Bile Acid Binding Research Articles

Exploiting the bile acid binding protein as transporter of a Cholic Acid/Mirin bioconjugate for potential applications in liver cancer therapy.

Bioconjugation is one of the most promising strategies to improve drug delivery, especially in cancer therapy. Biomolecules such as bile acids (BAs) have been intensively explored as carriers, due to their peculiar physicochemical properties and biocompatibility. BAs trafficking is regulated by intracellular lipid-binding proteins and their transport in the liver can be studied using chicken liver Bile Acid-Binding Proteins (cL-BABPs) as a reference model. Therefore, we conceived the idea of developing a BA-conjugate with Mirin, an exonuclease inhibitor of Mre11 endowed with different anticancer activities, to direct its transport to the liver. Following computational analysis of various BAs in complex with cL-BABP, we identified cholic acid (CA) as the most promising candidate as carrier, leading to the synthesis of a novel bioconjugate named CA-M11. As predicted by computational data and confirmed by X-ray crystallographic studies, CA-M11 was able to accommodate into the binding pocket of BABP. Hence, it can enter BAs trafficking in the hepatic compartment and here release Mirin. The effect of CA-M11, evaluated in combination with varying concentrations of Doxorubicin on HepG2 cell line, demonstrated a significant increase in cell mortality compared to the use of the cytotoxic drug or Mirin alone, thus highlighting chemo-sensitizing properties. The promising results regarding plasma stability for CA-M11 validate its potential as a valuable agent or adjuvant for hepatic cancer therapy.

In Vitro Antioxidant and Hypocholesterolemic Potency of Melon (Cucumis melo L.) Seed Protein Hydrolysate

Protein hydrolysates have been reported to possess numerous bioactivities. However, research on melon (Cucumis melo L.) seed protein (MSP) hydrolysate is limited. This study aimed to analyze the antioxidant and hypocholesterolemic properties of MSP hydrolysate. Protein from melon seed was obtained by conventional alkaline extraction-isoelectric precipitation method. Enzymatic hydrolysis of MSP was carried out using three different proteases: pepsin, thermolysin, and trypsin, with the enzyme-to-substrate (E/S) ratio of 1:50 (w/w). The results showed that all hydrolysates exhibited antioxidant and hypocholesterolemic activity. The thermolysin-digested hydrolysate had significantly greater (p < 0.05) radical scavenging properties, while trypsin produced the highest (p < 0.05) metal ion chelating activity. At 2 mg/mL, thermolysin-derived MSP hydrolysate showed no significant difference (p > 0.05) in HMGR inhibition activity compared to pravastatin. Additionally, the thermolysin hydrolysate had significantly higher (p < 0.05) bile acid binding ability than other hydrolysates. Overall, the MSP hydrolysate produced by thermolysin exhibited stronger antioxidant and hypocholesterolemic activities when compared to those produced by pepsin and trypsin, suggesting its potential effectiveness in nutraceutical applications.

Metabolic immaturity and breastmilk bile acid metabolites are central determinants of heightened newborn vulnerability to norovirus diarrhea

The pathogenic outcome of enteric virus infections is governed by a complex interplay between the virus, intestinal microbiota, and host immune factors, with metabolites serving as a key mediator. Noroviruses bind bile acid metabolites, which are produced by the host and then modified by commensal bacteria. Paradoxically, bile acids can have both proviral and antiviral roles during norovirus infections. Working in an infant mouse model of norovirus infection, we demonstrate that microbiota and their bile acid metabolites protect from norovirus diarrhea, whereas host bile acids promote disease. We also find that maternal bile acid metabolism determines the susceptibility of newborn mice to norovirus diarrhea during breastfeeding. Finally, targeting maternal and neonatal bile acid metabolism can protect newborn mice from norovirus disease. In summary, neonatal metabolic immaturity and breastmilk bile acids are central determinants of heightened newborn vulnerability to norovirus disease.

Suppressive Effect of Coffee Leaves on Lipid Digestion and Absorption In Vitro.

