Overcoming gastrointestinal degradation, epithelial impermeability, and hepatic clearance has been the long-sought goal in oral insulin delivery. Xia et al.’s pioneering work in Science Advances achieves a paradigm shift using site-specific adaptive milk-derived nanovesicles (MiNVs). Engineered from bovine milk exosomes and biothiol-responsive liposomes, MiNVs exploit IgG-FcRn transcytosis for efficient intestinal uptake and glutathione-activated release in the liver. This dual-mechanism enables sustained glycemic control and high oral bioavailability in rodent and porcine diabetic models. More than an incremental advance, this represents a transformative leap toward physiological, injection-free insulin therapy.

The creation of ex vivo human liver models has long been a critical objective in academic, clinical, and pharmaceutical research, particularly for drug development, where accurate evaluation of hepatic metabolic dynamics is crucial. We have developed a bioengineered, perfused, organ-level human liver model that accurately replicates key liver functions, including metabolic activities, and protein synthesis, thus addressing some of the limitations associated with traditional liver monolayers, organoids, and matrix-embedded liver cells. Our approach utilizes liver-specific biomatrix scaffolds, prepared using an innovative protocol and fortified with matrix components that facilitate cellular interactions. These scaffolds, when seeded with human fetal liver cells or co-seeded with liver parenchymal and endothelial cell lines, enable the formation of three-dimensional (3D) human livers with enhanced cellular organization. The "recellularized tissue-engineered livers" (RCLs) have undergone various analyses, demonstrating the capability for establishing liver microenvironments ex vivo. Within 7-14 days, the RCLs exhibit evidence of liver differentiation and metabolic capabilities, underscoring the potential for use in drug metabolism and toxicity studies. Although our study represents a significant step forward, we acknowledge the need for direct comparisons with existing models and further research to fully elucidate the spectrum of regenerative responses. The high drug-metabolizing enzyme activity of RCLs, as demonstrated in our study, provides a promising avenue for investigating drug-induced liver injury mechanisms, contributing to a more detailed understanding of early drug discovery processes.

Background: Hyperuricemia is a serious health problem related to not only gout but also cardiovascular diseases (CVDs). Low-dose aspirin was reported to inhibit uric acid excretion, which leads to hyperuricemia. To decrease hyperuricemia-related CVD, this study aimed to identify the risk of hyperuricemia in people taking aspirin.Method: The original data of this cross-sectional study were obtained from the National Health and Nutrition Examination Survey between 2011 and 2018. Participants who filled in the “Preventive Aspirin Use” questionnaire with a positive answer were included in the analysis. Six machine learning algorithms were screened, and eXtreme Gradient Boosting (XGBoost) was employed to establish a model to predict the risk of hyperuricemia.Results: A total of 805 participants were enrolled in the final analysis, of which 190 participants had hyperuricemia. The participants were divided into a training set and testing set at a ratio of 8:2. The area under the curve for the training set was 0.864 and for the testing set was 0.811. The SHapley Additive exPlanations (SHAP) method was used to evaluate the performances of the modeling. Based on the SHAP results, the feature ranking interpretation showed that the estimated glomerular filtration rate, body mass index, and waist circumference were the three most important features for hyperuricemia in individuals taking aspirin. In addition, triglyceride, hypertension, total cholesterol, high-density lipoprotein, low-density lipoprotein, age, race, and smoking were also correlated with the development of hyperuricemia.Conclusion: A predictive model established by XGBoost algorithms can potentially help clinicians make an early detection of hyperuricemia risk in people taking low-dose aspirin.

