State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Hepatobiliary Pancreat Dis Int. 2020 Apr;19(2):129-137. doi: 10.1016/j.hbpd.2019.10.004. Epub 2019 Nov 5.
For its better differentiated hepatocyte phenotype, C3A cell line has been utilized in bioartificial liver system. However, up to now, there are only a few of studies working at the metabolic alternations of C3A cells under the culture conditions with liver failure plasma, which mainly focus on carbohydrate metabolism, total protein synthesis and ureagenesis. In this study, we investigated the effects of acute liver failure plasma on the growth and biological functions of C3A cells, especially on CYP450 enzymes.
C3A cells were treated with fresh DMEM medium containing 10% FBS, fresh DMEM medium containing 10% normal plasma and acute liver failure plasma, respectively. After incubation, the C3A cells were assessed for cell viabilities, lactate dehydrogenase leakage, gene transcription, protein levels, albumin secretion, ammonia metabolism and CYP450 enzyme activities.
Cell viabilities decreased 15%, and lactate dehydrogenase leakage had 1.3-fold elevation in acute liver failure plasma group. Gene transcription exhibited up-regulation, down-regulation or stability for different hepatic genes. In contrast, protein expression levels for several CYP450 enzymes kept constant, while the CYP450 enzyme activities decreased or remained stable. Albumin secretion reduced about 48%, and ammonia accumulation increased approximately 41%.
C3A cells cultured with acute liver failure plasma showed mild inhibition of cell viabilities, reduction of albumin secretion, and increase of ammonia accumulation. Furthermore, CYP450 enzymes demonstrated various alterations on gene transcription, protein expression and enzyme activities.
由于 C3A 细胞系具有更好的肝细胞表型，因此已将其用于生物人工肝脏系统。但是，到目前为止，只有少数研究在肝衰竭血浆培养条件下研究 C3A 细胞的代谢变化，这些研究主要集中在碳水化合物代谢、总蛋白合成和尿素生成上。在这项研究中，我们研究了急性肝衰竭血浆对 C3A 细胞的生长和生物学功能的影响，尤其是对 CYP450 酶的影响。
C3A 细胞分别用含 10%FBS 的新鲜 DMEM 培养基、含 10%正常血浆的新鲜 DMEM 培养基和急性肝衰竭血浆处理。孵育后，评估 C3A 细胞的细胞活力、乳酸脱氢酶漏出、基因转录、蛋白水平、白蛋白分泌、氨代谢和 CYP450 酶活性。
急性肝衰竭血浆组细胞活力下降 15%，乳酸脱氢酶漏出增加 1.3 倍。不同的肝基因转录表现出上调、下调或稳定。相比之下，几种 CYP450 酶的蛋白表达水平保持不变，而 CYP450 酶活性降低或保持稳定。白蛋白分泌减少约 48%，氨积累增加约 41%。
用急性肝衰竭血浆培养的 C3A 细胞显示出细胞活力轻度抑制、白蛋白分泌减少和氨积累增加。此外，CYP450 酶在基因转录、蛋白表达和酶活性方面表现出不同的变化。
