Current Drug Metabolism - Online First

Human hepatic carcinoma cell lines are widely used in vitro to study lipid and xenobiotic metabolism, as well as glucose regulation in both normal and diseased states. However, their validity is often questioned due to variability in protein expression compared to primary human hepatocytes (cHH). This study aimed to quantify protein abundance in various hepatic cell lines versus cHH and human liver tissue homogenate (HLT) using a data-independent acquisition-based total protein approach (DIA-TPA). We compared the global proteome from the whole cell homogenates of HepaRG, HepG2, and Huh7 cell lines with that of cHH and HLT.

Proteins in whole cell homogenates were digested in solution using pressure-cycling technology (PCT). DIA was performed via sequential window acquisition of theoretical mass spectra (SWATH-MS), and MS2 spectra were quantified using Spectronaut™, followed by analysis with TPA.

We identified 2715, 2578, 2874, 2717, and 3083 proteins in HepaRG, HepG2, Huh7, cHH, and HLT, respectively, at a 1% FDR. The global proteome of cHH significantly differed from that of the cancer hepatic cell lines. Among the cell lines, the global and ADME protein profile of HepaRG most closely correlated with cHH, with 89 out of 101 ADME proteins identified. Clinically relevant DMEs from the CYP450 family (CYP2C9, CYP2C19, CYP2D6, and CYP3A4) and the UGT family (UGT1A1, UGT1A3, UGT1A6, UGT2B7, and UGT2B15) were quantifiable in human hepatocytes, human liver tissue, and the HepaRG cell line. The Huh7 cell line exhibited a higher abundance of proteins related to gluconeogenesis and glycolysis compared to other groups.

This study highlights the potential of untargeted global proteomics in detecting differences in protein expression among various hepatic cell lines and provides a comprehensive database to inform the choice of the cell line in future studies.

Shenfu decoction (SFD), a Traditional Chinese Medicine formula, is used in clinical emergencies. Its effects on seawater-induced hypothermia remain unclear. This study investigates the therapeutic mechanisms of SFD in improving the survival of hypothermic rats through metabolomics and gut microbiota analysis.

Hypothermia was induced in rats via seawater immersion. The chemical constituents of SFD were analyzed using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Survival time and rates of low-temperature water-immersed rats were assessed. Rat blood samples were obtained for analysis of hematologic parameters, electrolytes, hepatic and renal function, cardiac injury, and inflammatory cytokines. To investigate the potential mechanism underlying the survival-prolonging effect of SFD on seawater-immersed hypothermic rats, untargeted blood metabolomics and gut microbiota profiling were employed for preliminary screening.

UPLC-Q-TOF-MS identified almost 50 compounds in SFD, and 1.35 g/kg SFD significantly extended the survival time of seawater-induced hypothermia rats by 6 hours. After hypothermic seawater immersion, the levels of red blood cells, hemoglobin, hematocrit, as well as serum calcium, phosphorus, blood urea nitrogen, alkaline phosphatase, total protein, cardiac troponin T, and interleukin-6 were significantly increased. However, pretreatment with 1.35 g/kg SFD in rats markedly decreased these parameters. The induction of hypothermic seawater immersion elevated blood glucose, and the administration of SFD exacerbated this increase in rats. Metabolomic analysis revealed elevated levels of valerenic acid and benzoylmesaconine in the SFD group, suggesting the restoration of metabolic homeostasis. This recovery was associated with modulation of the gut microbiota, notably an enhancement of beneficial genera, such as Enterococcus.

The findings demonstrated that SFD significantly prolonged survival in a rat model of seawater-immersion hypothermia. The protective mechanism involved a dual action: mitigating hypothermia-induced organ damage and hematological disturbances, coupled with restoring metabolic homeostasis and modulating gut microbiota. SFD has been found to possess specifically enriched beneficial bacterial genera, linked to the activation of brown adipose tissue and non-shivering thermogenesis. This study has provided initial evidence for a gut microbiota-metabolism axis mediating SFD's protective effect.

SFD prolonged survival in rats with seawater-induced hypothermia, likely by enhancing thermogenesis and regulating lipid metabolism through gut microbiota changes. The findings highlighted the potential of SFD for hypothermia prevention; however, its exact underlying mechanisms require further validation.

This systematic review aimed to identify, evaluate, and critically analyze pharmacokinetic models of vancomycin in adult populations published in PubMed and EMBASE between 2020 and 2024.

