诊断剂量彩色多普勒超声对早孕胚胎组织细胞安全性的影响
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摘要
目的：检测诊断剂量彩色多普勒超声不同时间照射后对早孕小鼠胚胎组织细胞凋亡及超微结构的影响；检测诊断超声不同频率，相同时间照射后对早孕小鼠胚胎组织细胞凋亡影响的差别。方法：1．应用3.5MHz探头固定持续照射孕鼠腹部，24小时后取出标本，琼脂糖凝胶电泳观察不同照射时间的DNA片段情况及细胞超微结构的变化；2．应用不同频率(3.5MHz和6.5MHz)诊断超声，采用相同时间辐照孕鼠胚胎后，对其影响进行对比研究。结果：1．应用3.5MHz探头固定持续照射孕鼠腹部后，琼脂糖凝胶电泳显示，辐照5min以上各组可见不同程度的凋亡梯型，且随辐照时间的延长，梯型越明显。电镜结果显示，除对照组外，其余各组均可见不同程度的凋亡特征改变：细胞核变形，染色质稀疏；线粒体变形、肿胀、嵴模糊、消失、部分空泡化；内质网网腔扩张，核糖体部分脱落；凋亡小体形成；2．不同频率(3.5MHz和6.5MHz)诊断超声，采用相同时间辐照孕鼠胚胎后，频率为3.5MHz探头照射组，除对照组外，其余3组均出现凋亡梯型，且随辐照时间的延长，凋亡梯型越明显；频率为6.5MHz照射组，15min辐照组出现与前两组有差别的凋亡梯型，30min辐照组凋亡梯型更趋明显。结论：1．频率为3.5MHz诊断剂量彩色多普勒超声持续辐照孕鼠胚胎5min以上，可导致胚胎组织细胞凋亡及超微结构发生改变；随着辐照时间的延长，凋亡细胞数量增加和超微结构的改变程度呈一致性改变。提示：在早孕超声检查时，应尽可能缩短检查时间，以免引起胚胎组织细胞的凋亡和超微结构的改变；2．不同频率诊断超声采用相同时间辐照孕鼠胚胎后，声功率愈大(探头频率愈低)，凋亡愈明显。推测经阴道彩色多普勒超声检查因其声功率低，探头频率高，图像显示更清晰，检测时间短，因而在早期的胚胎检查中相对更安全。
Objective: To observe the effect of mouse embryo cell apoptosis and ultrastructure changes caused by diagnostic dose of color Doppler ultrasound. Methods: 1.Early pregnant mice were irradiated with diagnostic dose of color Doppler ultrasound (3.5MHz). The DNA fragments were electrophoresed in an agarose gel and the finestructure changes were observed by election microscope (EM). 2. Early pregnant mice were irradiated with variant diagnostic dose of color Doppler ultrasound (3.5MHz, 6.5 MHz) and the DNA fragments were electrophoresed in an agarose gel. Results: 1. Apoptic ladders were detected in the 3.5MHz transducer group except the control, as well as the ladder was clearer according to the treated time. Typical ultrastructure changes were seen in the same groups ,which showed rarefaction of nuclear chromosome, patches of condensed chromatin lie against the nuclear membrane, obscureness of swelling mitochondria, crista disappearance and vacuolized, apoptotic body appeared at the same time. 2. Apoptic ladde
rs were detected in the 3.5MHz transducer group except the control, as well as the ladder was clearer according to the treated time. Moreover in the 6.5Mhz transducer group, apoptic ladder was detectable only in 30 min treated with ultrasoundConclusion: Continuous irradiation by diagnostic dose of color Doppler ultrasound may result in mouse embryos cells apoptosis. Frequency of 6.5 MHz maybe more safety than 3.5MHz.
Objective: To observe the effect of mouse embryo cell apoptosis and ultrastructure changes caused by diagnostic dose of color Doppler ultrasound. Methods: 1.Early pregnant mice were irradiated with diagnostic dose of color Doppler ultrasound (3.5MHz). The DNA fragments were electrophoresed in an agarose gel and the finestructure changes were observed by election microscope (EM). 2. Early pregnant mice were irradiated with variant diagnostic dose of color Doppler ultrasound (3.5MHz, 6.5 MHz) and the DNA fragments were electrophoresed in an agarose gel. Results: 1. Apoptic ladders were detected in the 3.5MHz transducer group except the control, as well as the ladder was clearer according to the treated time. Typical ultrastructure changes were seen in the same groups ,which showed rarefaction of nuclear chromosome, patches of condensed chromatin lie against the nuclear membrane, obscureness of swelling mitochondria, crista disappearance and vacuolized, apoptotic body appeared at the same time. 2. Apoptic ladde
rs were detected in the 3.5MHz transducer group except the control, as well as the ladder was clearer according to the treated time. Moreover in the 6.5Mhz transducer group, apoptic ladder was detectable only in 30 min treated with ultrasoundConclusion: Continuous irradiation by diagnostic dose of color Doppler ultrasound may result in mouse embryos cells apoptosis. Frequency of 6.5 MHz maybe more safety than 3.5MHz.
