Cholesterol Degradation Research Articles

SNPs in cytochromes P450 catalyzing cholesterol degradation in brain are associated with Parkinson's disease.

Besides being an essential structural component of plasma membranes and the precursor of many functional compounds and signaling molecules, cholesterol was also proposed to play a role in the etiology and/or manifestation of Parkinson's disease (PD). However, so far systematic investigations on the role of cholesterol and its metabolites present in the brain for the etiology of PD are missing. Here, we investigate for the first time the association of PD with SNPs in the genes of four cytochromes P450 (P450), CYP46A1, CYP39A1, CYP27A1 and CYP7B1, which are critical for the degradation of cholesterol in the brain. Analyzing 1,349 individuals from the PPMI data base, we found 24 SNPs in these four genes, which are significantly over- or under-represented in patients suffering from idiopathic PD (IPD). Studying each of the 362 IPD patients individually, we found that most patients (45%) showed only one associated SNP in one of the four P450 genes, while 31% displayed two associated SNPs and 18% three associated SNPs. The occurrence of some associated SNPs is in the same order of magnitude as SNPs in the GBA (beta-glucocerebrosidase) and thus might reflect a genetic predisposition for PD. As all 24 SNPs were located in introns and 3' untranslated regions, we evaluated the prospective regulatory impact of the surrounding genomic regions by using transcriptome and epigenome data from the Foundational Data Initiative for Parkinson Disease (FOUNDIN-PD). FOUNDIN-PD provides gene expression, open chromatin and DNA methylation data in a cohort of 89 induced pluripotent stem cell (iPSC) lines differentiated to dopaminergic (DA) neurons derived from people in the PPMI study. Indeed, two of the 24 SNPs, one in CYP7B1 (rs118111353) and the other one in CYP27A1 (rs74446825), were localized within a region of open chromatin in differentiated neurons. Interestingly, all iPSC lines with open chromatin in rs118111353 showed the reference allele. As all four P450, CYP46A1, CYP39A1, CYP27A1 and CYP7B1, are expressed in dopaminergic neurons, we discuss further functional studies to connect SNPs in regulatory regions with gene expression levels. Finally, potential possibilities for personalized therapeutic treatment of patients with SNPs in the four investigated P450 are discussed.

Frequency of rs1051338 and rs116928232 Variants in Individuals from Northwest Mexico.

LIPA, situated on chromosome 10q23.2-q23.3, encodes the enzyme lysosomal acid lipase (LAL) (EC 3.1.1.13). Genetic alterations in LIPA lead to lysosomal acid lipase deficiency (LALD), an inborn error causing lipid metabolism anomalies and impairing cholesterol and triacylglyceride degradation. Over 40 LIPA variants have been documented, yet this study focuses on just two. The rs1051338 variant (NM_000235:c.46A>C) affects the signal peptide in Exon 2, whereas rs116928232, located in Exon 8, alters the splice site (NM_000235:c.894G>A), impacting lysosomal acid lipase activity. Considering the diverse clinical manifestations of LALD and the rising hepatic steatosis prevalence in Mexican population, mainlydue to diet, these variants were investigated within this demographic to uncover potential contributing factors. This study aimed to reveal the frequency of rs1051338 and rs116928232 among healthy mestizo individuals in Northwest Mexico, marking a significant genetic exploration in this demographic. Three hundred ten healthy mestizo individuals underwent PCR-RFLP analysis for both variants, and Sanger sequencing was performed for variant rs116928232. Bioinformatic analysis was also performed to predict protein changes. Allele frequencies for rs1051338 (FA = 0.39, p value = 0.15) and rs116928232 (FA = 0.0016, p value = 0.49) aligned with reported data, while bioinformatic analysis allowed us to identify the protein alteration observed in both variants; finally, the variants showed no linkage between them (normalized D' = 1.03, p value = 0.56). Allelic frequencies closely matched reported data, and protein structure analysis confirmed variant impacts on LAL enzyme function. Notably, this study marks the first analysis of rs1051338 and rs116928232 in a healthy Mexican mestizo population.