BackgroundColorectal cancer (CRC) is one of the most frequently diagnosed cancers. Polo-like kinase 1 (PLK1), a member of the serine/threonine kinase PLK family, is the most investigated and essential in the regulation of cell cycle progression, including chromosome segregation, centrosome maturation and cytokinesis. However, the nonmitotic role of PLK1 in CRC is poorly understood. In this study, we explored the tumorigenic effects of PLK1 and its potential as a therapeutic target in CRC.MethodsGEPIA database and immunohistochemistry analysis were performed to evaluate the abnormal expression of PLK1 in CRC patients. MTT assay, colony formation and transwell assay were performed to assess cell viability, colony formation ability and migration ability after inhibiting PLK1 by RNAi or the small molecule inhibitor BI6727. Cell apoptosis, mitochondrial membrane potential (MMP) and ROS levels were evaluated by flow cytometry. Bioluminescence imaging was performed to evaluate the impact of PLK1 on CRC cell survival in a preclinical model. Finally, xenograft tumor model was established to study the effect of PLK1 inhibition on tumor growth.ResultsFirst, immunohistochemistry analysis revealed the significant accumulation of PLK1 in patient-derived CRC tissues compared with adjacent healthy tissues. Furthermore, PLK1 inhibition genetically or pharmacologically significantly reduced cell viability, migration and colony formation, and triggered apoptosis of CRC cells. Additionally, we found that PLK1 inhibition elevated cellular reactive oxygen species (ROS) accumulation and decreased the Bcl2/Bax ratio, which led to mitochondrial dysfunction and the release of Cytochrome c, a key process in initiating cell apoptosis.ConclusionThese data provide new insights into the pathogenesis of CRC and support the potential value of PLK1 as an appealing target for CRC treatment. Overall, the underlying mechanism of inhibiting PLK1-induced apoptosis indicates that the PLK1 inhibitor BI6727 may be a novel potential therapeutic strategy in the treatment of CRC.

Allogeneic blood vessels are regarded as one of the best natural substitutes for diseased blood vessels due to their good vascular compliance and histocompatibility. Since the supply and demand of allograft blood vessels do not always match in time and space, a good preservation scheme for isolated blood vessels is essential. The abdominal aortas of 110 male Sprague–Dawley (SD) rats were randomly divided into three groups, including cold storage group (4°C) (CSG), frozen storage group (FSG) and ambient storage group (25 ± 2°C) (ASG). Seven time points of preservation for 1, 3, 5, 7, 14, 30 and 90 days were set for detection. The changes in vascular physiological function were evaluated by MTT test and vasoconstriction ability detection, and the changes in vascular wall structure were evaluated by the tension tolerance test and pathological staining. The vascular function of CSG was better than FSG within first the 7 days, but the result was opposite since the 14th day. The vascular wall structure, collagen and elastic fibres of vessels, in CSG, showed oedema within 30 days, and continuous disintegration and rupture at 90 days. The vessel wall structure of FSG remained intact within 90 days. The tensile strength of the vessels in CSG was better than that in FSG within 5 days, and there was no statistical difference between the two groups between the 7th and 30th day, and then, the FSG was higher than CSG on the 90th day. Both cold storage and frozen storage could be applied as safe and effective preservation schemes for isolated rat artery within first 30 days. Cold storage is recommended when the storage time is <14 days, and then, frozen storage is better.

BACKGROUND Long-term aspirin treatment was recommended for secondary prevention of cardiovascular and cerebrovascular disease. However, some studies reveal low-dose aspirin (LDA) can raise serum uric acid (SUA) levels. Thus, the purpose of this study was to analyze whether LDA intake is associated with hyperuricemia. MATERIAL AND METHODS Data was collected from the National Health and Nutrition Examination Survey (NHANES) between 2011 and 2018. All participants over 40 years old and who selected "preventive aspirin use" were included in the study. Logistic regression analyses were used to evaluate the relationship between LDA intake and hyperuricemia. The stratified analysis was based on race and estimated glomerular filtration rate (eGFR). RESULTS A total of 3540 participants were included in the study. Of them, 805 (22.7%) took LDA, and 190 (31.6%) had hyperuricemia. There was no significant association between hyperuricemia and LDA intake (OR=1.22, 95% CI: 0.97-1.54) after adjusting for confounding factors. However, further subgroup analysis by age showed a significant association between LDA intake and hyperuricemia (OR=3.44, 95% CI: 1.88-6.27) among those 40 to 50 years of age. After adjusting for confounding factors, the relationship was still significant (OR=2.28, 95% CI: 1.10-4.73); we also found that race (Hispanic American, OR=1.84, 95% CI: 1.11-3.06) and eGFR under 60 mL/min/1.73 m² (OR=1.94, 95% CI: 1.04-3.62) may play important roles in the development of hyperuricemia. CONCLUSIONS LDA does not increase the hyperuricemia risk in people over 40 years. However, those aged between 40 and 50 years, Hispanic American, and with impaired renal function should have careful evaluation during LDA treatment.