Coffee leaves are a major source of bioactive components and are used as ethnomedicine. However, despite their traditional medicinal use, information about their effects on antihyperlipidemia remains limited. The aims of this study were to evaluate the main components of leaf extracts from Arabica and Robusta coffees and to examine the potential of these coffee leaves in reducing lipid digestion and absorption in vitro. Coffee leaf extracts from Arabica coffee contain a high amount of caffeine, whereas extracts from Robusta coffee contain high amounts of chlorogenic acid (CGA) and caffeine. Additionally, leaf extracts from Arabica and Robusta coffee demonstrated the inhibition of pancreatic lipase, decreased micellar cholesterol solubility, and reduced bile acid binding. Furthermore, these extracts resulted in a reduction in cholesterol uptake in Caco-2 cells. Molecular docking experiments supported this discovery, showing CGA and caffeine binding to Niemann-Pick C1-like 1 (NPC1L1), a key protein in cholesterol absorption. The results indicated that CGA and caffeine can competitively bind to NPC1L1 at the cholesterol binding pocket, reducing its cholesterol binding rate. These findings suggest that coffee leaves might help suppress lipid absorption and digestion, highlighting their potential use in preventing and treating hyperlipidemia.

Fabrication of resveratrol-loaded soy peptide nanogels with transglutaminase and in vitro gastrointestinal delivery and cholesterol-lowering effect

To improve the intestinal delivery of resveratrol, soy protein peptides (SPP) and transglutaminase (TGase) were used to prepare nanogels (NGs). The results demonstrated that, compared with native soy protein isolate (SPI) NG, SPP-NG showed a lower encapsulation efficiency (EE, 70.26%), a higher release efficiency (RE, 95.83%), and a smaller particle size (194.43 nm). However, the addition of TGase increased the EE and particle size to 74.37% and 217.73 nm, respectively. The RE of SPP-TGNG was also maintained at as high as 93.28%. Interestingly, SPP-TGNG exhibited optimal bile acid–binding capacity and cholesterol-lowering digests. The differences in delivery effects and morphology were attributed to structural changes in NGs. The unfolding and exposure of hydrophobic residues and weakened hydrogen bonds by hydrolysis resulted in a disordered and loose network structure, but it was beneficial to the production of cholesterol-lowering peptides and amino acids after digestion. The presence of TGase strengthened the crosslinking network by increasing β-sheet contents, hydrogen bonds, hydrophobic interactions, and isopeptide bonds. The alteration in multilevel structure would further affect the digestive products. The exposure of more hydrophobic residues and isopeptide crosslinking may be the intrinsic reason for the production of more hypocholesterolemic peptides and amino acids. This study indicated that the combination of hydrolysis and TGase treatment enhanced the delivery efficiency and bioactive properties of peptide-based NGs to resveratrol and revealed the internal mechanism between the structural changes and delivery efficiency. This work offers a theoretical foundation and technical suggestions for the production of peptide-based cargo systems and hydrophobic active substances.

The GLP-1 Receptor Agonist Liraglutide Decreases Primary Bile Acids and Serotonin in the Colon Independently of Feeding in Mice.

Liraglutide, a glucagon-like peptide 1 analog used to treat type 2 diabetes and obesity, is a potential new treatment modality for bile acid (BA) diarrhea. Here, we show that administration of liraglutide significantly decreased total BAs, especially the primary BAs, including cholic acid, chenodeoxycholic acid, taurocholic acid, taurochenodeoxycholic acid, glycocholic acid, and β-muricholic acid, in the liver and feces. In addition, liraglutide significantly decreased tryptophan metabolites, including L-tryptophan, serotonin, 5-hydroxy indole-3-acetic acid, L-kynurenine, and xanthurenic acid, in the colon, whereas it significantly increased indole-3-propionic acid. Moreover, the administration of liraglutide remarkably decreased the expression of apical sodium-dependent bile acid transporter, which mediates BA uptake across the apical brush border member in the ileum, ileal BA binding protein, and fibroblast growth factor 15 in association with decreased expression of the BA-activated nuclear receptor farnesoid X receptor and the heteromeric organic solute transporter Ostα/β, which induces BA excretion, in the ileum. Liraglutide acutely decreased body weight and blood glucose levels in association with decreases in plasma insulin and serotonin levels in food-deprived mice. These findings suggest the potential of liraglutide as a novel inhibitor of primary BAs and serotonin in the colon.