12C6+ Irradiation Mutation for the Selection and Breeding of High-Cholesterol-Lowering Lactobacillus

A high-throughput screening method for the selection of lactic acid bacteria with high cholesterol-degrading ability was established using the 12C6+ beam irradiation mutagenesis technique. This study aimed to breed excellent strains with high cholesterol-degrading capability and explore the mechanisms of cholesterol degradation by these strains. The mutated strains JMR-01 (Lactobacillus reuteri) and SN-2 (Lactobacillus rhamnosus) after irradiation were primarily screened for high cholesterol-degrading strains by optimizing the KENJI culture medium with colony diameter as an indicator. The primary screened strains were further screened using ammonium ferric sulfate method. The mechanisms of cholesterol degradation by lactic acid bacteria were further investigated by determining the total cholesterol degradation rate, the cholesterol degradation rate through co-precipitation and absorption. The results showed that after primary and secondary screening, four mutant strains with high cholesterol-degrading ability (JMR-01-1, JMR-01-2, SN-2-1, SN-2-9) were selected. The cholesterol-degrading ability of strain JMR-01-1 was increased by 9.7% compared to the original strain JMR-01 (P<0.05), and the cholesterol-degrading ability of strain SN-2-1 was increased by 11.5% compared to the original strain SN-2. The total cholesterol degradation rate, co-precipitation, and absorption of cholesterol degradation rate of the four mutant strains showed no significant difference in the co-precipitation effect on cholesterol compared to the original strains, but the relative absorption rate was significantly increased. This study optimized the screening method for cholesterol-degrading strains and demonstrated through the exploration of different cholesterol degradation pathways by lactic acid bacteria that this screening method can select strains with high cholesterol absorption capacity.

Lemon Flavonoid Extract Eriomin Improves Pro/Antioxidant Status and Interferes with Cholesterol Metabolism without Affecting Serum Cholesterol Levels in Aged Rats.

This study aimed to assess the antioxidant capacity of lemon flavonoid extract Eriomin® (LE) and its impact on cholesterol metabolism in the context of healthy aging. We orally treated 24-month-old male Wistar rats with an LE (40 mg/kg) suspended in 0.3 mL of sunflower oil. At the same time, control groups received an equal volume of sunflower oil (CON) or remained untreated (ICON) daily for 4 weeks. We examined LE's effects on superoxide dismutase and catalase- and glutathione-related enzyme activities, the concentration of lipid peroxides and protein carbonyls, total oxidant status (TOS) and antioxidant status (TAS), and oxidative stress index (OSI) in the liver, jejunum, and ileum. We also measured total cholesterol, its biosynthetic precursors (lanosterol, lathosterol, desmosterol), its degradation products (bile acid precursors) in the serum, liver, jejunum, and ileum, and serum phytosterols (intestinal absorption markers). LE reduced TOS, TAS, and OSI (p < 0.05) compared with control values, indicating its consistent antioxidant action in all examined organs. LE lowered hepatic desmosterol (p < 0.05) while also reducing 7α- and 24-hydroxycholesterol levels in the liver and ileum (p < 0.01). Serum cholesterol, hepatic gene expression, and the immunostaining intensity of CYP7A1 were unchanged. In conclusion, LE exerted non-enzymatic antioxidant effects and reduced cholesterol degradation, reducing its biosynthesis products, thereby maintaining serum cholesterol levels.

Characterization and potential lipid-lowering effects of lactic acid bacteria isolated from cats.

This study aimed to study the characterization and the potential lipid-lowering effects of new isolated lactic acid bacteria from the feces of healthy adult cats. We collected 85 cat fecal samples, isolated, screening lactic acid bacteria strains from samples, and investigated their in vitro and in vivo biological properties. A total of 221 lactic acid bacteria strains were isolated from 85 cat fecal samples. Sixteen strains with calcium dissolution rings greater than 1 mm were identified and selected for further characterization. Three lactic acid bacteria strains, Lactobacillus plantarum L-27-2, Pediococcus lactis L-14-1, and Enterococcus faecium, were identified as showing the most promising rates of cholesterol degradation (greater than 20%) and bacteriostatic radius (over 15 mm). These three strains exhibited robust growth and adherence to epithelial cells, along with adaptability to low pH (greater than 70%) and high bile salt conditions (greater than 60%), and remarkable cholesterol degradation and anti-pathogen activity. Sixteen mice were fed a high-fat diet (HFD) from 4 to 8 weeks of age, while a control group of the same size received a normal diet (ND). At 8 weeks of age, serum, feces and adipose tissue were collected. The results showed that, compared with mice fed an HFD diet alone, all mice fed an HFD diet plus lactic acid bacteria could decrease weight gain. P < 0.05 and the pathological changes of adipose tissue were alleviated. In addition, mice fed L-14-1 and F203 showed abdominal fat accumulation decreased (P < 0.05). Mice fed L-27-2 showed serum and liver triglyceride (TG) decreased (P < 0.05) and mice fed F203 showed serum high density lipoprotein cholesterol (HDL-C) increased (P < 0.01). mice fed L-27-2 and L-14-1 showed inflammatory cytokines (IL-6) was decreased (P < 0.01) Analysis of the fecal microbiota of mice fed these three lactic acid bacteria strains revealed alterations in the gut microbial community. There were common changes in intestinal microbes in mice fed these three lactic acid bacteria: (1) Bacteroides decreased; (2) Myxococcus increased; (3) Lachnoclostridium decreased. The microbes mentioned are all part of the core intestinal flora. This study provided three potential lactic acid bacteria for alleviating animal obesity and inflammation.