Background: Long-term aspirin treatment was recommended for secondary prevention of cardiovascular and cerebrovascular disease. However, some studies reveal low-dose aspirin (LDA) can raise serum uric acid (SUA) levels. Thus, the purpose of this study was to analyze whether LDA intake is associated with hyperuricemia. Material/Methods: Data was collected from the National Health and Nutrition Examination Survey (NHANES) between 2011 and 2018. All participants over 40 years old and who selected "preventive aspirin use" were included in the study. Logistic regression analyses were used to evaluate the relationship between LDA intake and hyperuricemia. The stratified analysis was based on race and estimated glomerular filtration rate (eGFR).Results: A total of 3540 participants were included in the study. Of them, 805 (22.7%) took LDA, and 190 (31.6%) had hyperuricemia. There was no significant association between hyperuricemia and LDA intake (OR=1.22, 95% CI: 0.97-1.54) after adjusting for confounding factors. However, further subgroup analysis by age showed a significant association between LDA intake and hyperuricemia (OR=3.44, 95% CI: 1.88-6.27) among those 40 to 50 years of age. After adjusting for confounding factors, the relationship was still significant (OR=2.28, 95% CI: 1.10-4.73); we also found that race (Hispanic American, OR=1.84, 95% CI: 1.11-3.06) and eGFR under 60 mL/min/1.73 m2 (OR=1.94, 95% CI: 1.04-3.62) may play important roles in the development of hyperuricemia.Conclusions: LDA does not increase the hyperuricemia risk in people over 40 years. However, those aged between 40 and 50 years, Hispanic American, and with impaired renal function should have careful evaluation during LDA treatment.

The impact of critical illness on patients is profound, resulting in physical, mental, and social consequences and poor health-related quality of life (HRQOL). Several studies investigated HRQOL among patients discharged from the intensive care unit (ICU). However, few systematic reviews cover studies conducted in the last decade or using valid instruments for measuring HRQOL in general ICU survivor populations. Herein, we conduct a systematic review of these studies that followed PRISMA guidelines. We will search PubMed, Web of Science, CINAHL (Cumulative Index to Nursing and Allied Health Literature), Cochrane Library, and Open Grey for papers. We will search for articles reporting the HRQOL of ICU survivors that were written in English and published from 01 January 2012 onward from the date of this protocol's publication. We will also extract HRQOL data and analyze associate factors. The risk of bias will be measured with a standard quality assessment tool. The strength of the results will depend on the number of studies and the consistency of their results. Trial registration number: PROSPERO CRD 42022304279.

Background: Colorectal cancer (CRC) is one of the most common gastrointestinal malignancies, ranking third in morbidity and second in mortality among all cancers in the world. The dysregulation of acid ceramidase (ASAH1) which is involved in sphingolipid metabolism, is known to promote tumorigenesis. As an inhibitor of ASAH1, carmofur (1-hexylcarbamoyl-5-fluorouracil) is used as a chemotherapeutic agent clinically and plays a critical role in ASAH1-induced tumorigenesis. The aim of this study was to explore targeting ASAH1 as a new therapeutic mechanism for CRC.Methods: Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) assay were performed to detect protein expression levels and transcript expression levels. Lentivirus-mediated shRNA (short hairpin RNA) was used to silence ASAH1 and establish ASAH1-knockdown cell lines. Thiazolyl Blue Tetrazolium Bromide (MTT) assay, colony formation and transwell assay were performed to assess cell viability, colony formation ability, migration and invasion ability. Xenograft tumor model was established to study the therapeutic effect of ASAH1 inhibition on tumor growth.Results: Firstly, ASAH1 expression in CRC tissues was higher than normal tissues. In addition, ASAH1 inhibition decreased the proliferation of CRC cells and protected mice from the onset and progression of CRC in the xenograft tumor model. Moreover,ASAH1 was possibly associated with autophagy activation in DLD-1 cells. Conclusions: ASAH1 played a key role in regulating tumorigenesis and cell autophagy in CRC. Therefore, ASAH1 has the potential to be developed as a potential therapeutic target for preclinical studies in CRC.