Black Goji Berry (Lycium ruthenicum) Juice Fermented with Lactobacillus rhamnosus GG Enhances Inhibitory Activity against Dipeptidyl Peptidase-IV and Key Steps of Lipid Digestion and Absorption.

With the global increase in hyperglycemia and hyperlipidemia, there is an urgent need to explore dietary interventions targeting the inhibition of dipeptidyl peptidase-IV (DPP-IV) and lipid digestion and absorption. This study investigated how Lactobacillus rhamnosus GG (LGG) affects various aspects of black goji berry (BGB) (Lycium ruthenicum Murr.) juice, including changes in physicochemical and functional properties, as well as microbiological and sensory attributes. Throughout the fermentation process with 2.5-10% (w/v) BGB, significantly improved probiotic viability, lactic acid production, and decreased sugar content. While total flavonoids increase, anthocyanins decrease, with no discernible change in antioxidant activities. Metabolite profiling reveals elevated phenolic compounds post-fermentation. Regarding the inhibition of lipid digestion and absorption, fermented BGB exhibits improved bile acid binding, and disrupted cholesterol micellization by approximately threefold compared to non-fermented BGB, while also increasing pancreatic lipase inhibitory activity. Furthermore, a decrease in cholesterol uptake was observed in Caco-2 cells treated with fermented BGB (0.5 mg/mL), with a maximum reduction of 16.94%. Fermented BGB also shows more potent DPP-IV inhibition. Sensory attributes are significantly improved in fermented BGB samples. These findings highlight the potential of BGB as a bioactive resource and a promising non-dairy carrier for LGG, enhancing its anti-hyperglycemic and anti-hyperlipidemic properties.

Colesevelam for Lenalidomide Associated Diarrhea in Patients with Multiple Myeloma.

Lenalidomide maintenance is associated with a significantly improved progression-free in patients with newly diagnosed multiple myeloma. Maintenance with lenalidomide is generally well tolerated; however, lenalidomide associated diarrhea is a common side effect and bile acid malabsorption has been suggested as an underlying mechanism. We conducted a single arm phase 2 trial of colesevelam, a bile acid binder, for lenalidomide-associated diarrhea in multiple myeloma. Patients were treated with colesevelam daily starting at 1250 mg (2 tablets 625 mg) for 12 weeks. The trial included 25 patients, 1 patient with grade 3 diarrhea, 14 with grade 2, and 10 with grade 1 diarrhea. All patients were on treatment with single agent lenalidomide maintenance and no patient progressed during the trial. Colesevelam treatment was highly effective for treatment of lenalidomide-associated diarrhea; 22 (88%) of the 25 patients responded where 17 patients (68%) had complete resolution of diarrhea, and 5 patients (20%) had improvement by 1 grade of diarrhea. The responses to colesevelam were seen within the first two weeks of treatment. These findings support the conclusion that lenalidomide-associated diarrhea is driven by bile acid malabsorption. Five patients reported mild gastrointestinal side effects including constipation. Importantly, the pharmacokinetics of lenalidomide were not affected by concomitant colesevelam treatment. The stool microbiome composition was not significantly different before and after colesevelam treatment. Patients reported improved diarrhea, fewer gastrointestinal symptoms, and less interference with their daily life after starting colesevelam. In summary, colesevelam was safe and highly effective for treatment of lenalidomide-associated diarrhea in multiple myeloma and does not reduce the clinical effect of lenalidomide.