Gemfibrozil-Platinum(IV) Precursors for New Enhanced-Starvation and Chemotherapy In Vitro and In Vivo.

A brand-new enhanced starvation is put forward to trigger sensitized chemotherapy: blocking tumor-relation blood vessel formation and accelerating nutrient degradation and efflux. Following this concept, two cisplatin-like gemfibrozil-derived Pt(IV) prodrugs, GP and GPG, are synthesized. GP and GPG had nanomolar IC50 against A2780 cells and higher selectivity against normal cells than cisplatin. Bioactivity results confirmed that GP and GPG highly accumulated in cells and induced DNA damage, G2-phase arrest, and p53 expression. Besides, they could increase ROS and MDA levels and reduce mitochondrial membrane potential and Bcl-2 expression to promote cell apoptosis. In vivo, GP showed superior antitumor activity in A2780 tumor-bearing mice with no observable tissue damage. Mechanistic studies suggested that highly selective chemotherapy could be due to the new enhanced starvation effect: blocking vasculature formation via inhibiting the CYP2C8/EETs pathway and VEGFR2, NF-κB, and COX-2 expression and cholesterol efflux and degradation acceleration via increasing ABCA1 and PPARα.

The role of cholesterol and its oxidation products in tuberculosis pathogenesis.

Mycobacterium tuberculosis causes tuberculosis (TB), one of the world's most deadly infections. Lipids play an important role in M. tuberculosis pathogenesis. M. tuberculosis grows intracellularly within lipid-laden macrophages and extracellularly within the cholesterol-rich caseum of necrotic granulomas and pulmonary cavities. Evolved from soil saprophytes that are able to metabolize cholesterol from organic matter in the environment, M. tuberculosis inherited an extensive and highly conserved machinery to metabolize cholesterol. M. tuberculosis uses this machinery to degrade host cholesterol; the products of cholesterol degradation are incorporated into central carbon metabolism and used to generate cell envelope lipids, which play important roles in virulence. The host also modifies cholesterol by enzymatically oxidizing it to a variety of derivatives, collectively called oxysterols, which modulate cholesterol homeostasis and the immune response. Recently, we found that M. tuberculosis converts host cholesterol to an oxidized metabolite, cholestenone, that accumulates in the lungs of individuals with TB. M. tuberculosis encodes cholesterol-modifying enzymes, including a hydroxysteroid dehydrogenase, a putative cholesterol oxidase, and numerous cytochrome P450 monooxygenases. Here, we review what is known about cholesterol and its oxidation products in the pathogenesis of TB. We consider the possibility that the biological function of cholesterol metabolism by M. tuberculosis extends beyond a nutritional role.

Exposure of Mycobacterium tuberculosis to human alveolar lining fluid shows temporal and strain-specific adaptation to the lung environment

Upon infection, Mycobacterium tuberculosis (M.tb) reaches the alveolar space and comes in close contact with the lung mucosa or human alveolar lining fluid (ALF) for an uncertain period of time prior to its encounter with alveolar cells. We showed that homeostatic ALF hydrolytic enzymes modify the M.tb cell envelope, driving M.tb-host cell interactions. Still, the contribution of ALF during M.tb infection is poorly understood. Here, we exposed 4 M.tb strains with different levels of virulence, transmissibility, and drug resistance (DR) to physiological concentrations of human ALF for 15-min and 12-h, and performed RNA sequencing. Gene expression analysis showed a temporal and strain-specific adaptation to human ALF. Differential expression (DE) of ALF-exposed vs. unexposed M.tb revealed a total of 397 DE genes associated with lipid metabolism, cell envelope and processes, intermediary metabolism and respiration, and regulatory proteins, among others. Most DE genes were detected at 12-h post-ALF exposure, with DR-M.tb strain W-7642 having the highest number of DE genes. Interestingly, genes from the KstR2 regulon, which controls the degradation of cholesterol C and D rings, were significantly upregulated in all strains post-ALF exposure. These results indicate that M.tb-ALF contact drives initial bacterial metabolic and physiologic changes, which may have implications in the early events of M.tb infection.

