Liver Cholesterol Research Articles

Microbial creation of β-Nicotinamide mononucleotide and its regulation of lipid metabolism in the liver of high-fat diet mice.

β-Nicotinamide mononucleotide (NMN) is a biologically active nucleotide that regulates the physiological metabolism of the body by rapidly increasing nicotinamide adenine dinucleotide (NAD+). To determine the safety and biological activity of NMN resources, we constructed a recombinant strain of P. pastoris that heterologously expresses nicotinamide-phosphate ribosyltransferase (NAMPT), and subsequently catalyzed and purified the expressed product to obtain NMN. Consequently, this study established a high-fat diet (HFD) obese model to investigate the lipid-lowering activity of NMN. The findings showed that NMN supplementation directly increased the NAD+ levels, and reduced HFD-induced liver injury and lipid deposition. NMN treatment significantly decreased total cholesterol (TC) and triglyceride (TG) in serum and liver, as well as alanine aminotransferase (ALT) and insulin levels in serum (p < .05 or p < .01). In conclusion, this study combined synthetic biology with nutritional evaluation to confirm that P. pastoris-generated NMN modulated lipid metabolism in HFD mice, offering a theoretical framework and evidence for the application of microbially created NMN.

Gastric peroral endoscopic myo-neurotomy as less-invasive bariatric surgery in porcine models: an experimental study

Background and AimsObesity is a global health problem, and less-invasive treatments are warranted. We developed a novel endoscopic intervention named peroral endoscopic myo-neurotomy (POEM-N) that is performed at the greater curvature of the gastric antrum as a bariatric/metabolic procedure and assessed its feasibility using porcine models. MethodsSix live pigs were divided into three groups of two: POEM-N with high-fat diet (HFD) (POEM-N group); sham procedure with HFD (HF-control group); and sham procedure with normal diet (N-control group). All pigs were fed the diets for 2 weeks, underwent POEM-N or sham procedure, and were followed up for 2 weeks after the procedure. The primary outcome was the safety and feasibility of POEM-N. Adverse events and the histology of the area after POEM-N were evaluated to assess the procedure’s safety and feasibility. The secondary outcomes were the changes in body weight and glycolipid levels. ResultsThere was no severe worsening of vital signs during or after the POEM-N procedure. All pigs were well and food intake was good, with no delayed adverse events. Excessive storage of dietary residue, deformity of the gastric antrum and fold concentration were observed after POEM-N, as expected. Histological examination showed hypertrophic regeneration of Auerbach’s plexus, which regulates gastric peristalsis, in both POEM-N pigs. After the procedure, HFD-induced weight gain in POEM-N pigs slowed down, and the increase in serum and liver cholesterol level was suppressed. ConclusionsWe applied POEM-N as a bariatric/metabolic procedure and showed its safety and feasibility using porcine models.

Exposure memory and susceptibility to ambient PM2.5: A perspective from hepatic cholesterol and bile acid metabolism

Both epidemiological and experimental studies increasingly show that exposure to ambient fine particulate matter (PM2.5) is related to the occurrence and development of chronic diseases, such as metabolic diseases. However, whether PM2.5 has "exposure memory" and how these memories affect chronic disease development like hepatic metabolic homeostasis are unknown. Therefore, we aimed to explore the effects of exposure transition on liver cholesterol and bile acids (BAs) metabolism in mice. In this study, C57BL/6 mice were exposed to concentrated ambient PM2.5 or filtered air (FA) in a whole-body exposure facility for an initial period of 10 weeks, followed by another 8 weeks of exposure switch (PM2.5 to FA and FA to PM2.5) comparing to non-switch groups (FA to FA and PM2.5 to PM2.5), which were finally divided into four groups (FF of FA to FA, PP of PM2.5 to PM2.5, PF of PM2.5 to FA, and FP of FA to PM2.5). Our results showed no significant difference in food intake, body composition, glucose homeostasis, and lipid metabolism between FA and PM2.5 groups after the initial exposure before the exposure switch. At the end of the exposure switch, the mice switched from FA to PM2.5 exposure exhibited a high sensitivity to late-onset PM2.5 exposure, as indicated by significantly elevated hepatic cholesterol levels and disturbed BAs metabolism. However, the mice switched from PM2.5 to FA exposure retained a certain memorial effects of previous PM2.5 exposure in hepatic cholesterol levels, cholesterol metabolism, and BAs metabolism. Furthermore, 18-week PM2.5 exposure significantly increased hepatic free BAs levels, which were completely reversed by the FA exposure switch. Finally, the changes in small heterodimeric partner (SHP) and nuclear receptor subfamily 5 group A member 2 (LRH1) in response to exposure switch mechanistically explained the above alterations. Therefore, mice switching from PM2.5 exposure to FA showed only a weak memory of prior PM2.5 exposure. In contrast, the early FA caused mice to be more susceptible to subsequent PM2.5 exposure.