#3067 Comparison of the removal of bile acids between hemoperfusion and new extracorporeal albumin detoxification device (Albunique) in critical care

Abstract Background and Aims In subjects with multiorgan failure and in need for CRRT, new onset of jaundice is associated with a significant mortality. Recent research suggests that elevated serum bile acids contribute to severe immune dysfunction, hepatocellular apoptosis, vasodilation, tubular necrosis and thereby are important drivers of multi-organ failure in new onset of liver dysfunction. Extracorporeal Albumin Detoxification and Dialysis (ECAD) and Hemoperfusion have been used to sustain or recover organ function. Recently, Cytosorb, a hemoperfusion device developed for the removal of proinflammatory molecules in cytokine storm showed a reduction of bilirubin and bile acids in vitro. The aim of this study was to compare the reduction of small albumin bound bile acids with the current standard (ECAD), which was developed not to affect the plasma proteome but effectively remove toxins. Method After obtaining consent, plasma samples were obtained from the extracorporeal circuit of subjects on CRRT due to acute on chronic liver failure (ACLF) and bile acids above 50 µmol/l before and after a day of treatment from the venous and arterial ports and the total removed bile acids were calculated by Area under the curve calculation. The measurements were done during Cytosorb treatments versus during ECAD using a new Albunique Device based on a new recycling technology. Results Despite a high initial reduction rate the capacity of Cytosorb decreased and the total amount of extracorporeal removal was 1720+1189 µmol whereas Albunique removed 7111+2556 µmol, more than 4 times as much and was not saturated after 24 hours. Conclusion Although Cytosorb may prove a role in ACLF with hyperinflammation in the future, hemoperfusion appears to loose efficacy for toxic, albumin bound small molecules and therapies based on diffusion appear to be more cost effective and less additional effects with an unclear impact on the course of ACLF.

Bile acid malabsorption investigated by selenium-75-homocholic acid taurine (75SeHCAT) scans, a retrospective single-centre experience.

Bile acid malabsorption (BAM) is a common entity in patients experiencing chronic watery diarrhea. However, literature suggests that BAM is underdiagnosed and undertreated. In many countries 75SeHCAT is the gold standard for diagnosing BAM (1-5). The aim of this retrospective study was to investigate the frequency of BAM using 75SeHCAT scintigraphy and the response to treatment with bile acid sequestrants. We retrospectively evaluated the clinical data of 420 patients who had a 75SeHCAT scan from January 2016 to January 2023. Electronic medical records were investigated for risk factors of BAM. BAM was defined as a 75SeHCAT retention of less than 15%. Response to bile acid sequestrants was evaluated based on outpatient clinical reports. Median age at the time of 75SeHCAT test was 47 years (range 14-87). Out of 420 patients, 192 (46%) had a positive result for BAM. Cholecystectomy and ileal resection were significant predictors for BAM (p<0.001). BAM type 1 is statistically more likely to be more severe compared to type 2 (p<0.0001) and compared to type 3 (p=0.0021). In patients who fulfilled Rome IV criteria for IBS-D or functional diarrhea, 75 out of 121 (38%) tested positive for BAM. Overall, treatment with bile acid binders was effective in 76%. This retrospective study of patients who underwent SeHCAT scanning confirms that about 1/3 patients presenting with therapy-resistant chronic diarrhea in secondary care suffer from BAM. Including the test in clinical management protocols of chronic diarrhea allows early and quantitative diagnosis and treatment of BAM.

Cereal-based fermented synbiotic instant powders: a dessert practice

Objective: This study was aimed to develop synbiotic and probiotic instant powder mixtures that can be used in food products to meet the increasing interest of consumers in functional foods. Materials and Methods Cereal flours (oat, rice, wheat), sesame, cranberry, chestnut and milled germinated grains (lentil, mung bean) were blended and autoclaved then subjected to fermentation process using the Lactobacillus plantarum strain. A part of the porridge was conventionally dried (FPP) and skim milk powder+fructooligosaccharide+inulin mixture as prebiotic sources were added to the other part and subjected to freeze drying (FSP). The viability of probiotics in the gastrointestinal environment and the Angiotensin-converting enzyme inhibition and bile-acid binding capacities of the powders after in vitro digestion were analyzed. The sensory acceptability of the desserts was evaluated. Results: FSP contained a higher number of viable cells than FPP after in vitro digestion. Relative bile-acid binding and angiotensin-converting enzyme inhibition capacities of samples were confirmed as their cholesterol-lowering and blood pressure-regulating potential. The panelists rated the dessert samples enriched with 5-15% FPP and FSP as 'liked'. Conclusion: The integration of these powders into a variety of food products will provide consumers with healthier dietary choices that support their overall health goals.