Beneficial effects of sour pork fermented with a Lactobacillus plantarum strain on lowering blood lipids in mice

As a type of main meat food, pork is widely accepted, and has been used over a long time by residents in China. However, its high cholesterol is the primary unfavourable factor that influences human health. The reduction of cholesterol content in pork through fermentation using probiotics may be a promising candidate approach. In the present work, the probiotic strains were obtained from traditional naturally fermented foods, and identified via monoclonal culture, morphological observation, biochemical detection, and 16S rRNA sequencing. The dominant strain was used to produce sour pork, which was applied to regulate lipid metabolism in mice. Results showed that a total of 31 strains were identified from the traditional naturally fermented foods, of which four strains had similar characteristics to Lactobacillus plantarum, and displayed good capacity for cholesterol degradation with metabolism efficiencies of 75.31% (DC2 strain), 68.01% (PC1 strain), 60.49% (SC1 strain), and 58.02% (DC1 strain). The dominant strain DC2 had 99.93% homology with L. plantarum. Following fermentation with DC2 strain (4 × 106 CFU/g inoculum size, 20-day incubation), the cholesterol in sour pork was significantly reduced with 77.2% efficiency. The serum lipids in mice that were fed with sour pork were significantly lower than those in mice that were fed with normal pork. In summary, a L. plantarum strain with good capacity for cholesterol degradation was obtained (CCTCC NO: M 2019121). The strain can be used to produce fermented sour pork. Dietary sour pork may be beneficial for lowering blood lipid levels.

Improved production of Bacillus subtilis cholesterol oxidase by optimization of process parameters using response surface methodology

BackgroundCholesterol oxidase has numerous biomedical and industrial applications. In the current study, a new bacterial strain was isolated from sewage and was selected for its high potency for cholesterol degradation (%) and production of high cholesterol oxidase activity (U/OD600). ResultsBased on the sequence of 16S rRNA gene, the bacterium was identified as Bacillus subtilis. The fermentation conditions affecting cholesterol degradation (%) and the activity of cholesterol oxidase (U/OD600) of B. subtilis were optimized through fractional factorial design (FFD) and response surface methodology (RSM). According to this sequential optimization approach, 80.152% cholesterol degradation was achieved by setting the concentrations of cholesterol, inoculum size, and magnesium sulphate at 0.05 g/l, 6%, and 0.05 g/l, respectively. Moreover, 85.461 U of cholesterol oxidase/OD600 were attained by adjusting the fermentation conditions at initial pH, 6; volume of the fermentation medium, 15 ml/flask; and concentration of cholesterol, 0.05 g/l. The optimization process improved cholesterol degradation (%) and the activity of cholesterol oxidase (U/OD600) by 139% and 154%, respectively. No cholesterol was detected in the spectroscopic analysis of the optimized fermented medium via gas chromatography-mass spectroscopy (GC–MS). ConclusionThe current study provides principal information for the development of efficient production of cholesterol oxidase by B. subtilis that could be used in various applications.

Abstract 15325: SAH-Responsive Genes Mediate Worsening Metabolic Crosstalk Between Homocysteine-Methionine, Glucose-Related &amp;Lt Lipid Metabolic Systems

