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镉诱导长江华溪蟹(Sinopotamon yangtsekiense)心肌细胞凋亡的研究
雷雯雯, 徐 团, 王 兰
山西大学生命科学学院
摘要:
采用急性毒性实验方法, 研究了不同浓度氯化镉(0 、7.25 、14.50、29.00、58.00 和116.00mg/L)处理不同时间(1d 、3d、5d和7d)对长江华溪蟹心肌细胞凋亡的影响。结果表明, 低浓度组及染毒时间较短时镉对心肌的影响不明显, 但随着染毒浓度的增加和时间的延长, 心肌出现空泡变性、玻璃样变性、炎细胞浸润等病理现象, 在染毒7d虽然观察到疑似的凋亡细胞, 但凋亡率较低; 荧光显微镜未观察到心肌细胞典型的凋亡细胞核形态学变化; 琼脂糖凝胶电泳图谱未出现凋亡细胞所特有的梯状条带; 流式细胞术检测结果显示镉处理组细胞凋亡率增加, 但与对照组相比, 无统计学意义( P >0.05); Caspase-3 活性也没有显著性变化( P >0.05) 。因此, 在本实验所选取的浓度和时间范围内, 镉未诱导长江华溪蟹心肌细胞发生细胞凋亡, 这与本课题组前期研究中观察到的肝胰腺、精巢组织在染毒2d、7d发生细胞凋亡的情况明显不同, 这可能与心脏组织的特殊性、不同组织对镉的积累量不同有关。
关键词:  长江华溪蟹, 心肌, 细胞凋亡, 镉
DOI:10.11693/hyhz201202016016
分类号:
基金项目:国家自然科学基金资助项目, 30870267 号, 30970361 号; 山西省自然科学基金资助项目, 2008011069 号
EFFECTS OF CADMIUM ON APOPTOSIS OF CARDIOMYOCYTE IN THE FRESHWATER CRAB SINOPOTAMON YANGTSEKIENSE
LEI Wen-Wen, XU Tuan, WANG Lan
School of Life Science, Shanxi University
Abstract:
The effects of cadmium (Cd) on cardiomyocyte apoptosis in the freshwater crab Sinopotamon yangtsekiense were investigated by acute systemic toxicity testing methods. To verify if apoptosis is one of the cardiomyocyte death pathways upon Cd administration, crabs were exposed to different concentrations of Cd (0, 7.25, 14.50, 29.00, 58.00 and 116.00 mg/L) for 1, 3, 5 and 7 days. The results of hematoxylin and eosin (H.E.) staining assays showed that there were no obvious alterations in cardiac histology of crabs in low concentrations and short-term exposure groups. With increasing Cd concentrations and exposure time, histological abnormalities appeared including vacuolar and vitreous degeneration, and infiltration of inflammatory cells. Apoptosis-like cells were observed, but rare in 7d exposure groups. Besides, the charac-teristic apoptotic morphologic changes and “ladder” pattern weren’t observed by fluorescence microscopy with Hoechst 33258 staining and agarose gel electrophoresis. Flow cytometry assay indicated that the percentage of apoptotic cells were increased with the increasing of exposure time and concentrati ons of Cd compared with control, but there was no signifi-cant difference. Moreover, the activity of Caspase-3 was not affected by Cd treatment after 7 days exposure. However, these findings suggest that apoptosis wasn’t induced by Cd in cardiomyocyte in the present study. While previous studies in our lab showed that Cd induced apoptosis in the hepatopancreas and testis cells of crabs after 2 days and 7 days exposure. In crab cardiomyocyte, the lack of Cd-induced apoptosis may be attributed to different Cd accumulation amount in the three tissues.
Key words:  Sinopotamon yangtsekiense, Cardiomyocyte, Apoptosis, Cadmium
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