β-Glucan content and in vitro bile-acid binding capacity of Agaricus bisporus and Pleurotus spp.

The cholesterol lowering properties of oats and barley, attributed to their high β-glucan content, are well established, but it remains unclear whether mushrooms, also rich in β-glucan, exhibit a similar functionality. We aimed to quantify the β-glucan content of commonly consumed Australian mushrooms and evaluated their bile acid binding capacity, the primary cholesterol lowering mechanism of β-glucan. Raw, boiled and fried Australian grown Agaricus bisporus (button, cup, flat and brown mushrooms) and Pleurotus spp. (shimeji and oyster) along with oats were freeze-dried and the β-glucan content was determined. The bile acid binding capacity of these samples was assessed using an in vitro digestion assay. The β-glucan content of freeze-dried raw A. bisporus mushrooms (4.5-8.1 g per 100 g) was similar to that of oats (7.6 g per 100 g, all p > 0.05), whereas Pleurotus mushrooms contained ∼5 times more β-glucan (32.5-37.4 g, p < 0.05). Boiling increased the β-glucan content of oyster, button, flat and brown mushrooms by 3-7% (p < 0.05), but did not affect the β-glucan content of shimeji or cup mushrooms. Frying had no effect on any mushroom type. The bile acid binding capacity of A. bisporus mushrooms (29-36%) was equivalent to that of raw oats (36%, p > 0.05), whereas the bile acid binding capacity of oyster mushrooms (22%) was lower than that of oats (p < 0.05). Both boiling and frying increased the bile acid binding capacity. The cholesterol lowering effects of A. bisporus mushrooms and the acceptability of consumption at the required levels need to be confirmed by clinical trials.

Oxidative Stress: Use of Alternative Therapeutic Agents

Kalonji seeds are a powerhouse of crude fibers, amino acids, iron, sodium, calcium, and potassium. Kalonji is also rich in vitamins that include – Vitamin A, Vitamin B, Vitamin B12, Niacin, and Vitamin C. Kalonji oil is much better than other oil as it contains essential fatty acids, vitamins, and minerals. It comprised approximately 17% protein, 26% carbohydrates, and 57% plant fats and oils. Apart from drugs various substances and life style changes are being advised to correct oxidative stress burden in human body. Analysis of the lipid content in the oxidatively modified LDL from hyper lipidemic patients revealed that the level of lysophosphatidylcholine was greatly elevated, and the high level of the lysolipid was shown to impair the endothelium-dependent relaxation of the blood vessels. In alllopathy blood lipid levels are normalized by statins, fibrates, niacin and bile acid binding resins (BABRs). All of these allopathy related drugs have low compliance ranges from mild compliance due to metalic taste of BABRs to severe side effects like rhabdomyolysis by fibrates and statins. Cardiovascular diseases are leading cause of death in western and eastern countries of the world. Hyperlipidemia is one of the strong risk fractions for heart diseases. To evaluate hypolipidemic drugs efficacy, the study was conducted at Ghurki trust teaching hospital, Lahore Pakistan from January 2020 to August 2020. Ninty hyperlipidemic patients were selected from cardiology and medical wards of the hospital. They were divided in three groups, one at placebo therapy, another on Kalonji and third one on Vitamin B3. After one and half month, significant changes (p value ranging from &lt;0.05 to &lt;0.001) were observed in their LDL and HDL-cholesterol. Conclusion of the study was to recommend use of herbal medicine and vitamin B3 for prevention of any heart diseases with good patient compliance. KEY WORDS: kalonji, LDL cholesterol, HDL cholesterol, blood vessels.