Introduction: Hyperhomocysteinemia (HHcy) is an independent risk factor for cardiovascular disease, and other chronic disease. We recently proposed that the homocysteine-methionine (H-M) cycle is a metabolic sensor system responsible for pathological signaling in chronic disease. We identified metabolic worsening crosstalk between HHcy, hyperglycemia and hyperlipidemia in concomitant mouse disease models and in human coronary artery disease. Research Question: What is the molecular mechanism mediating worsening metabolic crosstalk in concomitant metabolic disorders? Methods: mRNA levels of 324 metabolic genes in three metabolic systems were examined, including 119 for H-M, 95 for glucose-related and 110 genes for lipid in 21 human and 20 mouse organs. Organs were classified based on specific metabolic gene expression. Hcy-metabolites were measured in 6 mouse organs (heart, liver, brain, lung, spleen, and kidney) by LS-ESI-MS/MS. Hcy-metabolites responsive genes were identified by linear regression analyses between metabolites and gene expression levels and used to establish Hcy-metabolites responsive metabolic networks. Results: We identified 7 Hcy-, 64 SAH- and 18 SAM:SAH-responsive genes and 3 pathological SAH-responsive cross-talking metabolites (serine, taurine and glycine 3 phosphate). SAH-responsive genes associated with activated upper glycolysis and suppressed lower glycolysis leading to glycolysis-related redox imbalance. A negative correlation between SAH and serine synthesis enzyme PHGDH suggests that increased SAH may suppress serine synthesis which leads to the impairment of transsulfuration, and taurine reduction. SAH-related taurine reduction can further negatively impact cholesterol (CH) degradation. SAH-related upper glycolysis activation leads to the increase of G3P synthesis, which is a substrate for triglyceride (TG) synthesis. In addition, SAH levels may activate fatty acid uptake and β-oxidation leading to elevated acetyl-CoA, CH and TG synthesis. Conclusion: H-M cycle is a metabolic sensor system. Production change of serine, taurine and G3P via SAH-responsive metabolic genes can be the key cross-talking metabolites responsible for worsening metabolic crosstalk in metabolic syndrome.

Transcriptomic profile of the most successful Mycobacterium tuberculosis strain in Aragon, the MtZ strain, during exponential and stationary growth phases.

The Mycobacterium tuberculosis Zaragoza (MtZ) strain has produced the largest outbreak in Aragon Community, Spain, as it started in the 90s and continues today, contributing with more than 240 cases of tuberculosis, around 10% of the TB cases. Previous studies were carried out to identify the reasons for its successful spread capacity among the population. Some single nucleotide polymorphisms (SNPs) in genes considered as virulence factors were found, as well as an oversecretion of PE_PGRS factors. Different copies of IS6110 have been localized, and a transcriptomic study was carried out to study how the different IS6110 copies could affect the adjacent gene expression. In this work, we analyzed the transcriptomic profile of the MtZ, strain trying to delve into the mechanisms behind its success. We found that ESX-1 system is upregulated during both exponential and stationary growth phases in vitro, as well as the virulence pathways of cholesterol degradation and peptidoglycan biosynthesis. In addition, MtZ has the iron uptake genes downregulated in exponential growth phase (mycobactin biosynthesis and IrtAB transporters), while the iron storage could be upregulated. The desA3 gene transcription was interrupted by the insertion of an IS6110 copy. This gene codes for a stearoyl-coenzyme A (CoA) Δ9 desaturase involved in the production of oleic acid, which is necessary for the metabolism of the bacteria. In spite of this, although MtZ strain has an initial long lag phase growing in liquid media, it is capable of achieving normal growth, suggesting the existence of an alternative route for obtaining oleic acid. IMPORTANCE Aragon Community suffered, during the first years of the beginning of this century, a large outbreak caused by the MtZ strain, producing more than 240 cases to date. MtZ strain and the outbreak have been previously studied from an epidemiological and molecular point of view. In this work, we analyzed the transcriptomic profile of the strain for better understanding of its success among our population. We have discovered that MtZ has some upregulated virulence pathways, such as the ESX-1 system, the cholesterol degradation pathway or the peptidoglycan biosynthesis. Interestingly, MtZ has downregulated the uptake of iron. Another special feature of MtZ strain is the interruption of desA3 gene, essential for producing oleic acid. Although the strain takes a long time to grow in the initial culture media, eventually it is able to reach normal optical densities, suggestive of the presence of another route for obtaining oleic acid in Mycobacterium tuberculosis.

Relay run property of immobilized cholesterol oxidase on magnetic biochar for sterol removal

This study investigated the biocatalytic performance of immobilized cholesterol oxidase (CHOD) on magnetite-based carbon (MBC) for degrading cholesterol. The results showed that MBC-CHOD exhibited higher activity and good affinity towards substrate compared to free enzyme and other immobilized enzymes. Mass spectra analysis revealed that MBC-CHOD damaged the main structure of cholesterol, benefitting the further biological treatment. The study proposes a Fenton process mechanism by which H2O2 is transferred to free radicals such as ·OH under acidic conditions, promoting further substrate degradation. This suggests that MBC-CHOD has a relay run property leading to high degradation of cholesterol. Molecular docking indicates that cholesterol preferentially binds to TYR-28 residue and LYS-138 residue in CHOD through hydrogen bonds. Overall, MBC-CHOD proved to be a promising candidate for efficient and sustainable cholesterol degradation.

