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目的 研究硒 (Se)和维生素 E(VE)缺乏对大鼠肝细胞的影响。方法 以天然低Se低 VE的克山病病区粮作为饲料进行动物试验 ,与人工半合成低 Se低 VE饲料的实验互相印证 ;用光镜、电镜、流式细胞术 (FCM)、末端脱氧核苷酸转移酶介导的缺口末端标记 (TUNEL)、DNA琼脂糖凝胶电泳 5种方法进行肝细胞凋亡检测。结果 无论天然饲料还是人工半合成饲料实验 ,低 Se低 VE组光镜下均可见较多肝细胞胞浆着色深 ,核浓缩。主要分布在中央静脉周围。电镜检查天然低 Se低 VE组可见较多肝细胞核体积缩小 ,核染色质呈块状聚集在核膜下。用流式细胞术检测 ,天然低 Se低 VE组较对照组肝细胞凋亡百分率明显增加 ,补 Se和 /或 VE使调亡百分率有所下降 ,以联合补充 Se和 VE凋亡百分率下降最明显。人工半合成饲料低 Se低 VE组凋亡百分率较高 ,补 Se和 /或 VE使凋亡百分率有所下降 ,但均未达到显著差异。无论天然饲料还是人工半合成饲料实验 ,低 Se低 VE组均可见较多 TUNEL染色阳性细胞 ,加 Se和 /或 VE可使上述改变的细胞有不同程度减少。DNA琼脂糖凝胶电泳 ,无论天然还是人工半合成饲料实验低 Se低 VE组均可见 DNA梯状图象 ,其余各组未见梯状改变。结论 Se和 VE缺乏可诱导大鼠肝细胞凋亡
Objective To study the effects of selenium (Se) and vitamin E (VE) deficiency on rat hepatocytes. Methods Animal experiments were conducted in Keshan disease area where the natural low Se and low VE were used as foodstuffs, which were verified with the experiments of semi-synthetic low-Se and low-VE feedstuffs. Light microscopy, electron microscopy, flow cytometry (FCM) Nucleotide transferase-mediated nick end labeling (TUNEL), DNA agarose gel electrophoresis 5 methods for detection of hepatocyte apoptosis. Results In both low-Se and low-Se groups, more cytoplasm of hepatocytes were found darker and nuclear concentrated under the light microscope in both natural and artificial feedstuffs. Mainly located in the central vein around. Electron microscopy of natural low-Se low VE group shows more liver cell nucleus size is reduced, nuclear chromatin mass gathered in the nuclear membrane. Flow cytometry showed that the apoptosis rate of hepatocytes in the natural low-Se-low VE group was significantly increased compared with that of the control group, and the percentage of apoptosis was decreased in Se and / or VE-supplemented Se and VE groups . The percentages of apoptosis of artificial low-Se low-VE group were higher than that of artificial Se-fed group, while the percentage of apoptosis was decreased with Se and / or VE supplementation, but no significant difference was found. No matter the natural feed or the artificial semi-synthetic feed, more TUNEL-positive cells could be seen in the low-Se-low-VE group. Se and / or VE increased the above-mentioned changed cells to some extent. DNA agarose gel electrophoresis, whether natural or artificial semi-synthetic feed low Se Se VE group were visible DNA ladder image, the other groups did not see ladder-like changes. Conclusion Se and VE deficiency can induce hepatocyte apoptosis in rats