoptosis index increased according to the time of exposure. The apoptosis index was highest in the 10min group, but there was no difference between control group and 3 min group. The expression of Bax protein in the cytotrophoblasts and syncytiotrophoblasts increased after the exposure to TVU, and it was accoding to the exposure time. The expression of Bcl-2 protein in the cytotrophoblasts decreased after the exposure to TVU, which was independent on the exposure time. In the early pregnancy stage, it is safe to expose the fetus to TUV within 3 minutes. And it is almost safe within 5 minutes. But the time expose to TUV should not longer than 10 minutes.
Key wordstransvaginal ultrasonography；chorionic villi； apoptosis
随着辅助生殖技术的开展，阴道超声广泛用于卵泡监测、取卵及多胎妊娠减胎术等过程中，且趋于早期化、多次化。阴道超声照射作为一种外源性的应激原对生殖细胞及胚胎潜在的生物效应及其安全性越来越受到重视，辐照后对子代组织或器官的发育分化的影响不应忽视。本研究探讨不同时间阴道超声辅照，对人早孕胚胎绒毛细胞凋亡的影响，以期为临床阴道超声辅照时限提供新的佐证。
1对象与方法
1.1对象
2007年7月～2007年12月我院妇科门诊计划外妊娠的早孕患者60例，平均年龄31.3±3岁，平均妊娠天数54.9±3.4天(月经周期和超声检查用于确定孕周)，孕囊平均直径2.3±0.8cm，均见胎芽和原始心管搏动。随机分成四组,对照组是没有接触阴道超声辐照的14例早孕妇女，辐照组为接受阴道超声辐照的46例早孕妇女，按照不同的辐照时间分为辐照3min组12例、辐照5min组18例和辐照10min组16例。, http://www.100md.com(郝翠芳 曲庆兰 张 宁 包洪初 王 雄 王梅梅 庄丽丽)