Combinatorial metabolomic and transcriptomic analysis of muscle growth in hybrid striped bass (female white bass Morone chrysops x male striped bass M. saxatilis)

BackgroundUnderstanding growth regulatory pathways is important in aquaculture, fisheries, and vertebrate physiology generally. Machine learning pattern recognition and sensitivity analysis were employed to examine metabolomic small molecule profiles and transcriptomic gene expression data generated from liver and white skeletal muscle of hybrid striped bass (white bass Morone chrysops x striped bass M. saxatilis) representative of the top and bottom 10 % by body size of a production cohort.ResultsLarger fish (good-growth) had significantly greater weight, total length, hepatosomatic index, and specific growth rate compared to smaller fish (poor-growth) and also had significantly more muscle fibers of smaller diameter (≤ 20 µm diameter), indicating active hyperplasia. Differences in metabolomic pathways included enhanced energetics (glycolysis, citric acid cycle) and amino acid metabolism in good-growth fish, and enhanced stress, muscle inflammation (cortisol, eicosanoids) and dysfunctional liver cholesterol metabolism in poor-growth fish. The majority of gene transcripts identified as differentially expressed between groups were down-regulated in good-growth fish. Several molecules associated with important growth-regulatory pathways were up-regulated in muscle of fish that grew poorly: growth factors including agt and agtr2 (angiotensins), nicotinic acid (which stimulates growth hormone production), gadd45b, rgl1, zfp36, cebpb, and hmgb1; insulin-like growth factor signaling (igfbp1 and igf1); cytokine signaling (socs3, cxcr4); cell signaling (rgs13, rundc3a), and differentiation (rhou, mmp17, cd22, msi1); mitochondrial uncoupling proteins (ucp3, ucp2); and regulators of lipid metabolism (apoa1, ldlr). Growth factors pttg1, egfr, myc, notch1, and sirt1 were notably up-regulated in muscle of good-growing fish.ConclusionA combinatorial pathway analysis using metabolomic and transcriptomic data collectively suggested promotion of cell signaling, proliferation, and differentiation in muscle of good-growth fish, whereas muscle inflammation and apoptosis was observed in poor-growth fish, along with elevated cortisol (an anti-inflammatory hormone), perhaps related to muscle wasting, hypertrophy, and inferior growth. These findings provide important biomarkers and mechanisms by which growth is regulated in fishes and other vertebrates as well.

Effects of ursodeoxycholic acid on growth performance and lipid metabolism in juvenile large yellow croaker (Larimichthys crocea)

Bile acids (BAs) have been shown to exert beneficial effects on growth and metabolism in various fish species. Nevertheless, few studies have focused on the impact of ursodeoxycholic acid (UDCA). The purpose of the present study was to investigate the potential effects of dietary UDCA supplementation on growth performance and lipid metabolism in juvenile large yellow croaker (Larimichthys crocea). Four diets were formulated based on a control diet (CON), with the addition of 50 mg/kg (UL), 100 mg/kg (UM), and 500 mg/kg (UH) UDCA. A total of 400 juveniles (with an initial body weight of 15.55 ± 0.02 g) were randomly distributed into 16 tanks (1000L) with 25 juveniles per tank. Each type of experimental diet was fed to the fish in four replicates twice daily. After a 10-week feeding trial, the results demonstrated that feeding diets containing 50 and 100 mg/kg UDCA significantly improved growth performance (P < 0.05). Dietary UDCA supplementation significantly increased whole-body crude lipid content, liver, and serum total cholesterol (T-CHO) content, as well as intestinal lipase activity, while reducing liver and serum free fatty acid content (P < 0.05). To investigate the mechanisms underlying the observed growth and metabolic alterations induced by UDCA, tissue samples from the CON and the UM groups were subjected to RNA-sequencing analysis. A total of 1924, 4399, 249, and 971 differentially expressed genes (DEGs) in the liver, intestine, olfactory, and brain tissues were identified, respectively. Enrichment analysis of these DEGs suggested a high degree of pathway similarity between the liver and intestinal tissues. Specifically, the UM diet markedly upregulated genes associated with lipid synthesis and lipid transport, while concurrently downregulating those involved in fatty acid β-oxidation. Weighted gene co-expression network analysis (WGCNA) across tissues revealed a gene cluster related to cholesterol synthesis. Within this cluster, key genes such as 3-hydroxy-3-methylglutaryl-coenzyme A reductase (hmgcr) and 3-hydroxy-3-methylglutaryl-CoA synthase 1 (hmgcs1) were upregulated, implying that UDCA enhances of cholesterol synthesis. Furthermore, in vitro experiments in primary hepatocyte corroborated the role of UDCA in stimulating lipid synthesis, as evidenced by the augmented total triglyceride (TG) and T-CHO levels following UDCA incubation. This increase was paralleled by an upregulation in the expression of genes related to fatty acid and cholesterol synthesis as well as lipid transport. In summary, these results demonstrate that UDCA exerts a beneficial impact on growth performance, lipid transport, and lipid synthesis in juvenile large yellow croakers.