ObjectiveTo investigate the effect of Shuganning injection (SGN) in alleviating drug-induced cholestasis and the possible mechanisms involved. MethodsThe liver of Sprague-Dawley rats was decellularized to prepare collagen scaffolds, and then the scaffolds were recellularized with human HepG2 cells to obtain the tissue-engineered liver (normal control group). The tissue-engineered liver was perfused with 10 μmol/L chlorpromazine (CPZ) and bile salt mixture to establish a model of drug-induced cholestasis (CPZ group), and the model was further treated with Shuganning injection (103-fold dilution) as the injury protection group (SGN+CPZ group). The markers for hepatocellular injury ［alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and alkaline phosphatase (ALP)］ and the antioxidant and oxidative stress markers ［glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), and reactive oxygen species (ROS)］ were measured for all groups, and the normal control group, the CPZ group, and the SGN+CPZ group were compared in terms of the mRNA and protein expression levels of the enzymes associated with liver bile salt metabolism and the enzymes associated with hepatic cholestasis. HE staining was performed to observe liver pathology. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the CPZ group, the SGN+CPZ group had significant reductions in the markers for hepatocellular injury ALT, AST, LDH, and ALP (all P＜0.000 1), significant increases in the oxidative stress markers GSH and SOD (P＜0.000 1 and P＜0.001), and significant reductions in the markers MDA and ROS (P＜0.000 1 and P＜0.001). Compared with the CPZ group, the SGN+CPZ group had significant reductions in the mRNA expression levels of cholesterol 7α-hydroxylase (CYP7A1) and sterol 12α-hydroxylase (CPY8B1) in hepatocytes (all P＜0.001) and significant increases in the mRNA expression levels of farnesoid X receptor (FXR), small heterodimeric partner (SHP), bile salt export pump (BSEP), and multidrug resistance-associated protein 2 (MRP2) (P＜0.000 1, P＜0.01, P＜0.000 1, and P＜0.000 1). HE staining showed that compared with the CPZ group, the SGN+CPZ group had a significant reduction in hepatocyte injury and a significant increase in the number of cells. ConclusionShuganning injection can alleviate drug-induced cholestatic liver injury caused by chlorpromazine, and it exerts a protective effect by activating FXR in hepatocytes and increasing the expression of SHP to regulate bile salt balance. It also inhibits CYP7A1 and CYP8B1 to reduce the synthesis of hydrophobic bile acids and upregulates the expression of BSEP and MRP2 to promote the excretion of bile salts.

Pancreatic cancer has an extremely terrible prognosis and is a common cause of cancer death. In this study, the clinic value, biological function and underlying mechanisms of Zinc finger protein 655 (ZNF655) in human pancreatic cancer were evaluated. The expression level of ZNF655 in pancreatic cancer was determined by immunohistochemistry (IHC) staining. The biological effects of ZNF655 in pancreatic cancer cells was investigated by loss/gain-of-function assays in vitro and in vivo. The downstream molecular mechanism of ZNF655 was explored using co-immunoprecipitation (Co-IP), dual-luciferase reporter and chromatin immunoprecipitation (Ch-IP). ZNF655 expression was significantly elevated in human pancreatic cancer and possessed clinical value in predicting poor prognosis. Functionally, ZNF655 knockdown inhibited the biological progression of pancreatic cancer cells, which was characterized by weaken proliferation, enhanced apoptosis, arrested cell cycle in G2, impeded migration, and suppressed tumor growth. Mechanistically, ZNF655 played an important role in promoting the binding of E2F transcription factor 1 (E2F1) to the cyclin-dependent kinase 1 (CDK1) promoter. Furthermore, knockdown of CDK1 alleviated the promoting effects of ZNF655 overexpression in pancreatic cancer cells. The promotive role of ZNF655 in pancreatic cancer via CDK1 was determined, which drew further interest regarding its clinical application as a promising therapeutic target.