Resveratrol ameliorates intestinal lipid metabolism through the PPAR signaling pathway in high-fat diet-fed red tilapia (Oreochromis niloticus)

Feeding high-fat (HF) diets has been shown to cause hepatic and intestinal impairment in fish species, but the mode of action, especially the pathways involved in the intestine, has not been determined yet. In this study, the effects of resveratrol (RES) supplementation on the intestinal structure, microbial flora, and fat metabolism in red tilapia (Oreochromis niloticus) were determined. The results showed RES maintained the structural integrity of the intestine and significantly increased the number of goblet cells in the midgut. RES significantly induced interferon (IL)-1β, IL-6, IL-10, and tumor necrosis factor (TNF)-α, serumal and fecal trimetlylamine oxide (TMAO) and lipopolysaccharides (LPS), intestinal acetic acid levels. However, the concentrations of bound bile acids increased in HF-fed red tilapia. Atp5fa1 and Pafah1b3 significantly increased, Pmt and Acss2 significantly decreased, respectively, with RES supplementation, which was alleviated and retained at the same level in the selisistat (EX527) group. While for transcriptome and proteomics results, RES was found to promote fatty acid β-oxidation and arachidonic acid metabolism associated with the peroxisome proliferator-activated receptor (PPAR) signaling pathway. The next validation experiment showed some genes related to apoptosis and fatty acid metabolism pathways were altered by RES supplementation. Namely, sn6, loc100702698, new_14481, and prkaa1 were upregulated, while ffrs1, ap3s1, and loc100705861 were downregulated. RES significantly increased Planctomycetes and Verrucomicrobia while decreased Moonvirus, Citrobacter, and Pseudomonas. Akkermansia and Fusobacterium significantly increased and Aeromonas significantly decreased. Thus, unsaturated fatty acid biosynthesis significantly increased and carbohydrate/energy metabolism decreased. To conclude, RES enabled the body to complete fatty acid β-oxidation and arachidonic acid metabolism, whereas the addition of inhibitors increased the expression of the phagosome transcriptome and reduced fatty acid β-oxidative metabolism.

Enhancing Phytochemical Compounds, Functional Properties, and Volatile Flavor Profiles of Pomelo (Citrus grandis (L.) Osbeck) Juices from Different Cultivars through Fermentation with Lacticaseibacillus paracasei.

The current study aimed to explore the effects of fermenting five different pomelo cultivars using Lacticaseibacillus paracasei on various physicochemical, phytochemical, and organoleptic attributes. Fermentation led to an increase in viable lactic acid bacteria count (8.80-9.28 log cfu/mL), organic acids, total polyphenols, and flavonoids, resulting in improved antioxidant activity, bile acid binding, cholesterol micellization disruption, and inhibition of pancreatic lipase activity. Additionally, some cultivars displayed higher levels of naringin, naringenin, and hesperetin after fermentation. The levels of volatile compounds were elevated after fermentation. The bitterness and overall acceptability scores were improved in the fermented samples of the Kao Numpueng cultivar. The principal component analysis (PCA) revealed that the Tubtim Siam cultivar demonstrated the highest functionality and health-related benefits among all fermented pomelos. Overall, the study suggests that pomelo exhibits potential as a valuable resource for creating a dairy-free probiotic drink enriched with bioactive phytochemical compounds and beneficial functional attributes.

Characterization and Bile Acid Binding Capacity of Dietary Fiber Obtained from Three Different Amaranth Products.

Amaranth is a dicotyledonous plant, now considered a health-promoting food. It has been rediscovered by the worldwide food industry, which is increasingly becoming aware of the many uses and benefits provided by amaranth in various food preparations. Amaranth dietary fibers, soluble and insoluble fractions, obtained from flour, protein isolate, and beverage were physicochemically characterized and their potential bile acid binding capacity was evaluated. Primary bile acids binding to fiber might contribute to a hypocholesterolemic effect, while the binding of secondary bile acids could minimize the cytotoxic effect that these metabolites exert on the colon. Amaranth fiber fractions were capable of sequestering cholate, taurocholate, deoxycholate, and bovine bile, with a percentage depending not only on the origin and the type of amaranth fiber evaluated but also on the bile acid studied. Flour fiber and the protein isolate insoluble fractions were the most efficient for binding bile and bile acids with uptake values between 29 and 100% relative to cholestyramine. Moreover, deoxycholate, a hydrophobic secondary bile acid, was the most captured by all the fractions, reaching 100% uptake with total and insoluble fibers of the three amaranth products. These results would suggest that the main effect through which amaranth fiber binds bile acids corresponds to an adsorptive effect mediated by hydrophobic interactions.