Analysis of Metabolic Changes in Endogenous Metabolites and Diagnostic Biomarkers for Various Diseases Using Liquid Chromatography and Mass Spectrometry.

Analysis of endogenous metabolites in various diseases is useful for searching diagnostic biomarkers and elucidating the molecular mechanisms of pathophysiology. The author and collaborators have developed some LC/tandem mass spectrometry (LC/MS/MS) methods for metabolites and applied them to disease-related samples. First, we identified urinary conjugated cholesterol metabolites and serum N-palmitoyl-O-phosphocholine serine as useful biomarkers for Niemann-Pick disease type C (NPC). For the purpose of intraoperative diagnosis of glioma patients, we developed the LC/MS/MS analysis methods for 2-hydroxyglutaric acid or cystine and found that they could be good differential biomarkers. For renal cell carcinoma, we searched for various biomarkers for early diagnosis, malignancy evaluation and recurrence prediction by global metabolome analysis and targeted LC/MS/MS analysis. In pathological analysis, we developed a simultaneous LC/MS/MS analysis method for 13 steroid hormones and applied it to NPC cells, we found 6 types of reductions in NPC model cells. For non-alcoholic steatohepatitis (NASH), model mice were prepared with special diet and plasma bile acids were measured, and as a result, hydrophilic bile acids were significantly increased. In addition, we developed an LC/MS/MS method for 17 sterols and analyzed liver cholesterol metabolites and found a decrease in phytosterols and cholesterol synthetic markers and an increase in non-enzymatic oxidative sterols in the pre-onset stage of NASH. We will continue to challenge themselves to add value to clinical practice based on cutting-edge analytical chemistry methodology.

Effects of a Functional Food Made with Salvia hispanica L. (Chia Seed), Amaranthus hypochondriacus L. (Amaranth), and an Ethanolic Extract of Curcuma longa L. (Curcumin) in a Rat Model of Childhood Obesity.

Obesity is a global health problem and is increasing in prevalence in most countries. Although obesity affects all age groups, children are the most vulnerable sector. Functional foods are novel formulated foods containing substances (i.e., nutrients, phytochemicals, probiotics, etc.) that have potential health-enhancing or disease-preventing value. The research objective was to study the possible beneficial effects of providing a functional food made with amaranth flour, chia seed, and curcumin extract on the metabolism and behavior of a rat model of childhood obesity. Male Wistar rat pups from two litters of different sizes, a normal litter (NL) (10 pups) and a small litter (SL) (4 pups), were used. After weaning, the rats were fed a hypercaloric diet (HD) or an HD supplemented with the functional food mixture. Body weight and energy intake were measured for seven weeks, and locomotor activity, learning, and memory tests were also performed. At the end of the experiment, glucose and lipid metabolism parameters were determined. The results showed that in this model of obesity produced by early overfeeding and the consumption of a hypercaloric diet, anxiety-like behaviors and metabolic alterations occurred in the rat offspring; however, the provision of the functional food failed to reduce or prevent these alterations, and an exacerbation was even observed in some metabolic indicators. Interestingly, in the NL rats, the provision of the functional food produced some of the expected improvements in health, such as significant decreases in body weight gain and liver cholesterol and non-significant decreases in adipose tissue and leptin and insulin serum levels.