SNPs in cytochrome P450 genes decide on the fate of individuals with genetic predisposition to Parkinson's disease.

Parkinson's disease (PD) is one of the most frequent neurological diseases affecting millions of people worldwide. While the majority of PD cases are of unknown origin (idiopathic), about 5%-10% are familial and linked to mutations in different known genes. However, there are also people with a genetic predisposition to PD who do not develop the disease. To elucidate factors leading to the manifestation of PD we compared the occurrence of single nucleotide polymorphisms (SNPs) in various cytochrome P450 (P450) genes in people with a genetic predisposition and suffering from PD (GPD) to that of people, who are genetically predisposed, but show no symptoms of the disease (GUN). We used the PPMI (Parkinson's Progression Markers Initiative) database and the gene sequences of all 57 P450s as well as their three redox partners. Corresponding odds ratios (OR) and confidence intervals (CI) were calculated to assess the incidence of the various SNPs in the two groups of individuals and consequently their relation to PD. We identified for the first time SNPs that are significantly (up to 10fold!) over- or under-represented in GPD patients compared to GUN. SNPs with OR > 5 were found in 10 P450s being involved in eicosanoid, vitamin A and D metabolism as well as cholesterol degradation pointing to an important role of endogenous factors for the manifestation of PD clinical symptoms. Moreover, 12 P450s belonging to all P450 substrate classes as well as POR have SNPs that are significantly under-represented (OR < 0.2) in GPD compared to GUN, indicating a protective role of those SNPs and the corresponding P450s regarding disease advancement. To the best of our knowledge our data for the first time demonstrate an association between known PD predisposition genes and SNPs in other genes, shown here for different P450 genes and for their redox partner POR, which promote the manifestation of the disease in familial PD. Our results thus shed light onto the pathogenesis of PD, especially the switch from GUN to GPD and might further help to advance novel strategies for preventing the development or progression of the disease.

Periodic Appraisal of Cholesterol Degradation by Two Bacteria Isolates from 'Iru' (Fermented Parkia Biglobosa)

This study was carried out to evaluate the incubation period on cholesterol degrading ability of bacteria isolated from 'iru' (fermented Parkia biglobosa seeds). The total bacterial count after culturing 'iru' (Parkia biglobosa) on nutrient agar were 7.30, 8.13, 7.28 and 8.11 respectively. The isolates were identified as Bacillus cereus and Bacillus subtilis. The isolates were screened on a salt agar of 1% cholesterol. Some of the isolates utilized cholesterol and grew well. IRUA and IRUC had the highest growth on the minimal salt cholesterol agar, followed by IRUB. The growth of 2IRUB was very low while IRUD, 2IRUA, 2IRUC, 2IRUD had no growth on the nutrient agar. Bacillus subtilis and Bacillus cereus degraded cholesterol maximally as 59.57mmol/L and 43.51mmol/L respectively. The cholesterol concentration degraded greatly with incubation period showing that there were high activities of the microorganisms and maximum cholesterol degradation was recorded on the 7th day. This study showed that the cholesterol degrading bacteria from Parkia biglobosa are endowed with the capability to degrade cholesterol and are good sources of cholesterol oxidase exploited for potential biomedical and industrial applications.

In vitro screening and probiotic evaluation of anti-obesity and antioxidant lactic acid bacteria