A systematical strategy for quality markers screening of different methods processing Platycodonis radix based on phytochemical analysis and the impact on Chronic Obstructive Pulmonary Disease

Ethnopharmacological relevanceBaihezhijiegeng is a processed product of Platycodonis radix, and it's effective in the treatment of Chronic Obstructive Pulmonary Disease (COPD). However, the specific mechanism of action has not been reported in the literature. Aim of the studyWe attempted to evaluate the phytochemical composition and pharmaco-dynamics of Platycodon grandiflorum (PG) and BJ to clarify the mechanism behind the expectorant effect of BJ. Materials and methodsWe integrated the ultra-high-performance liquid chromatography-linear trap quadrupole orbitrap velos mass spectrometry (UPLC-LTQ Orbitrap MS/MS) and the ultra-performance liquid chromatography quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) methods to identify the chemical constituents of PG and BJ. Moreover, correlation and multivariate statistical analyses were utilized to seek the candidate quality markers of PG and BJ. Analysis of effective herbal chemical components using UPLC-Q-TOF-MS/MS and retrieval of COPD disease targets from OMIM, TTD, GeneCard databases. Protein-protein interaction (PPI) and topology analyses were performed using the String database and Cytoscape 3.7.2 software; gene ontology (GO) functional enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed using the Metescape platform on common targets. Moreover, we used molecular docking to predict the potential mechanism of quality markers for developing anti-COPD activity. Simultaneously, the model of COPD was established by exposing the animals to cigarette smoke combined with a tracheal drip injection of lipopolysaccharide (LPS). Using the ELISA method, quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting (WB) to determine tumor-necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and matrix metalloproteinase (MMP)9 levels in serum and IL-4, IL-10, IFN-γ levels, epidermal growth factor receptor (EGFR) and MUC5AC expression in lung tissue of COPD rats to explore the therapeutic effects of PG and BJ on the COPD rat model. ResultsThe chemical identification of JG and PG extracts using UPLC-Q-TOF-MS/MS and UPLC-LTQ Orbitrap MS/MS showed 71 compounds, including 47 saponins, 16 phenolic acids, four flavonoids, and four other components. The multivariate statistical analysis showed that seven quality markers were screened. Network pharmacology results showed a role in biological processes such as cellular response to hydrogen peroxide, positive regulation of pri-miRNA transcription from RNA polymerase II promoter, molecular functions such as oxidoreductase activity, acting on NAD(P)H, quinone or similar compound as acceptor, bile acid binding and other molecular functions. In COPD rats, histopathological findings depicted that BJ administration could effectively inhibit inflammatory cell infiltration and mucus hypersecretion, and improve the lung pathological status in rats with COPD. Moreover, BJ could significantly decrease TNF-α, IL-1β, IL-6, and matrix metalloproteinase (MMP)9 levels in the serum and interferon (IFN)-γ levels in lung tissues of rats with COPD (p < 0.01), and significantly increase IL-4 and IL-10 levels in their lung tissues (p < 0.01), suggesting its inhibition of the inflammatory response in vivo. Additionally, EGFR and MUC5AC were reduced in the lung tissues of rats with COPD and airway mucus hypersecretion in rats with COPD. ConclusionThis study revealed the material basis of PG and BJ for anti-COPD activity and discovered the quality markers of PG and BJ which could affect the anti-COPD activity. The therapeutic effects of BJ may be attributed to the regulation of the inflammatory mediators and mediation of the EGFR/MUC5AC pathway in rats with COPD.