Context Kuhuang (KH) injection is a widely used anticholestatic drug in the clinic and the mechanisms are still unclear. Objective This study uses a new 3D tissue-engineered (TE) liver platform to study the ability of kuhuang to ameliorate liver injury induced by chlorpromazine (CPZ) and the possible mechanisms involved. Materials and methods The TE livers (n = 25) were divided into 5 groups (n = 5 livers/group) as 3D, 3D + CPZ, 3D + CPZ + KH, 3D + CPZ + GW9662 (a PPAR gamma inhibitor) and 3D + CPZ + KH + GW9662. The treatments with kuhuang (1 mg/mL) and GW9662 (10 mu mol/L) were given to the desired groups on the 7th day of the experimental process. 20 mu mol/L CPZ was added on the 8th day. Results According to the 2D experimental results, the minimum effective concentration of kuhuang is 10 mu g/mL and the optimal effective concentration is 1 mg/mL. Kuhuang ameliorated tissue damage in the TE livers both in terms of tissue structure and culture supernatant. Kuhuang significantly reduced TBA accumulation (38%) and downregulated CYP7A1 (38%) and CYP8B1 (79%). It reduced hepatic levels of ROS (14%), MDA (27%) but increased the levels of GSH (41%), SOD (12%), BSEP (4.4-fold), and MRP2 (74%). Moreover, kuhuang downregulated DR5 (99%) but increased the mRNA expression of PPAR gamma (4-fold). Molecular docking analyses determined the bioactivity of the active compounds of kuhuang through their specific bindings to PPAR gamma. Conclusions Kuhuang could alleviate CPZ-induced cholestatic liver injury by activating PPAR gamma to reduce oxidative stress. Applying kuhuang for the treatment of CPZ-induced liver injury could be suggested.

药物性胆汁淤积(drug induced cholestasis,DRIC)是药物性肝损伤(drug induced liver injury,DILI)的一种常见表现,其病理生理特点是胆汁酸从肝细胞到肠道的转运过程被阻断,使有毒的胆汁酸在肝细胞中积累,产生氧化应激反应,导致其炎症的发生、凋亡和纤维化.DRIC的分子机制自20世纪80年代以来一直在研究,但仍存在争议,其原因之一是引起DRIC的机制研究受制于体外模型的开发,因此,体外模型的开发是研究DRIC机制的关键步骤与瓶颈.由于动物的种属差异性问题,目前诸多研究均集中在开发和进一步优化细胞体外培养模型,以用来预测体内DRIC的发生.本文对DRIC的体外模型作一概述.

Background: Liver transplantation (LT) is a life-saving treatment for patients with end-stage liver disease and acute liver failure.However, in-hospital death cannot be avoided.We designed this study to analyze patients' in-hospital mortality rate after LT and the factors correlated with in-hospital death. Methods:The data of patients who received LT in our hospital between January 11, 2015, and November 19, 2019, were obtained from the China Liver Transplant Registry and medical records.The in-hospital mortality rate was calculated, and factors related to mortality, cause of death, and factors related to cause of death were analyzed by reviewing patients' data.Results: A total of 529 patients who underwent cadaveric LT were enrolled in this study.Modified piggyback orthotopic LT was performed for all patients.Seventy patients died in the hospital after LT, and the in-hospital mortality rate was 13.2%.Factors including model for end-stage liver disease (MELD) score, Child-Pugh grading, intraoperative blood loss, and anhepatic phase were correlated with in-hospital death.MELD score and intraoperative blood loss were determined as the two independent risk factors of in-hospital death.The first two causes of death were infection (34.3%) and primary non-function (15.7%).Pulmonary fungal infection was the main cause of infectious death.MELD score was the independent risk factor for infectious death, and both body mass index of donors and cold ischemic time were independent risk factors of primary non-function.Conclusions: In-hospital death poses a threat to certain patients undergoing LT.Our study suggests that the main cause of in-hospital death is an infection, followed by primary non-function.

AbstractBackgroundLeft‐sided portal hypertension is usually found in patients undergoing pancreaticoduodenectomy (PD) with spleno‐mesenterico‐portal (S‐M‐P) confluence resection. This study is to explore the outcomes of S‐M‐P confluence reconstruction after resection by using bifurcated allogeneic vein.MethodsClinicopathologic data of patients who underwent extensive PD with S‐M‐P confluence resection for carcinoma of pancreatic head/uncinate process in our hospital between December 2011 and August 2018 were retrospectively reviewed and clinical outcomes of vein reconstruction after resection were analyzed.ResultsOf the 37 patients enrolled, S‐M‐P reconstruction by bifurcated allogeneic vein was performed in 24 cases (group 1) and simply splenic vein ligation in 13 cases (group 2). Items including pathological results, blood loss, and complications were comparable between the two groups, operation time was longer in group 1 (573.8 vs. 479.2 min, p = 0.018). Significantly decreased platelet count (205.9 vs. 133.1 × 109/L, p = 0.001) and increased splenic volume (270.9 vs. 452.2 ml, p < 0.001) were observed in group 2 at 6 months after operation. The mean splenic hypertrophy ratio was 1.06 in group 1 and 1.63 in group 2, respectively (p < 0.001). There were four patients with varices were found in group 2, none in group 1.ConclusionsWithout increased complications, reconstructing S‐M‐P confluence by bifurcated allogeneic vein after resection may help to avoid left‐sided portal hypertension.