Regulatory Mechanism of Xiaozheng Tongluo Method on Hepatic Lipid Metabolism in ApoE-/- Mice: Insights from the Sterol Regulatory Element-binding Protein-2/3-hydroxy-3-methylglutaryl Coenzyme A Reductase/Low-density Lipoprotein Receptor Signaling Pathway

Abstract Objective: The objective of the study was to explore the regulatory mechanism of the Xiaozheng Tongluo method on lipid metabolism in liver tissue based on the sterol regulatory element-binding protein-2/(SREBP2)/3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR)/low-density lipoprotein receptor (LDLR) signaling pathway. Materials and Methods: C57BL/6 mice were selected as the blank group. The apolipoprotein E-deficient (ApoE-/-) mice were randomly divided into the model group, traditional Chinese medicine (TCM) group, and control group. The general condition of the mice was determined by the mouse’s state and liver weight. Liver hematoxylin and eosin staining and Oil Red O staining were used to observe the pathological changes and lipid droplet deposition of the liver. The protein expression levels of SREBP2, HMGCR, and LDLR were detected by Western blotting and polymerase chain reaction. Results: (1) The rats in the model group were in poor condition, and their liver weight increased significantly. Compared with the model group, the condition of the TCM group and the control group improved to varying degrees, and their liver weight decreased significantly. (2) Compared with the normal group, the hepatocytes in the model group were arranged in a disorderly manner, and the red-stained lipids of stem cells increased significantly. Compared with the model group, the degree of liver lesions in the control group and TCM group was reduced, and the red-stained lipid of hepatocytes was significantly reduced. (3) Compared with the blank group, the expression of SREBP2 and HMGCR protein in the model group increased significantly, and the expression of LDLR protein decreased significantly (P &lt; 0.05). Compared with the model group, the expression of SREBP2 and HMGCR protein in the TCM group decreased significantly, and the expression of LDLR protein increased significantly (P &lt; 0.05). Conclusions: The method detailed in this paper can increase the expression of SREBP2 and HMGCR protein and decrease the expression of LDLR protein, thus regulating liver cholesterol metabolism and delaying the progression of atherosclerosis.

A novel compounded cyproheptadine hydrochloride oral solution: Accelerated stability study and effects on glucose and lipid metabolism in Wistar rats

The present study developed a compounded oral solution (COS) containing an active component cyproheptadine hydrochloride (CY) in association with pharmaceutical adjuvants. The main objective is to demonstrate the stability of this solution and the in vivo effects on serum and liver parameters of glucose and lipid metabolism. After 180 days of analysis the COS showed physicochemical stability (pH and specific gravity) when submitted to different temperatures (24 °C and 40 °C), but the chemical stability, presented by the content of CY (%) in the formulation, showed that the degradation rate of CY increased 20x when submitted to a stress condition (40 °C). Male Wistar rats were treated with IOS 1.5 (industrial oral solution 1.5 mg.kg-1) or saline for 21 days and IOS 5.0 or COS (5 mg.kg-1) or placebo by gavage for 23 days. Treatment with IOS 1.5 increased liver cholesterol by 30 % without changing liver and body weight in this group. In contrast, treatment with IOS and COS (5 mg.kg-1) increased final body weight by 8 % compared to placebo. Interestingly, the COS group had increased liver glycogen accompanied by reduced food intake compared to IOS. However, COS and IOS 5.0 did not induce changes in adipose tissue weight, liver weight, as well as serum glucose, triacylglycerol and cholesterol levels. This work provides us with data that prove the stability of the compound solution at room temperature, as well as inducing an increase in the final body weight of rats with no alteration in glucose and lipid metabolism.

Cholesteryl Ester Transfer Protein Inhibitors and Cardiovascular Outcomes: A Systematic Review and Meta-Analysis.

