Huang Jian-Min, Huang Tian-Hua, Qiu Huan-Ying, Fang Xiao-Wu, Zhuang Tian-Gang, Liu Hong-Xi, Wang Yong-Hua, Deng Li-Zhi, Qiu Jie-Wen
Department of Cell Biology and Medical Genetics, Shantou University Medical College, Shantou 515031, Guangdong Province, China.
World J Gastroenterol. 2003 Apr;9(4):736-40. doi: 10.3748/wjg.v9.i4.736.
To evaluate the level of sperm chromosome aberrations in male patients with hepatitis B, and to directly detect whether there are HBV DNA integrations in sperm chromosomes of hepatitis B patients.
Sperm chromosomes of 14 tested subjects (5 healthy controls, 9 patients with HBV infection, including 1 with acute hepatitis B, 2 with chronic active hepatitis B, 4 with chronic persistent hepatitis B, 2 chronic HBsAg carriers with no clinical symptoms) were prepared using interspecific in vitro fertilization between zona-free golden hamster ova and human spermatozoa, and the frequencies of aberration spermatozoa were compared between subjects of HBV infection and controls. Fluorescence in situ hybridization (FISH) to sperm chromosome spreads was carried out with biotin-labeled full length HBV DNA probe to detect the specific HBV DNA sequences in the sperm chromosomes.
The total frequency of sperm chromosome aberrations in HBV infection group (14.8 %, 33/223) was significantly higher than that in the control group (4.3 %, 5/116). Moreover, the sperm chromosomes in HBV infection patients commonly presented stickiness, clumping, failure to staining, etc, which would affect the analysis of sperm chromosomes. Specific fluorescent signal spots for HBV DNA were seen in sperm chromosomes of one patient with chronic persistent hepatitis. In 9 (9/42) sperm chromosome complements containing fluorescent signal spots, one presented 5 obvious FISH spots, others presented 2 to 4 signals. There was significant difference of fluorescence intensity among the signal spots. The distribution of signal sites among chromosomes was random.
HBV infection can bring about mutagenic effects on sperm chromosomes. Integrations of viral DNA into sperm chromosomes which are multisites and nonspecific, can further increase the instability of sperm chromosomes. This study suggested that HBV infection can create extensively hereditary effects by alteration genetic constituent and/or induction chromosome aberrations, as well as the possibility of vertical transmission of HBV via the germ line to the next generation.
评估乙型肝炎男性患者精子染色体畸变水平，并直接检测乙型肝炎患者精子染色体中是否存在乙肝病毒（HBV）DNA整合。
采用无透明带金黄仓鼠卵与人类精子体外受精法制备14例受检者（5例健康对照者、9例HBV感染者，其中急性乙型肝炎1例、慢性活动性乙型肝炎2例、慢性迁延性乙型肝炎4例、无症状慢性HBsAg携带者2例）的精子染色体，比较HBV感染组与对照组间畸变精子的频率。采用生物素标记的全长HBV DNA探针，对精子染色体涂片进行荧光原位杂交（FISH），以检测精子染色体中的特异性HBV DNA序列。
HBV感染组精子染色体畸变总频率（14.8%，33/223）显著高于对照组（4.3%，5/116）。此外，HBV感染患者的精子染色体常出现粘连、聚集、不着色等情况，影响精子染色体分析。在1例慢性迁延性乙型肝炎患者的精子染色体中可见HBV DNA特异性荧光信号点。在9个（9/42）含有荧光信号点的精子染色体组型中，1个呈现5个明显的FISH信号点，其余呈现2至4个信号。信号点间荧光强度存在显著差异。信号位点在染色体间的分布是随机的。
HBV感染可对精子染色体产生诱变作用。病毒DNA整合入精子染色体具有多位点、非特异性的特点，可进一步增加精子染色体的不稳定性。本研究提示，HBV感染可通过改变遗传成分和/或诱导染色体畸变产生广泛的遗传效应，以及HBV经生殖细胞垂直传播给下一代的可能性。