Granzyme B–producing B cells have been reportedly reported to be an important regulatory B cell subset in the pathogenesis of many diseases. However, its role in liver transplant recipients with acute rejection has never been well elucidated. Seventeen liver transplant recipients with acute rejection and 25 controls with stable liver function were enrolled in this study to determine the function of granzyme B–producing B cells via flow cytometry. Liver transplant recipients with acute rejection had higher expression of granzyme B in CD19+B cells when compared with controls. Following rejection, the granzyme B production was even elevated although comparison failed to reach significance. The percentages of CD27+granzyme B+CD19+B cells and CD138+granzyme B+CD19+B cells were lower than those of CD27−granzyme B+CD19+B cells and CD138−granzyme B+CD19+B cells in patients with acute rejection, respectively. While the production of CD27 and CD138 was not different between liver transplant recipients with and without acute rejection but increased expression of the two surface markers was observed following rejection. Furthermore, the frequency of granzyme B+CD19+B cells correlated with the level of alanine aminotransferase instead of tacrolimus. CD19+B cells could produce granzyme B when stimulated with IgG + IgM and CpG in the presence of the supernatant of activated CD4+CD25−T cells. In return, granzyme B+CD19+B cells could suppress the proliferation of CD4+CD25−T cells in a granzyme B– and contact-dependent manner. The increased expression of granzyme B in CD19+B cells from liver transplant recipients with acute rejection might function as a feedback loop for the activation of the CD4+CD25−T cells.

ObjectiveTo investigate the ability of Ganshuang granule (a liver-protecting drug widely used in clinical practice) extract to reduce N-acetyl-p-aminophenol (APAP)-induced hepatotoxicity and possible mechanisms. MethodsA total of five cell culture groups were set up in this experiment, i.e., normal control group, APAP injury group, and three injury protection groups treated with different concentrations of Ganshuang granule extract. Then 20 mmol/L APAP was added to the cell culture medium and incubated for 24 hours to establish an in vitro model of drug-induced liver injury, and the injury protection groups were treated with different concentrations of Ganshuang granule extract (0.2, 1, and 5 μg/ml) in advance for 8 hours of incubation before APAP were added for 24 hours. Related markers were measured, including the markers for hepatocellular injury ［alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH)］, the markers for mitochondrial injury ［mitochondrial membrane potential, and glutamate dehydrogenase (GDH)］, and antioxidant and oxidative stress markers ［glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS)］. Related mechanism was discussed based on the experimental results. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsGanshuang granule extract alleviated APAP-induced hepatotoxicity, improved cell viability (P＜0001), and reduced the levels of AST, ALT, and LDH in supernatant (P＜0.001, P＜0.001, and P＜0.05). Ganshuang granule extract inhibited APAP-induced hepatocellular oxidative stress, and compared with the APAP group, the Ganshuang granule extract groups had significant reductions in the oxidative stress indicators ROS and MDA (both P＜0.01). Ganshuang granule extract alleviated the loss of mitochondrial membrane potential induced by APAP (P＜0.05) and reduced the content of the mitochondrial injury marker GDH in supernatant (P＜0.001) in a dose-dependent manner. Ganshuang granule extract inhibited the expression of CYP2E1/1A2 (both P＜0.05) and increased the expression of phase Ⅱ enzymes in hepatocytes. Ganshuang granule extract induced the expression of Nrf2 and its downstream genes NQO-1 and GCLC (all P＜0.05). ConclusionGanshuang granule extract can prevent APAP-induced hepatocellular injury through two ways. The first way is that Ganshuang granule extract downregulates the expression of CYP2E1/1A2 and thus reduces the production of NAPQI, a toxic product of APAP; the second way is that Ganshuang granule extract upregulates the expression of the detoxification pathway, which can activate Nrf2 to increase the expression of antioxidant enzymes (SOD and GSH) and phase Ⅱ enzymes and thus accelerate the harmless metabolism of APAP.

