| 摘要: |
| 【目的】探讨葡萄糖对鹅肝细胞增殖能力的影响。【方法】以四川白鹅为供试动物，采用半原位二步胶原酶灌注法采集鹅的肝细胞，之后用含不同浓度（0（对照组，CK），5，20，35 mmol/L）葡萄糖的培养基培养鹅原代肝细胞，培养48 h后用BrdU免疫荧光染色检测BrdU阳性细胞率，BrdU-ELISA法检测肝细胞DNA含量，ELISA法检测Cyclin D1蛋白质量浓度。反转录聚合酶链式反应检测细胞增殖相关基因（Cyclin D1、Cyclin D2和Cyclin D3）的mRNA表达水平。【结果】与对照组比较，20和35 mmol/L葡萄糖可以显著增加BrdU阳性细胞率，并可以明显提高原代肝细胞的DNA含量；5和20 mmol/L葡萄糖对Cyclin D1蛋白质量浓度影响不显著，而35 mmol/L葡萄糖对Cyclin D1蛋白质量浓度有显著促进作用。Cyclin D1、Cyclin D2和Cyclin D3的mRNA表达水平随着葡萄糖浓度的增加而升高，其中35 mmol/L葡萄糖的促进作用明显大于其他处理。【结论】葡萄糖能够促进鹅原代肝细胞的增殖。 |
| 关键词: 葡萄糖 鹅原代肝细胞 细胞增殖 |
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| 基金项目:国家自然科学基金项目（31101712）；高等学校博士学科点专项科研基金项目（201151103120006)；四川农业大学本科生科研兴趣培养计划项目（2013015） |
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| Effect of glucose on cell proliferation of goose primary hepatocytes |
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YE Feng-jiang,HAN Chun-chun,LIU Dan-dan,et al
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| Abstract: |
| 【Objective】This study aimed to detect the effect of glucose on the proliferation capability of the goose hepatocytes.【Method】Goose primary hepatocytes were isolated from Sichuan White Goose using semi-in situ two-step collagenase perfusion isolation procedure and treated with different glucose solutions (0 (CK),5,20,and 35 mmol/L).After 48 h,the rate of BrdU positive cells was measured by BrdU immunofluorescence staining method,the DNA was measured by BrdU-ELISA method,and protein contents of genes involved in cell proliferation were measured by ELISA.Relative mRNA levels of Cyclin D1,Cyclin D2 and Cyclin D3 were also determined by reverse transcription polymerase chain reaction.【Result】Compared with control group,20 and 35 mmol/L glucose solutions significantly increased the rates of BrdU positive cells and promoted the DNA contents of goose hepatocytes.5 and 20 mmol/L glucose solutions had no evident effect on protein content of Cyclin D1,while 35 mmol/L glucose significantly increased the protein content of Cyclin D1.Relative mRNA levels of Cyclin D1,Cyclin D2 and Cyclin D3 increased with the rising of glucose concentration, and the promoting effect at 35 mmol/L glucose was bigger than those of other groups.【Conclusion】Glucose could promote the cell proliferation of goose primary hepatocytes. |
| Key words: glucose goose primary hepatocytes cell proliferation |