SAT018 Investigate If Sex Differences In Microbiota Is Sufficient To Promote Bile Acid-Mediated Hepatocellular Carcinoma In Mice

Abstract Disclosure: A.E. Dean: None. S. Anakk: None. Liver cancer is one of the leading causes of cancer-related deaths with men having a higher prevalence than women. It is becoming clearer that changes in bile acid levels are seen in patients diagnosed with liver cancer. Although initially made in the liver, these primary bile acids can also be modified by bacteria in the intestines to form secondary bile acids. The secondary bile acids are more hydrophobic and cytotoxic and when in excess, can promote tumorigenesis. We, therefore, examined if microbial differences between males and females explain sex-dependent HCC development when bile acid metabolism is altered. In this study, we used a mouse model of bile acid excess and HCC, where the bile acid-related nuclear receptors, farnesoid X receptor (Fxr) and small heterodimer partner (Shp), are knocked out. This double knockout (DKO) model mimics the cancer incidence observed in humans, where male mice are more susceptible to spontaneous tumorigenesis than females at 12 months of age. The circulating bile acid profiles of the male DKO mice are more hydrophobic and have a greater proportion of secondary bile acids than the females. Since we observed changes in the bile acid composition, we next investigated if fecal and cecal microbial profiles of male and female DKO mice differed, along with their wild-type counterparts. 16s rRNA full-length sequencing was performed on fecal and cecal samples from 12-month-old mice, and analyses are currently ongoing. We expect that the microbes identified in the male DKO mice will reflect their bile acid compositions, as well as spontaneous tumorigenesis. Additionally, we are analyzing the 16s rRNA gene from mice treated with cholestyramine (CHR), a bile acid binding resin, that exhibit decreased tumorigenesis at 12 months of age. In this manner, we can correlate microbiota changes to the bile acid composition, as well as the extent of tumor burden. Thus, this study will be able to integrate how sex differences in liver cancer incidence can be secondary to microbial modification of bile acids. Presentation: Saturday, June 17, 2023

Maximizing hypocholesterolemic peptides from an olive byproduct by enzymatic hydrolysis

Novel food proteases (Maxipro A, Maxipro B, Maxipro C, and Maxipro D) with broad specificity and different effective pH range (from 2.5 to 9.0) were evaluated to produce hypocholesterolemic peptides from olive seeds proteins. Box-Behnken experimental design was employed to evaluate the effect of the hydrolysis time, pH, temperature, and enzyme:substrate ratio on the amount of released peptides. The production of peptides, under optimal conditions, was higher with Maxipro C and lower with Maxipro B. The capability of extracts to reduce the absorption of exogenous cholesterol and the production of endogenous cholesterol was also evaluated. All hydrolysates showed a higher capacity to bind bile acids and inhibit cholesterol esterase enzyme than the obtained with usual Alcalase enzyme. Proteomic analysis using high resolution RP-HPLC-ESI-QTOF with database searching and de novo identification was employed to identify peptides in hydrolysates. Some sequences with hypocholesterolemic features were detected among the 45 peptides identified in Maxipro A hydrolysate, the 56 peptides in that of Maxipro C, and the 50 in the Maxipro B one. This work demonstrates that Maxipro A enzyme can hydrolyze olive seeds proteins, under acidic pH, releasing peptides with multifunctional and higher hypocholesterolemic capacity than the obtained with Alcalase, under alkaline conditions.

Effect of Oat β-Glucan on the Structure and Properties of Soybean Protein Isolate During Maillard Reaction.

Maillard reaction (MR) with oat β-glucan changed the structure of soybean protein isolate (SPI), further leading to the enhancement of its functional properties. SPI was unfolded by MR, and the SPI conjugates with high molecular weight were identified. The water solubility of SPI was improved by cross-linking with hydrophilic β-glucan, while the hydrophobicity also increased along with the unfolding of the SPI. Cross-linking with β-glucan elevated the viscosity of SPI, thus enhancing viscosity-related physiological activities, including bile acid binding ability, fat binding capacity, and hypoglycemic activity, and the functional properties increased as the βG content involved in MR increased.