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《Chinese Journal of Pathophysiology》 2003-10
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Effect of hypoxia on the expression of proliferating cell nuclear antigen and phenotype of cardiac fibroblasts

YU Jun 1, ZHU Miao-zhang 1, PEI Jian-ming 1, CHEN Bao-ying 2, L Shun-yan 1, DONG Ming-qing 3, WEI Qi-ming 1 (? 1?Department of Physiology,? 2?Department of Radiology, Tangd u Hospital, ? 3?Department of Pathophysiology, The Fourth Military Medical University , Xi'an 710032, China)  
AIM:To investigate effect of hypoxia on the expression of proliferating cell nuclear antigen(PCNA) a nd phenotype of cardiac fibroblasts(CFs). METHODS: The purif ied cardiac fibro blasts were cultured and divided randomly into there groups :control group, mode rate hypoxia(MH) group and severe hypoxia(SH) group. After 72 h,MTT method was u s ed to investigate the proliferation of CFs, and the ultrastructure of fibroblast s were observed with transmission electron microscopy The expression of PCNA a n d α-actin in cardiac fibroblasts were measured by the means of immunohistochemi s try and laser scanning confocal microscopy, respectively. RESULTS: MTT A 490 nm value of MH group was significantly higher than that of control group by (18 4±25 0)% ( P 0 05), whereas MTT A 490 nm value of MH group was signific a ntly lower than that of control group by (15 8±25 0)% ( P 0 05) The immunohistochemistry experim ent showed that there was basal PCNA expression in control CFs, and it was incre ased in MH CFs( P 0 05 vs control group), but there was no significant difference between SH and control CFs. Observed with transmission electron microscopy, the control CFs had typical phenotype of fibroblasts: shuttle-shaped, abundant roug h endoplasmic reticulums (RER) and golgi complexes,but few mitochondria or m icromyofilaments in cytoplasm. After MH for 72 h, there appeared lots of micromy o filaments in cytoplasm, but there were little micromyofilaments in SH CFs. Obser ved with laser confocal scanning microscopy, there was lower expression of α-ac tin in CFs of control group, while many regular bundles of micromyofilaments and the expression of α-actin were signifiantly increased ( P 0.05 vs con trol group) in CFs treated with MH. The α-actin expression in severe hypoxia CFs was not sig nificantly different from control group. CONCLUSION: MH made C Fs have the ch aracteristics of myofibroblasts phenotype and enhanced PCNA expression.
【Fund】: 国家自然科学基金资助项目 (No .39970 32 7)
【CateGory Index】: R363
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