Atherosclerosis is a multi-factorial disease, and low-density lipoprotein cholesterol (LDL-C) is a critical risk factor in developing atherosclerotic cardiovascular disease (ASCVD). Cholesteryl-ester transfer-protein (CETP), synthesized by the liver, regulates LDL-C and high-density lipoprotein cholesterol (HDL-C) through the bidirectional transfer of lipids. The novelty of CETP inhibitors (CETPis) has granted new focus towards increasing HDL-C, besides lowering LDL-C strategies. To date, five CETPis that are projected to improve lipid profiles, torcetrapib, dalcetrapib, evacetrapib, anacetrapib, and obicetrapib, have reached late-stage clinical development for ASCVD risk reduction. Early trials failed to reduce atherosclerotic cardiovascular occurrences. Given the advent of some recent large-scale clinical trials (ACCELERATE, HPS3/TIMI55-REVEAL Collaborative Group), conducting a meta-analysis is essential to investigate CETPis' efficacy. We conducted a thorough search of randomized controlled trials (RCTs) that commenced between 2003 and 2023; CETPi versus placebo studies with a ≥6-month follow-up and defined outcomes were eligible. major adverse cardiovascular events (MACEs), cardiovascular disease (CVD)-related mortality, all-cause mortality. stroke, revascularization, hospitalization due to acute coronary syndrome, myocardial infarction (MI). Nine RCTs revealed that the use of a CETPi significantly reduced CVD-related mortality (RR = 0.89; 95% CI: 0.81-0.98; p = 0.02; I2 = 0%); the same studies also reduced the risk of MI (RR = 0.92; 95% CI: 0.86-0.98; p = 0.01; I2 = 0%), which was primarily attributed to anacetrapib. The use of a CETPi did not reduce the likelihood any other outcomes. Our meta-analysis shows, for the first time, that CETPis are associated with reduced CVD-related mortality and MI.

Early life exposure to vitamin D deficiency impairs molecular mechanisms that regulate liver cholesterol biosynthesis, energy metabolism, inflammation, and detoxification.

Emerging data suggests liver disease may be initiated during development when there is high genome plasticity and the molecular pathways supporting liver function are being developed. Here, we leveraged our Collaborative Cross mouse model of developmental vitamin D deficiency (DVD) to investigate the role of DVD in dysregulating the molecular mechanisms underlying liver disease. We defined the effects on the adult liver transcriptome and metabolome and examined the role of epigenetic dysregulation. Given that the parental origin of the genome (POG) influences response to DVD, we used our established POG model [POG1-(CC011xCC001)F1 and POG2-(CC001xCC011)F1] to identify interindividual differences. We found that DVD altered the adult liver transcriptome, primarily downregulating genes controlling liver development, response to injury/infection (detoxification & inflammation), cholesterol biosynthesis, and energy production. In concordance with these transcriptional changes, we found that DVD decreased liver cell membrane-associated lipids (including cholesterol) and pentose phosphate pathway metabolites. Each POG also exhibited distinct responses. POG1 exhibited almost 2X more differentially expressed genes (DEGs) with effects indicative of increased energy utilization. This included upregulation of lipid and amino acid metabolism genes and increased intermediate lipid and amino acid metabolites, increased energy cofactors, and decreased energy substrates. POG2 exhibited broader downregulation of cholesterol biosynthesis genes with a metabolomics profile indicative of decreased energy utilization. Although DVD primarily caused loss of liver DNA methylation for both POGs, only one epimutation was shared, and POG2 had 6.5X more differentially methylated genes. Differential methylation was detected at DEGs regulating developmental processes such as amino acid transport (POG1) and cell growth & differentiation (e.g., Wnt & cadherin signaling, POG2). These findings implicate a novel role for maternal vitamin D in programming essential offspring liver functions that are dysregulated in liver disease. Importantly, impairment of these processes was not rescued by vitamin D treatment at weaning, suggesting these effects require preventative measures. Substantial differences in POG response to DVD demonstrate that the parental genomic context of exposure determines offspring susceptibility.

Berberine alleviates cholesterol and bile acid metabolism disorders induced by high cholesterol diet in mice

Berberine (BBR) is a traditional Chinese herb with broad antimicrobial activity. Gut microbiota plays an important role in the metabolism of bile acids and cholesterol. Our study investigated the effects of BBR on alleviating cholesterol and bile acid metabolism disorders induced by high cholesterol diet in mice. Adult male C57BL/6J mice fed with high cholesterol diet (HC) containing 1.25 % cholesterol (HC group) or fed with chow diet containing 0.02 % cholesterol (Chow group) served as controls. BBR50 and BBR100 group mice were fed with HC, and oral BBR daily at doses of 50 or 100 mg/kg respectively for 8 weeks. The results showed that BBR could reshape the homeostasis and composition of gut microbiota. The abundance of Clostridium genera was significantly inhibited by BBR, which resulted in a significant reduction of secondary bile acids within the enterohepatic circulation and a significant lower hydrophobic index of bile acids. The absorption of cholesterol in intestine, the deposition of cholesterol in liver and the excretion of cholesterol in biliary tract were significantly inhibited by BBR, which promoted the unsaturation of cholesterol in bile. These findings suggest the potential utility of BBR as a functional food to alleviate the negative effects of high cholesterol diet.