目的 基于网络药理学和分子模拟对接技术探究苦黄颗粒治疗自身免疫性肝炎(autoimmune hepatitis,AIH)的活性成分和作用机制.方法 通过中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)获取苦黄颗粒中5味药材的化学成分和作用靶点,并重新纳入药典标记的关键成分;通过PubChem数据库和String平台获得活性化合物的作用靶点,同时采用GeneCards和DisGeNET数据库获得AIH相关基因,两者取交集获得共有靶点,并经Cytoscape软件将结果可视化,通过网络拓扑算法获得核心化合物和核心靶点;通过String平台构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络获得苦黄颗粒治疗AIH的重要靶点,通过DAVID平台对重要靶点进行GO富集及京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析;使用Autodock Vina软件进行分子对接,验证网络分析结果.结果 苦黄颗粒的主要活性成分84个,其中羽扇豆碱、梨果仙人掌基质以及槐胺碱等可能是苦黄颗粒治疗AIH的核心化合物.筛选出苦黄颗粒治疗AIH的7个核心靶点,即IL-6、IL-10、CXCL8、IL-4、ICAM1、HMOX1和IL-2.GO功能富集分析和KEGG通路富集分析显示,苦黄颗粒主要对细胞凋亡进程、炎症反应以及机体免疫等生物过程进程调控.结论 苦黄颗粒可通过多组分-多靶点-多通路调控机体炎症,调节免疫反应,从而发挥治疗AIH的作用,其中7个主要核心靶点可能为苦黄颗粒发挥作用的关键靶点,TNF信号通路、T细胞受体信号通路及Toll样受体信号通路可能是苦黄颗粒参与调控机体炎症和免疫的关键信号通路,可成为后续探究苦黄颗粒治疗AIH作用机制的潜在研究方向.

目的 探讨肝爽颗粒主要成分柚皮苷是否具有预防药物性肝损伤的功能以及保护作用的机制.方法 选取具有肝毒性的药物(曲格列酮和双氯芬酸钠)制造体外模型,预防性地应用不同浓度的柚皮苷,通过半数抑制率(IC50)、转氨酶水平、乳酸脱氢酶水平、Caspase3基因水平评价HL-7702细胞的损伤情况.评价不同浓度的柚皮苷对细胞色素P450(CYP450)家族基因的改变情况.结果 柚皮苷能够明显升高曲格列酮和双氯芬酸钠的IC50,具有降低转氨酶、Caspase3表达的作用.但这种改善具有剂量依赖性,其中100μmol/L柚皮苷的保护作用最明显,1μmol/L柚皮苷和10μmol/L柚皮苷的保护作用不明显.比较CYP代谢酶基因改变情况发现,100μmol/L柚皮苷能明显抑制其表达(P<0.05),1μmol/L柚皮苷和10μmol/L柚皮苷的抑制作用不明显.结论 肝爽颗粒主要成分柚皮苷能够通过抑制CYP450酶的功能,从而起到预防作用.

药物性肝病又称药物性肝损伤,是我国非感染性肝病的第二大病种,美国肝衰竭/肝移植的首要原因.药物性肝损伤主要分为固有型肝损伤和非特异质型肝损伤.固有型药物性肝损伤在临床前安全研究中是无法被预测到的,而大多数药物引起的非特异质型肝毒性在临床前研究中就会被发现从而不会被应用到临床,但其中一个例外就是对乙酰氨基酚(扑热息痛,APAP),这种药物在治疗剂量之内是安全的,但是过量服用则会导致严重的肝损伤甚至急性肝衰竭.在欧美等发达国家,APAP过量服用是导致急性肝衰竭的主要原因.因此进一步阐明APAP引起肝损伤的细胞和分子机制,尤其是肝细胞死亡的机制,有助于尽早识别发生急性肝衰竭和不良预后的危险因素,从而有针对性的开发阻断肝损伤进程的治疗靶点与方案.