Molecular mechanism of hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments

doi:10.3969/j.issn.2095-4344.2015.49.021

Abstract

Abstract:

BACKGROUND: Human hepatocyte models of hypoxia-reoxygenation injury are established to simulate the ischemia/reperfusion injury of transplanted organ. There have been no research reports addressing the molecular mechanism underlying hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments.
OBJECTIVE: To analyze the molecular mechanism of hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments.
METHODS: Rat hepatocyte models of hypoxia-reoxygenation in vitro were established and then randomly divided into control and hypoxia-reoxygenation groups. The hypoxia-reoxygenation group was also subdivided into three subgroups: hypoxia-reoxygenation-2, 4 and 6 hours (after 3 hours of hypoxia, hepatocytes were given oxygen for 2, 4 and 6 hours respectively). Cell morphology was observed by light microscope, ultrastructural
changes by transmission electron microscope and the change in F-actin microfilament content by confocal laser microscopy. HSP27 and cofilin gene and protein levels were determined by real-time polymerase chain reaction and western blot assay respectively.
RESULTS AND CONCLUSION: Light microscope observations showed that spindle cells and exfoliated cells significantly increased in hypoxia-reoxygenation group. Transmission electron microscope observations showed that in the hypoxia-reoxygenation group, the amount of endoplasmic reticulum significantly decreased, mitochondrial density increased and glycogen disappeared compared with the control group. Confocal laser microscopy observations showed that in the hypoxia-reoxygenation group, F-actin microfilament fluorescence disordered, F-actin microfilament morphology significantly changed, staining intensity significantly attenuated and mean fluorescence intensity was significantly lower than that in control group (P < 0.05). Real-time PCR and western blot detection results showed that HSP27, cofilin gene and protein expression levels in the hypoxia-reoxygenation group were significantly lower than in the control group (P < 0.05). These results demonstrate that hypoxia-reoxygenation affects the correct assembly of F-actin microfilaments and weakens the normal cycle of F-actin microfilaments through inhibiting the protein expression of HSP27 and cofilin, and gene transcription in hepatocytes, thereby changing the skeleton of F-actin microfilaments.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

CLC Number:

R318

Zhang Ya-fei, Wang Jia-zhong, Ji Hong, Lu Hong-wei, Lu Le, Wang Jin-long, Shang Hao, Li Yi-ming. Molecular mechanism of hypoxia-reoxygenation injury to hepatocyte membrane F-actin microfilaments[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(49): 7994-7999.

Figures/Tables 5

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