Twenty-two studies were included, describing 24 models characterized by substantial heterogeneity in terms of study populations, methodological design, and covariate selection. Most models were developed in Asia and focused on hospitalized patients, particularly those in intensive care units (ICUs). Data from 2150 patients were analyzed, with an average of 93 patients per model.

The models demonstrated high variability in pharmacokinetic parameters, such as vancomycin clearance (Cl) and volume of distribution (Vd), influenced by factors, such as renal function, weight, age, and comorbidities. The meta-analysis conducted on clearance and interindividual variability in clearance (IIV Cl) revealed high heterogeneity among the analyzed studies. The average vancomycin clearance was 4.23 L/h, with higher values observed in neurosurgical, oncohematologic patients, and those with increased renal function. The volume of distribution showed greater variability in obese patients and those undergoing continuous renal replacement therapy. Creatinine clearance (ClCr) was identified as a significant covariate in 66% of the models, while weight was significant in 33%. Other important covariates included age, sex, serum creatinine, serum urea, and the hospital admission unit. The meta-analysis of Cl and IIV Cl showed high heterogeneity among the studies, with I² values of 0.83 for Cl and 0.98 for IIV Cl, indicating substantial variability.

The limitations of this study included the diversity of the analyzed populations, which made it challenging to assess the model's suitability. While the models showed advances in precision, challenges, such as the lack of external validation and discrepancies in dosing recommendations, remain.

This review paper has highlighted the need to validate models in diverse populations and clinical settings to optimize personalized vancomycin therapy in adults. The findings have highlighted the importance of validating or adapting pharmacokinetic models to the specific characteristics of each hospital population.

The currently available therapies for acute lung injury (ALI), including glucocorticoids, protease inhibitors, and heparin, have limited clinical efficacy and are often associated with significant side effects. Cepharanthine (CEP) has demonstrated effectiveness in treating pulmonary diseases, but its clinical application is restricted by low solubility and poor bioavailability. This study aimed to develop mannosylated cepharanthine-loaded polymeric micelles (MA-CEP-PMs) to improve CEP bioavailability and enhance lung-targeted delivery for the treatment of ALI.

The pharmacokinetics of MA-CEP-PMs in rats were assessed using Ultra-Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (UPLC-Q-TOF-MS). Lung-targeting ability was evaluated through tissue distribution studies and near-infrared imaging. In a rat model of ALI induced by lipopolysaccharide (LPS), anti-ALI effects were assessed via general physiological indicators, Enzyme-Linked Immunosorbent Assay (ELISA), and Western blot analysis. Hematoxylin-eosin (HE) staining was used to examine hepatotoxicity and nephrotoxicity of MA-CEP-PMs in normal rats. Cytotoxicity of the mannosylated polyethylene glycol–poly(lactic-co-glycolic acid) copolymer (MA-PEG-PLGA) on NR8383 cells was evaluated using the Cell Counting Kit-8 (CCK-8) assay. Cellular uptake experiments were performed to determine the targeting ability of MA-PEG-PLGA in NR8383 cells, and the effects of MA-CEP-PMs on inflammatory cytokines were analyzed using ELISA.

MA-CEP-PMs significantly increased the AUC and exhibited better lung targeting ability compared to the unmodified micelles (p < 0.01). In the ALI model, MA-CEP-PMs improved the thymus and spleen indices, decreased the lung wet-to-dry weight ratio (p < 0.05), alleviated model animal damage, and inhibited inflammatory factor and nuclear factor-κB (NF-κB)–related protein levels (p < 0.05). MA-CEP-PMs exhibited no significant hepatotoxicity or nephrotoxicity. MA-PEG-PLGA exhibited low toxicity against NR8383 cells and greater cell uptake, indicating stronger targeting of the lung. MA-CEP-PMs also exhibited more potent anti-inflammatory effects.

This study focused on the short-term therapeutic effects of ALI, whereas the clinical management of lung injury often requires long-term intervention. Future research should therefore assess the long-term efficacy of this delivery system in chronic lung injury, along with determining its safety profile and potential impacts on extra-pulmonary organs. While the involvement of the NF-κB pathway in the anti-inflammatory effects has been confirmed, it remains to be deciphered whether mannose modification synergistically regulates other signaling pathways and what the specific intracellular targets of CEP are, which would require further exploration through detailed molecular biology experiments.

The MA-CEP-PMs significantly improved CEP bioavailability and increased lung targeting. They exhibited good safety and had a significant effect on ALI management.