Metabolic Effects of Short-Term High-Fat Intake Vary Depending on Dietary Amino Acid Composition

BackgroundIt is generally accepted that excessive fat intake has undesirable effects on the energy metabolism of our body. Dietary amino acid composition is also critical to the regulation of lipid metabolism. ObjectivesThis study aimed to investigate whether high-fat diets (HFDs) with different amino acid deficiencies lead to different metabolic outcomes. MethodsSix-wk-old male Wistar rats were fed either a control diet (CN; 3.7 kcal/g, 12% calories from fat) or HFDs (5.1 kcal/g, 60% calories from fat) with 7 different amino acid compositions [control or methionine, arginine, histidine, lysine, threonine, or branched-chain amino acids (BCAAs) deficient], for 7 d. Tissue weights and lipid accumulation in the liver, skeletal muscle, and adipose tissue were measured, and serum biochemical parameters were analyzed. ResultsAlthough the food intake of the HFD groups was a little less than that of the CN group, the total calorie intakes were comparable among the groups, except for histidine-deficient and BCAA-deficient groups. In rats fed am HFD with a control amino acid composition (HFCN), dramatic increase in triglyceride (TG) accumulation in the liver and serum LDL cholesterol concentration were observed compared with the CN group. However, when the arginine content in the diet was reduced, liver TG accumulation was completely inhibited, with no apparent effects on serum lipoprotein-cholesterol concentrations. Meanwhile, deficiency of the other amino acids, such as threonine, reversed HFD-induced upregulation of serum LDL cholesterol. ConclusionsIt is observed that although the rats ingested an excessive amount of fat, neither ectopic fat accumulation nor dyslipidemia were always induced at least in the short term; hence, the consequent metabolic change was dependent on the dietary amino acid composition. These findings introduce an important perspective regarding HFD regimens in both scientific and clinical contexts.

Subacute cadmium exposure changes different metabolic functions, leading to type 1 and 2 diabetes mellitus features in female rats.

Cadmium (Cd) is a heavy metal that acts as endocrine disrupting chemical (EDC). Few studies have investigated the effects of Cd exposure on metabolic dysfunctions, such as type 1 and 2 diabetes mellitus (T1DM and T2DM). Thus, we assessed whether subacute Cd exposure at occupational levels causes abnormalities in white adipose tissue (WAT), liver, pancreas, and skeletal muscle. We administered cadmium chloride (CdCl2) (100 ppm in drinking water for 30 days) to female rats and evaluated Cd levels in serum and metabolic organs, morphophysiology, inflammation, oxidative stress, fibrosis, and gene expression. High Cd levels were found in serum, WAT, liver, pancreas, and skeletal muscle. Cd-exposed rats showed low adiposity, dyslipidemia, insulin resistance, systemic inflammation, and oxidative stress compared to controls. Cd exposure reduced adipocyte size, hyperleptinemia, increased cholesterol levels, inflammation, apoptosis and fibrosis in WAT. Cd-exposed rats had increased liver cholesterol levels, insulin receptor beta (IRβ) and peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC1α) expression, karyomegaly, inflammation, and fibrosis. Cd exposure reduced insulin levels and pancreatic islet size and increased inflammation and fibrosis. Cd exposure reduced skeletal muscle fiber diameter and increased IR expression and inflammation. Finally, strong positive correlations were observed between serum, tissue Cd levels, abnormal morphology, tissue inflammation and fibrosis. Thus, these data suggest that subacute Cd exposure impairs WAT, liver, pancreas and skeletal muscle function, leading to T1DM and T2DM features and other complications in female rats.

Dual Deletion of Keap1 and Rbpjκ Genes in Liver Leads to Hepatomegaly and Hypercholesterolemia.

