原位末端标记法观察钉螺细胞凋亡的研究

Observation on apoptosis in Oncomelania hupensisby TUNEL technique

目的使用原位末端标记法观察钉螺细胞凋亡,探讨其在钉螺研究上使用效果和前景。方法经苦楝叶浸提液处理的钉螺和对照钉螺去壳后连续冰冻切片,使用原位末端标记法(TdT-mediated dUTP-biotin nick end labeling assay,TUNEL)和苏木素-伊红(Hematoxylin and eosin,HE)染色观察其细胞凋亡。结果诱导组钉螺细胞凋亡发生在消化腺、足部表皮和足部肌肉纤维之间等部位,对照组凋亡则发生在消化腺和足部表皮部位,两组细胞凋亡值有显著差异(P<0.05);TUNEL法观察和HE染色观察钉螺细胞凋亡,两组凋亡值没有显著差异(P>0.05)。结论 TUNEL法可以运用于钉螺细胞凋亡原位检测。

Thirty snails were divided into two groups for exploring the potential benefit of terminal deoxynucleotide transferasemediated dUTP-biotin nick end labeling assay （TUNEL） in research pertaining to Oncomelania hul）ensis by observ ing its effect in cellar apoptosis. Twenty snails in experimental group were immersed in the leaf extracts of Melia azedarac for 24 hours and soft tissues were separated for frozen section （6 μm）. The cellar apoptosis of snails were observed by TUNEL technique and hematoxylin and eosin （HE） staining. Others served as control group. Results showed that there were clusters of positive signal in both experimental and control groups. Apoptotic cells were detected in digestive gland, foot tegument and muscle of experimental snails and in digestive gland and foot tegument of control group. It was found that apoptosis index of experimental group was significantly higher than that in the control （P〈0.05）. There was no significant difference in apoptosis index between TUNEL technique and HE staining （P〉O. 05）. So we concluded that apoptotic cells and apoptosis index in O. hupensis can be detected using TUNEL technique, which indicates that the technique holds promise in relevant studies.