肝细胞细胞系用于确定镇静剂和阿片类药物的肝毒性,因为这些药物的肝毒性尚未得到很好的表征。这可能会构成威胁,尤其是危重病人,因为他们经常接受高累积剂量的每日镇痛,并且由于潜在的疾病或治疗期间的并发症,他们通常已经有肝功能受损。基于HepG2/C3A细胞的完善的生物传感器用于确定重症监护环境中常用镇静剂和阿片类药物的肝毒性(咪达唑仑,异丙酚,s-ketamin,硫喷妥钠,芬太尼,瑞芬太尼,和舒芬太尼)。孵育时间为2×3天,在细胞培养基或人血浆中具有临床相关(Cmax)和更高浓度(C5×和C10×)的每种药物。之后,我们测量了细胞计数,活力,乳酸脱氢酶(LDH),线粒体脱氢酶活性,细胞色素P4501A2(CYP1A2),和白蛋白合成。所有测试物质都降低了肝细胞的活力,但舒芬太尼和瑞芬太尼的作用更为明显。培养基和血浆中的舒芬太尼以及仅血浆中的瑞芬太尼减少了细胞计数。舒芬太尼和瑞芬太尼还导致细胞培养上清液中LDH的较高值。使用咪达唑仑和s-氯胺酮观察到线粒体脱氢酶活性降低。与较高浓度的舒芬太尼和瑞芬太尼孵育后,血浆中的微量白蛋白合成减少。瑞芬太尼和s-氯胺酮降低CYP1A2活性,而异丙酚和硫喷妥钠增加了它。我们的发现表明,所测试的镇静剂和阿片类药物均无明显的肝毒性。舒芬太尼,瑞芬太尼,s-氯胺酮在体外表现出中度肝毒性作用。这些药物应谨慎对待易感染肝毒性药物的患者,例如,患有预先存在的肝病或肝功能损害作为其基础疾病的一部分的患者(例如,脓毒症中的低氧性肝炎或胆汁淤积性肝功能障碍)。本课题需要进一步的研究,可能使用更复杂的细胞培养模型和全球药物警戒报告,解决使用的细胞模型的局限性:与原代肝细胞相比,由于CYP酶水平低,HepG2/C3A细胞具有较低的代谢能力。然而,虽然测试模型适用于亲代物质,它不是用于代谢物的毒性测试。
A hepatocyte cell line was used to determine the hepatotoxicity of
sedatives and opioids, as the hepatotoxicity of these drugs has not yet been well characterized. This might pose a threat, especially to critically ill patients, as they often receive high cumulative doses for daily analgosedation and often already have impaired liver function due to an underlying disease or complications during treatment. A well-established biosensor based on HepG2/C3A cells was used for the determination of the hepatotoxicity of commonly used
sedatives and opioids in the intensive care setting (midazolam, propofol, s-ketamin, thiopental, fentanyl, remifentanil, and sufentanil). The incubation time was 2 × 3 days with clinically relevant (Cmax) and higher concentrations (C5× and C10×) of each drug in cell culture medium or human plasma. Afterward, we measured the cell count, vitality, lactate dehydrogenase (LDH), mitochondrial dehydrogenase activity, cytochrome P 450 1A2 (CYP1A2), and albumin synthesis. All tested substances reduced the viability of hepatocyte cells, but sufentanil and remifentanil showed more pronounced effects. The cell count was diminished by sufentanil in both the medium and plasma and by remifentanil only in plasma. Sufentanil and remifentanil also led to higher values of LDH in the cell culture supernatant. A reduction of mitochondrial dehydrogenase activity was seen with the use of midazolam and s-ketamine. Microalbumin synthesis was reduced in plasma after its incubation with higher concentrations of sufentanil and remifentanil. Remifentanil and s-ketamine reduced CYP1A2 activity, while propofol and thiopental increased it. Our findings suggest that none of the tested
sedatives and opioids have pronounced hepatotoxicity. Sufentanil, remifentanil, and s-ketamine showed moderate hepatotoxic effects in vitro. These drugs should be given with caution to patients vulnerable to hepatotoxic drugs, e.g., patients with pre-existing liver disease or liver impairment as part of their underlying disease (e.g., hypoxic hepatitis or cholestatic liver dysfunction in sepsis). Further studies are indicated for this topic, which may use more complex cell culture models and global pharmacovigilance reports, addressing the limitation of the used cell model: HepG2/C3A cells have a lower metabolic capacity due to their low levels of CYP enzymes compared to primary hepatocytes. However, while the test model is suitable for parental substances, it is not for toxicity testing of metabolites.