The hepatic deletion of Rbpjκ (RbpjF/F::AlbCre) in the mouse leads to exhibition of the Alagille syndrome phenotype during early postnatal liver development with hyperlipidemia and cholestasis due to attenuated disruption of NOTCH signaling. Given the roles of NRF2 signaling in the regulation of lipid metabolism and bile ductal formation, it was anticipated that these symptoms could be alleviated by enhancing NRF2 signaling in the RbpjF/F::AlbCre mouse by hepatic deletion of Keap1 in compound Keap1F/F::RbpjF/F::AlbCre mice. Unexpectedly, these mice developed higher hepatic and plasma cholesterol levels with more severe cholestatic liver damage during the pre-weaning period than in the RbpjF/F::AlbCre mice. In addition, hypercholesterolemia and hepatic damage were sustained throughout the growth period unlike in the RbpjF/F::AlbCre mouse. These enhanced abnormalities in lipid metabolism appear to be due to NRF2-dependent changes in gene expression related to cholesterol synthetic and subsequent bile acid production pathways. Notably, the hepatic expression of Cyp1A7 and Abcb11 genes involved in bile acid homeostasis was significantly reduced in Keap1F/F::RbpjF/F::AlbCre compared to RbpjF/F::AlbCre mice. The accumulation of liver cholesterol and the weakened capacity for bile excretion during the 3 pre-weaning weeks in the Keap1F/F::RbpjF/F::AlbCre mice may aggravate hepatocellular damage level caused by both excessive cholesterol and residual bile acid toxicity in hepatocytes. These results indicate that a tuned balance of NOTCH and NRF2 signaling is of biological importance for early liver development after birth.

Nano Formulation and Evaluation of Atorvastatin Mouth Dissolving Tablets

Atorvastatin calcium is an anti-hyperlipidemic drug that competitively inhibit HMG-CoA reductase and decrease production of cholesterol in liver. This statin derivative is highly metabolized in gut wall and liver so that it produces only 14% oral bioavailability. In this study we aim to improve the bioavailability of BCS II drug atorvastatin calcium in the formulation of mouth dissolving tablet, which enters the blood stream via facial, jugular artery and vein, by penetrating the lipid membrane when the tablet disintegrates. The Atorvastatin – chitosan nanoparticles was formulated by freeze drying of colloidal solution of nanoparticles. Super disintegrant like croscarmellose sodium, crospovidone, and sodium starch glycolate were added and tested for its dispersion properties. The prepared formulation shows enhanced rate of dissolution and may helpful by avoiding first pass metabolism and to achieve maximum bioavailability. Further the polymeric nano formulation (atorvastatin-chitosan nano particle) allows drug to penetrate much faster and dissolve the drug in saliva. Finally, the prepared formulation is evaluated by pre formulation studies, pre compression evaluation, post compression evaluation and the result of the study revealed that atorvastatin chitosan nanoparticles with size of 406 nm and PDI 0.246 had 118% dissolute in 10 min and shows enhance bioavailability.

Physiologically based pharmacokinetics modeling and transporter proteomics to predict systemic and local liver and muscle disposition of statins.

Statins are used to reduce liver cholesterol levels but also carry a dose-related risk of skeletal muscle toxicity. Concentrations of statins in plasma are often used to assess efficacy and safety, but because statins are substrates of membrane transporters that are present in diverse tissues, local differences in intracellular tissue concentrations cannot be ruled out. Thus, plasma concentration may not be an adequate indicator of efficacy and toxicity. To bridge this gap, we used physiologically based pharmacokinetic (PBPK) modeling to predict intracellular concentrations of statins. Quantitative data on transporter clearance were scaled from invitro to invivo conditions by integrating targeted proteomics and transporter kinetics data. The developed PBPK models, informed by proteomics, suggested that organic anion-transporting polypeptide 2B1 (OATP2B1) and multidrug resistance-associated protein 1 (MRP1) play a pivotal role in the distribution of statins in muscle. Using these PBPK models, we were able to predict the impact of alterations in transporter function due to genotype or drug-drug interactions on statin systemic concentrations and exposure in liver and muscle. These results underscore the potential of proteomics-guided PBPK modeling to scale transporter clearance from invitro data to real-world implications. It is important to evaluate the role of drug transporters when predicting tissue exposure associated with on- and off-target effects.

