AIM: To observe changes of mitochondria and investigate the effect of ethanol on mitochondrial perme- ability transition pore (PTP), mitochondrial membrane potential (MMP, ΔΨm) and intracellular calcium concentratio...AIM: To observe changes of mitochondria and investigate the effect of ethanol on mitochondrial perme- ability transition pore (PTP), mitochondrial membrane potential (MMP, ΔΨm) and intracellular calcium concentration in hepatocytes by establishing an animal model of alcoholic liver disease (ALD). METHODS: Fourty adult male Wistar rats were randomly divided into two groups, the model group (20) was administered alcohol intragastrically plus an Oliver oil diet to establish an ALD model, and the control group (20) was given an equal amount of normal saline. The ultramicrostructural changes of mitochondria were observed under electron microscopy. Mitochondria of liver was extracted, and patency of PTP, mitochondrial membrane potential (ΔΨm), mitochondrial mass and intracellular calcium concentration of isolated hepacytes were detected by flow cytometry using rhodamine123 (Rh123), Nonyl-Acridine Orange and calcium fluorescent probe Fluo-3/AM, respectively. RESULTS: Membrane and cristae were broken or disappeared in mitochondria in different shapes under electron microscopy. Some mitochondria showed U shape or megamitochondrion. In the model group, liver mitochondria PTP was broken, and mitochondria swelled, the absorbance at 450 nm, A540 decreased (0.0136 ± 0.0025 vs 0.0321 ± 0.0013, model vs control, P < 0.01); mitochondria transmembrane potential (239.4638 ± 12.7263 vs 377.5850 ± 16.8119, P < 0.01) was lowered; mitochondrial mass (17.4350 ± 1.9880 vs 31.6738 ± 3.4930, P < 0.01); and [Ca2+]i was increased in liver cells (7.0020 ± 0.5008 vs 10.2050 ± 0.4701, P < 0.01).CONCLUSION: Chronic alcohol intake might lead to broken mitochondria PTP, decreased mitochondria membrane potential and injury, and elevated intracellular Ca2+ production. Ethanol-induced chondriosome injury may be an important mechanism of alcoholic diseases.展开更多
为深入研究脉冲电场对细胞的作用机制,并进一步推广其在临床上的应用,基于有限元仿真软件建立了考虑胞内细胞器的单细胞五层介电模型,从2维和3维的角度研究分析了脉冲电场诱导细胞发生电穿孔的动态过程。同时,鉴于理论球形细胞与真实细...为深入研究脉冲电场对细胞的作用机制,并进一步推广其在临床上的应用,基于有限元仿真软件建立了考虑胞内细胞器的单细胞五层介电模型,从2维和3维的角度研究分析了脉冲电场诱导细胞发生电穿孔的动态过程。同时,鉴于理论球形细胞与真实细胞结构的差异,建立不规则细胞介电模型,讨论分析脉冲电场作用于理论球形细胞与真实细胞电穿孔效应的差异。仿真结果表明:在1.5 k V/cm、10μs脉冲电场作用下,细胞膜上跨膜电位升高达到穿孔跨膜电位阈值(1 V)后发生穿孔,孔密度急增(达到1016数量级),电导率随着时间快速增加,从而进一步改变跨膜电位的分布,穿孔区域由正对电极的点逐渐往四周扩展,最终穿孔区域达到整个表面积的71.4%。而不规则细胞下的跨膜电位、电导率等分布较复杂,不再具有对称分布性,对于畸形度较大的细胞则必须考虑其形状的影响。仿真结果合理解释了脉冲作用下单细胞电穿孔的作用机制,为进一步分析复杂细胞系统提供理论依据,同时推进了脉冲电场的临床应用。展开更多
基金Supported by Natural Science Foundation of Shandong Province, No. 032050113
文摘AIM: To observe changes of mitochondria and investigate the effect of ethanol on mitochondrial perme- ability transition pore (PTP), mitochondrial membrane potential (MMP, ΔΨm) and intracellular calcium concentration in hepatocytes by establishing an animal model of alcoholic liver disease (ALD). METHODS: Fourty adult male Wistar rats were randomly divided into two groups, the model group (20) was administered alcohol intragastrically plus an Oliver oil diet to establish an ALD model, and the control group (20) was given an equal amount of normal saline. The ultramicrostructural changes of mitochondria were observed under electron microscopy. Mitochondria of liver was extracted, and patency of PTP, mitochondrial membrane potential (ΔΨm), mitochondrial mass and intracellular calcium concentration of isolated hepacytes were detected by flow cytometry using rhodamine123 (Rh123), Nonyl-Acridine Orange and calcium fluorescent probe Fluo-3/AM, respectively. RESULTS: Membrane and cristae were broken or disappeared in mitochondria in different shapes under electron microscopy. Some mitochondria showed U shape or megamitochondrion. In the model group, liver mitochondria PTP was broken, and mitochondria swelled, the absorbance at 450 nm, A540 decreased (0.0136 ± 0.0025 vs 0.0321 ± 0.0013, model vs control, P < 0.01); mitochondria transmembrane potential (239.4638 ± 12.7263 vs 377.5850 ± 16.8119, P < 0.01) was lowered; mitochondrial mass (17.4350 ± 1.9880 vs 31.6738 ± 3.4930, P < 0.01); and [Ca2+]i was increased in liver cells (7.0020 ± 0.5008 vs 10.2050 ± 0.4701, P < 0.01).CONCLUSION: Chronic alcohol intake might lead to broken mitochondria PTP, decreased mitochondria membrane potential and injury, and elevated intracellular Ca2+ production. Ethanol-induced chondriosome injury may be an important mechanism of alcoholic diseases.
文摘为深入研究脉冲电场对细胞的作用机制,并进一步推广其在临床上的应用,基于有限元仿真软件建立了考虑胞内细胞器的单细胞五层介电模型,从2维和3维的角度研究分析了脉冲电场诱导细胞发生电穿孔的动态过程。同时,鉴于理论球形细胞与真实细胞结构的差异,建立不规则细胞介电模型,讨论分析脉冲电场作用于理论球形细胞与真实细胞电穿孔效应的差异。仿真结果表明:在1.5 k V/cm、10μs脉冲电场作用下,细胞膜上跨膜电位升高达到穿孔跨膜电位阈值(1 V)后发生穿孔,孔密度急增(达到1016数量级),电导率随着时间快速增加,从而进一步改变跨膜电位的分布,穿孔区域由正对电极的点逐渐往四周扩展,最终穿孔区域达到整个表面积的71.4%。而不规则细胞下的跨膜电位、电导率等分布较复杂,不再具有对称分布性,对于畸形度较大的细胞则必须考虑其形状的影响。仿真结果合理解释了脉冲作用下单细胞电穿孔的作用机制,为进一步分析复杂细胞系统提供理论依据,同时推进了脉冲电场的临床应用。