引文
1．冯若．图像诊断技术中的超声诊断发展动态．中国超声医学杂志，1994，10(1)：50-52
2．杜联芳，张青萍，刘望彭。诊断超声对大鼠睾丸生殖细胞凋亡的研究。中国超声医学杂志，2000，16(1)：6-8
3．杜联芳，张青萍，刘望彭。诊断超声与大鼠角膜凋亡。中国超声医学杂志 2000，16(9)：645-647．
4．程颜苓，段云友，曹铁生等。诊断剂量超声与胎鼠中枢神经元的凋亡。中华超声影像学杂志，2001，10(10)：624-626
5. Moore KJ and Matlashewski G. Intracellular infection by Leishmania donovani inhibits macrophage apoptosis. J Immunol, 1994, 152: 2930-2937
6. Mayhew TM, Leach L, Mcgee R, et al. Proliferation, differentiation and apoptosis in Villous traphoblast at 13-41 weeks of gestation(including observation on annulate lamellae and nuclear pore complexes). Placenta, 1999, 20(5-6): 407-422
7. Huppertz B, Reister F, Kingdom J, et al. Apoptosis cascade progresses during turnover of human trophoblast: analysis of Villous cytotrophoblast and syncytial fragments in vitro. Lab Invest, 1999, 79(12): 1687-1702
8. Turner TT, Tung KS, Tomomasa H, et al. Acute testicular ischemia results in germ cell-specific apoptosis in the rat. Biol Repord, 1997, 57(6): 1267-1274
9. Socher, Steven A, Yin, et al. Temperature-mediated germ cell loss in the testis is associated with altered expression of the cell-cycle regulator P53. J Urology, 1997, 157(5): 1986-198
10. Hasegawa M, Wilson G, Russell LD, et al. Radiation-induced cell death in the mouse testis: relationship to apoptosis. Radiat Res, 1997, 147(4): 457-467
11. Taylor MF, Woolverideg I, Metcalfe AD, et al. Leydig cell apoptosis in the rat testes after administration of the cytotoxin ethane dimethanesulphonate: role of the bcl-2 family members. J Endocrinol, 1998, 157(2): 317-326
12. Cardinale A, Lagalla R, De Maria M, et al. Experimental studies on possible biological effects of ultrasonic diagnosis: ultrastructural analysis of rat embry. Radiol Med Torino, 1985,71(7-8): 506-509
13．冯泽平，汤海军，吴连连等。诊断超声对中孕宫内胎儿卵巢超微结构影响的研究。中国超声医学杂志，1997，13(12)：43-46
14．冯泽平，陈辉，吴连连等。诊断超声对中孕宫内胎儿脑垂体超微结构影响的研究。中国超声医学杂志，1998，14(11)：3-5
15. Liu X, Kim CN, Yang J, et al. Induction of apoptotic program in cell-free extracts: requirement or dATP and cytochrome c. Cell, 1996, 86(1): 147-157
16. Susin SA, Zamzami N, Castedo M, et al. Bcl-2 inhibits the mitochondrial release of an apoptogenic protease. J Exp Med, 1996, 184(4): 1331-1341
2．杜联芳，张青萍，刘望彭。诊断超声对大鼠睾丸生殖细胞凋亡的研究。中国超声医学杂志，2000，16(1)：6-8
3．杜联芳，张青萍，刘望彭。诊断超声与大鼠角膜凋亡。中国超声医学杂志 2000，16(9)：645-647．
4．程颜苓，段云友，曹铁生等。诊断剂量超声与胎鼠中枢神经元的凋亡。中华超声影像学杂志，2001，10(10)：624-626
5. Moore KJ and Matlashewski G. Intracellular infection by Leishmania donovani inhibits macrophage apoptosis. J Immunol, 1994, 152: 2930-2937
6. Mayhew TM, Leach L, Mcgee R, et al. Proliferation, differentiation and apoptosis in Villous traphoblast at 13-41 weeks of gestation(including observation on annulate lamellae and nuclear pore complexes). Placenta, 1999, 20(5-6): 407-422
7. Huppertz B, Reister F, Kingdom J, et al. Apoptosis cascade progresses during turnover of human trophoblast: analysis of Villous cytotrophoblast and syncytial fragments in vitro. Lab Invest, 1999, 79(12): 1687-1702
8. Turner TT, Tung KS, Tomomasa H, et al. Acute testicular ischemia results in germ cell-specific apoptosis in the rat. Biol Repord, 1997, 57(6): 1267-1274
9. Socher, Steven A, Yin, et al. Temperature-mediated germ cell loss in the testis is associated with altered expression of the cell-cycle regulator P53. J Urology, 1997, 157(5): 1986-198
10. Hasegawa M, Wilson G, Russell LD, et al. Radiation-induced cell death in the mouse testis: relationship to apoptosis. Radiat Res, 1997, 147(4): 457-467
11. Taylor MF, Woolverideg I, Metcalfe AD, et al. Leydig cell apoptosis in the rat testes after administration of the cytotoxin ethane dimethanesulphonate: role of the bcl-2 family members. J Endocrinol, 1998, 157(2): 317-326
12. Cardinale A, Lagalla R, De Maria M, et al. Experimental studies on possible biological effects of ultrasonic diagnosis: ultrastructural analysis of rat embry. Radiol Med Torino, 1985,71(7-8): 506-509
13．冯泽平，汤海军，吴连连等。诊断超声对中孕宫内胎儿卵巢超微结构影响的研究。中国超声医学杂志，1997，13(12)：43-46
14．冯泽平，陈辉，吴连连等。诊断超声对中孕宫内胎儿脑垂体超微结构影响的研究。中国超声医学杂志，1998，14(11)：3-5
15. Liu X, Kim CN, Yang J, et al. Induction of apoptotic program in cell-free extracts: requirement or dATP and cytochrome c. Cell, 1996, 86(1): 147-157
16. Susin SA, Zamzami N, Castedo M, et al. Bcl-2 inhibits the mitochondrial release of an apoptogenic protease. J Exp Med, 1996, 184(4): 1331-1341