Pu-erh tea partly improved liver cholesterol metabolism disorders in colitis mice via the gut-liver axis

Pu-erh tea partly improved liver cholesterol metabolism disorders in colitis mice <em>via</em> the gut-liver axis

Abstract 2799: Intestinal Akkermansia muciniphila complements the therapeutic efficacy of PD1 therapy by reshaping the immunosuppressive microenvironment in nonalcoholic fatty liver disease (NAFLD)-induced hepatocellular carcinoma

Abstract Objectives: Nonalcoholic fatty liver disease (NAFLD)-induced hepatocellular carcinoma (HCC) is an emerging malignancy in the developed world. However, immune checkpoint inhibitors (ICIs) are ineffective for patients with this disease. Advances in research have shown that gut-liver communication contributes to NAFLD progression. Therefore, identifying a key gut microbiota for predicting ICIs response and an effective combinatorial strategy is urgently needed. Design: 16S rRNA sequencing was employed to identify key gut microbiota crucial in developing the mouse NAFLD-induced HCC model. The in vivo roles of Akkermansia muciniphila (Akk) were evaluated by immunofluorescence staining, qPCR, mass spectrometry analysis, ELISA assay, and single-cell RNA sequencing (scRNA-seq) coupled with immune profiling analysis. The clinical and therapeutic value of Akk alone in combination with PD1 treatment was investigated. Results: Akk was decreased from healthy to HCC tissues during HCC tumor development, and daily administration of Akk not only ameliorates liver steatosis and cholesterol biosynthesis but could effectively attenuate the development of NAFLD-induced HCC. Akk repairs the intestinal lining, with a concurrent decrease in the serum concentration of lipopolysaccharide (LPS) and bile acid metabolites. Using scRNA-seq coupled with immune profiling analyses, we found that Akk suppressed the populations of immunosuppressive cells, including monocytic myeloid-derived suppressor cells (m-MDSCs) and M2 macrophages, by suppressing the differentiation of these cells from monocytes, which leads to T cell proliferation and activation. Consistent with this finding, Akk administration, combined with PD1 treatment, exerted a maximal growth-suppressive effect, accompanied by increased infiltration and activation of T cells. Clinically, the fecal level of Akk in HCC patients was decreased in NAFLD-induced HCC patients and serves as a potential biomarker for predicting PD1 response in HCC patients. Conclusions: Akk regulates the immune tumor environment by regulating two key immune suppressive cells via regulation of the LPS flux to develop NALFD-induced HCC. This finding provides a solid therapeutic basis for a rational combination with PD1 for the treatment of this deadly disease. Interestingly, Akk may serve as a predictive biomarker for PD1 response in HCC patients. Citation Format: Xueqian Wu, Fan Ying, Katherine Po Sin Chung, Carmen Oi Ning Leung, Terence Kin Wah Lee. Intestinal Akkermansia muciniphila complements the therapeutic efficacy of PD1 therapy by reshaping the immunosuppressive microenvironment in nonalcoholic fatty liver disease (NAFLD)-induced hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2799.

Therapeutic potential of caffeic acid and coumarin in modifying aluminum-induced hepatic injury in Wistar rats

Antioxidants and phenolic compounds have always been tested as chemoprotective agents to regulate disease progression related to oxidative stress. Through this study, we investigate the possible corrective effects of caffeic acid and coumarin against aluminum- (Al-) induced stress response and liver damage in rats. Male Wistar rats were divided into four groups (10 rats per group), cotreated with caffeic acid (30 mg kg-1) and coumarin (5 mg kg-1) and exposed to aluminum (60 mg kg-1) for 45 days. Al induced significant alterations in body and liver weight over time. It also led to a marked increase in plasma levels of transaminases (aspartate aminotransferase and alanine aminotransferase), alkaline phosphatase, and lactate dehydrogenase. Additionally, there was a substantial change in the lipid profile, characterized by elevated levels of triglycerides and total cholesterol in both serum and liver. Furthermore, the LDL-c level and the HDL-c/total cholesterol ratio in serum were also affected. In contrast, the levels of HDL-c in serum and phospholipids in liver tissues were reduced. The results of this study also showed increased levels of TBARS, protein carbonyls, nitrates/nitrites, and TNF-α. Conversely, the activity of antioxidant enzymes (catalase, GP-x, and SOD) was reduced. These altered parameters were restored after treating rats with caffeic acid and coumarin. Our study strongly suggests that caffeic acid and coumarin possess antioxidant properties and hepatoprotective capacity by attenuating aluminum toxicity-induced liver damage.