Department of Animal Nutrition and Biotechnology, and Fisheries, University of Agriculture in Krakow, Al. Mickiewicza 24/28, 30-059 Krakow, Poland.
Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Al. Mickiewicza 24/28, 30-059 Krakow, Poland.
Int J Mol Sci. 2023 Aug 2;24(15):12344. doi: 10.3390/ijms241512344.
This study presents a simple and cost-effective method for isolating hepatocytes from liver biopsies obtained from healthy and ketotic dairy cows, which can be utilized for studying cellular metabolism, drug toxicity, and hepatocyte-specific gene function and regulation. The expression of hepatocyte marker genes (, , ) was measured and found to be highest at 6 h post-isolation, with a subsequent decrease over time. Cells isolated from ketotic livers exhibited lower expression levels than those from healthy livers. Furthermore, for the functional characterization of ketotic hepatocytes, the cells were exposed to varying doses of zearalenone (ZEA). While doses of 10-50 µM did not affect cell viability, the highest dose of ZEA (100 µM) significantly decreased cell viability, as measured using XTT assay. Additionally, the potential induction of cytochrome P450 A1 (CYP1A1) by ZEA was found. Despite limitations such as a short-term culture, this model provides a useful tool for conducting toxicological research.
本研究提出了一种简单且经济有效的方法，可从健康和酮病奶牛的肝活检组织中分离肝细胞，该方法可用于研究细胞代谢、药物毒性以及肝细胞特异性基因功能和调控。测量了肝细胞标记基因（、、）的表达水平，发现其在分离后 6 小时最高，随后随时间下降。从酮病肝脏中分离的细胞的表达水平低于健康肝脏。此外，为了对酮病肝细胞的功能进行特征描述，将细胞暴露于不同剂量的玉米赤霉烯酮（ZEA）中。虽然 10-50µM 的剂量不影响细胞活力，但最高剂量的 ZEA（100µM）显著降低了细胞活力，这可通过 XTT 测定法进行测量。此外，还发现 ZEA 可能诱导细胞色素 P450 A1（CYP1A1）。尽管存在短期培养等限制，但该模型为进行毒理学研究提供了有用的工具。