In this study, an in vitro screening method for lactic acid bacteria (LAB) with anti-obesity and antioxidant effects was developed. We first screened high cholesterol-lowering LAB from the traditional fermented food water kefir. Among the screened LAB, there were 12 strains with a cholesterol degradation rate over of 60%. The 12 strains were ranked from highest to lowest cholesterol degradation ability, and the top five strains were selected for subsequent tests to determine their ability to lower triglycerides, inhibit α-amylase and α-glucosidase, and LAB with great anti-obesity and antioxidant effects were screened in vitro by determining their ability to inhibit Enterobacter cloacae and their antioxidant activity. A comprehensive evaluation model was then combined with principal component analysis: Y = 0.513Y1+0.229Y2+0.137Y3+0.121Y4. Lactobacillus rhamnosus S51, Lactobacillus plantarum S184, and Lactobacillus fermentum S7 had better anti-obesity and antioxidant potential. The cholesterol-lowering ability of S51 was 69.90%, the triglyceride-lowering ability was 53.72%, α-amylase, and α-glucosidase inhibiting ability was 69.97% and 31.49%, respectively, and Enterobacter cloacae inhibiting ability was 3.43 cm. Its IC scavenging ability of DPPH free radical was 90.47%, which was equivalent to 27.98 μg/mL Trolox standard solution, the CFE scavenging capacity for hydroxyl radicals was 60.99%, and the total reducing power of CFE was equivalent to 513.37 μmol/L l-cysteine hydrochloride. S51, S184, and S7 were eligible for probiotic conditions. The results of the study provide a theoretical basis for the screening of LAB with anti-obesity and antioxidant effects in vitro.

Shenlian extract protected ox-LDL-loaded macrophages against ER stress by promoting LAL-LXRα mediated cholesterol flux

Ethnopharmacological relevanceShenlian (SL) extract is consisted of extracts from Salvia miltiorrhiza Bunge and Andrographis paniculata (Burm.f.) Nees, two herbs commonly used in Chinese clinical formula to treat atherosclerosis by removing blood stasis and clearing away heat. Pharmacologically, the anti-atherosclerotic effects of these two herbs are related to unresolved inflammation and the macrophage anergy or apoptosis in lesions led by the lipid flux blockage and ER stress. However, the deeper understanding of SL extract in protecting macrophage in plaques remains unknown. Aim of the studyThis study aimed to investigate the underlying mechanism of SL extract in protecting ER-stressed macrophages from apoptosis in atherosclerosis. MethodsThe ApoE−/− atherosclerotic mice model and ox-LDL loaded macrophages model were established to assess the effect of SL extract on ER stress in vivo and in vitro. Key markers related to ER stress in plaque were determined by immunohistochemical staining. Proteins involved in apoptosis and ER stress in macrophages loaded by ox-LDL were assessed by Western blot. ER morphology was observed by electron microscope. Lipid flux was temporally and quantitatively depicted by Oil red staining. The LAL and LXRα were blocked by lalistat and Gsk 2033 respectively to investigate whether SL extract protected the function of macrophages by the activation of LAL-LXRα axis. ResultsOur study reported that, in ApoE−/− atherosclerotic mice, SL extract effectively relieved ER stress of carotid artery plaque. In lipid-overloaded macrophage models, SL extract significantly alleviated ER stress by promoting cholesterol degradation and efflux, which finally prevented apoptosis of foam cells induced by ox-LDL. Blockage of ER stress by 4-Phenylbutyric acid (4-PBA), an inhibitor of Endoplasmic Reticulum (ER) stress, largely attenuated the protective effects of SL extract on macrophage. By utilizing the selective antagonists against both LAL and LXRα, this study further revealed that the beneficial effects of SL extract in macrophages was dependent on the proper functionalization of LAL-LXRα axis. ConclusionsBy highlighting the therapeutic significance of macrophage protection in resolving atherosclerosis inflammation, our study pharmacologically provided convincing mechanistic evidence of SL extract in the activation LAL-LXRα axis and revealed its promising potential in the promotion of cholesterol turnover and prevention of ER stress induced apoptosis in lipid-loaded macrophages.

Targeting FABP4 in elderly mice rejuvenates liver metabolism and ameliorates aging-associated metabolic disorders

