乙醇对小鼠原代肝细胞脂肪酸氧化速度的影响 (pdf)

    【摘要】 目的 建立小鼠肝细胞的体外培养方法，同时检测乙醇对肝细胞脂肪酸β-氧化速度的影响。方法 采用戊巴比妥麻醉小鼠，通过肝脏灌注和胶原酶消化分离小鼠肝细胞，接种于胶原包被的培养皿中进行体外培养。通过HE染色观察细胞的生长状态。在20mmol/L乙醇存在的情况下，通过3H标记的棕榈酸(C16:0)检测肝细胞的脂肪酸氧化速度。结果 成功地分离了肝细胞，肝细胞的存活率≥95%。肝细胞在体外培养48h仍能够保持良好的生长状态。对肝细胞脂肪酸氧化速度的研究发现，20mmol/L酒精能够导致肝细胞的脂肪酸氧化速度降低40%左右，导致肝细胞中脂肪含量增加。结论 本研究成功地建立肝细胞的体外培养方法，采用3H标记的脂肪酸能够敏感地检测脂肪酸氧化速度，而研究发现乙醇能够抑制肝细胞脂肪酸氧化速度。同时，本研究结果为肝细胞的脂肪和糖类代谢的研究提供了实验条件。

    【关键词】 乙醇；肝细胞；脂肪酸

    The effect of ethanol on fatty acid oxidation of mice primary hepatocytes

    ZHAGN Li-jun, SHI Jian-guo,Chen Guang-sheng,et al.Tangdu Hospital of Pathology, Fourth Military Medical Univeristy, Xi’an 710038,China

    【Abstract】 Objective To isolate the mice primary hepatocytes, and the cells were cultured in vitro. The influence of ethanol on fatty acid β-oxidation velocity was measured.Methods The mouse was anesthetized with pentobarbital, and the liver was degested with collagenase. The primary hepatocytes were inoculated in collagen-coated dishes. The morphology of cultured primary hepatocytes was observed by HE staining. The fatty acid β-oxidation was measured using [9,10-3H] palmitic acid (C16:0) as a tracer.Results The primary hepatocytes were isolated and cultured successfully, and the viability was more than 95%. 20mmol/L ethanol could reduce the speed of fatty acid β-oxidation about 40%, and lead to the increased lipid level in hepatocytes.Conclusion We isolated and cultured the mouse primary hepatocytes in vitro, and the influence of ethanol on fatty acid β-oxidation was determined. This study is helpful for the research on the lipid metabolism in liver. ......