IntroductionAging is characterized by progressive metabolic dyshomeostasis that increases morbidity and mortality. Solutions for optimizing healthy aging are challenged by lacking appropriate biomarkers. Moreover, druggable targets to rejuvenate the aging-associated metabolic phenotypes remain unavailable. MethodsProteomics analysis was performed in a cohort of young and elderly adults. Circulating levels of insulin-like growth factor 1 (IGF-1) and fatty acid binding protein 4 (FABP4) were evaluated by ELISA. FABP4 was silenced in elderly mice by adeno-associated virus. Metabolic activities were measured by metabolic cages. Cognitive function was evaluated by Morris water maze. Glucose and lipid metabolism were evaluated by biochemistry assays with blood samples. RNA-seq in mouse liver was performed for transcriptome analysis. ResultsAmong 9 aging-sensitive proteins shared by both male and female, FABP4 was identified as a reliable aging biomarker in both human and mouse. Silencing FABP4 in elderly mice significantly rejuvenated the aging-associated decline in metabolic activities. FABP4 knockdown reversed the aging-associated metabolic disorders by promoting degradation of cholesterol and fatty acids, while suppressing gluconeogenesis. Transcriptome analysis revealed a restoration of the pro-aging gene reprogramming towards inflammation and metabolic disorders in the liver after FABP4 knockdown. FABP4 overexpression promoted human LO2 cell senescence. Moreover, administration of an FABP4 inhibitor BMS309403 delivered metabolic benefits in elderly mice. ConclusionOur findings demonstrate FABP4 as a reliable aging biomarker as well as a practicable target to improve healthy aging in the elderly.

The function of steroid terminal monooxygenase in cholesterol or phytosterol degradation by the Mycobacterium neoaurum NRRL B-3805

Cholesterol and phytosterols can be served as the substrates for Actinobacteria to produce the precursors used in the synthesis of steroidal medicines. Cytochrome P450s from CYP125 and CYP142 families initiates the metabolism of cholesterol and/or phytosterols. In this study, we demonstrate the functional redundancy and substrate preference of five cytochrome CYP125s and CYP142s by the unmarked cyp genes deletion strains of Mycobacterium neoaurum NRRL B-3805. CYP125-3 was found to be responsible for the catalytic hydroxylation of both cholesterol and the C24-branched side-chains of phytosterols, while CYP142-2 can only oxidize cholesterol and/or cholest-4-en-3-one. The strain harboring the CYP125-3 produced 6.47 g l− 1 and 5.13 g l− 1 of 4-androstene-3,17-dione (AD) when 15 g l− 1 cholesterol or 17.5 g l− 1 phytosterols used, respectively. While, the strain harboring the CYP142-2 produced 7.10 g l− 1 AD when 15 g l− 1 cholesterol used. The other three cytochrome P450s have a smaller contribution for the degradation of cholesterol and phytosterols. CYP125-3 has the hydroxylation activity on more substrates than that of CYP142-2. This study provided possible guidance to improve the efficiency of AD and other metabolites production from different kinds of sterols by the genetic manipulation.

A CYPome-wide study reveals new potential players in the pathogenesis of Parkinson's disease.

Genetic and environmental factors lead to the manifestation of Parkinson's disease (PD) but related mechanisms are only rudimentarily understood. Cytochromes P450 (P450s) are involved in the biotransformation of toxic compounds and in many physiological processes and thus predestinated to be involved in PD. However, so far only SNPs (single nucleotide polymorphisms) in CYP2D6 and CYP2E1 have been associated with the susceptibility of PD. Our aim was to evaluate the role of all 57 human P450s and their redox partners for the etiology and pathophysiology of PD and to identify novel potential players which may lead to the identification of new biomarkers and to a causative treatment of PD. The PPMI (Parkinson's Progression Markers Initiative) database was used to extract the gene sequences of all 57 P450s and their three redox partners to analyze the association of SNPs with the occurrence of PD. Applying statistical analyses of the data, corresponding odds ratios (OR) and confidence intervals (CI) were calculated. We identified SNPs significantly over-represented in patients with a genetic predisposition for PD (GPD patients) or in idiopathic PD (IPD patients) compared to HC (healthy controls). Xenobiotic-metabolizing P450s show a significant accumulation of SNPs in PD patients compared with HC supporting the role of toxic compounds in the pathogenesis of PD. Moreover, SNPs with high OR values (>5) in P450s catalyzing the degradation of cholesterol (CYP46A1, CY7B1, CYP39A1) indicate a prominent role of cholesterol metabolism in the brain for PD risk. Finally, P450s participating in the metabolism of eicosanoids show a strong over-representation of SNPs in PD patients underlining the effect of inflammation on the pathogenesis of PD. Also, the redox partners of P450 show SNPs with OR > 5 in PD patients. Taken together, we demonstrate that SNPs in 26 out of 57 P450s are at least 5-fold over-represented in PD patients suggesting these P450s as new potential players in the pathogenesis of PD. For the first time exceptionally high OR values (up to 12.9) were found. This will lead to deeper insight into the origin and development of PD and may be applied to develop novel strategies for a causative treatment of this disease.