Chemical profiling of Sanjin tablets and exploration of their effective substances and mechanism in the treatment of urinary tract infections.

Introduction: Sanjin tablets (SJT) are a well-known Chinese patent drug that have been used to treat urinary tract infections (UTIs) for the last 40 years. The drug consists of five herbs, but only 32 compounds have been identified, which hinders the clarification of its effective substances and mechanism. Methods: The chemical constituents of SJT and their effective substances and functional mechanism involved in the treatment of UTIs were investigated by using high performance liquid chromatography-electrospray ionization-ion trap-time of flight-mass spectrometry (HPLC-ESI-IT-TOF-MSn), network pharmacology, and molecular docking. Results: A total of 196 compounds of SJT (SJT-MS) were identified, and 44 of them were unequivocally identified by comparison with the reference compounds. Among 196 compounds, 13 were potential new compounds and 183 were known compounds. Among the 183 known compounds, 169 were newly discovered constituents of SJT, and 93 compounds were not reported in the five constituent herbs. Through the network pharmacology method, 119 targets related to UTIs of 183 known compounds were predicted, and 20 core targets were screened out. Based on the "compound-target" relationship analysis, 94 compounds were found to act on the 20 core targets and were therefore regarded as potential effective compounds. According to the literature, 27 of the 183 known compounds were found to possess antimicrobial and anti-inflammatory activities and were verified as effective substances, of which 20 were first discovered in SJT. Twelve of the 27 effective substances overlapped with the 94 potential effective compounds and were determined as key effective substances of SJT. The molecular docking results showed that the 12 key effective substances and 10 selected targets of the core targets have good affinity for each other. Discussion: These results provide a solid foundation for understanding the effective substances and mechanism of SJT.

Acceleration effect of galacturonic acid on acrylamide generation: evidence in model reaction systems.

BACKGROUND: Acrylamide (AA) is a potential carcinogen formed in food rich in carbohydrate during heating. Recently, AA has been found in several fruit products, such as prune juice, sugarcane molasses and canned black olives. This study focused on the role of galacturonic acid (GalA), the main acid hydrolysis product of fruit pectin, in AA formation in three model systems - asparagine (Asn)/glucose (Glc), Asn/GalA, and Asn/Glc/GalA - during heating under different pH values (pH 3.8-7.8), Glc concentration (0-0.1 mol L-1 ), molar ratio of substrates (Asn/Glc = 1:1, 0.025-0.5 mol L-1 ) and temperature (120-180 °C) for 30 min, respectively. RESULTS: The results suggested that the addition of 0.1 mol L-1 GalA strongly accelerated AA formation in a manner dependent on pH value and temperature (P < 0.05). AA concentration under different Glc concentration and molar ratio of substrates suggested that GalA was more reactive than Glc when reacted with Asn. Furthermore, the Amadori rearrangement product/Schiff base/oxazolidine-5-one were identified as the intermediates formed in the Asn/GalA model system using ultra-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry. CONCLUSION: The results suggested that Maillard reaction between Asn and GalA might contribute to AA formation. This study is significant in elucidating the contribution of interaction between components for AA formation in fruit products. © 2022 Society of Chemical Industry.

Paeoniae Radix Rubra can enhance fatty acid ß-oxidation and alleviate gut microbiota disorder in α-naphthyl isothiocyanate induced cholestatic model rats.

Cholestasis is the most destructive pathological manifestation of liver disease and available treatments are very limited. Paeoniae Radix Rubra (PRR) is an important traditional Chinese drug used to treat cholestasis. This study combined targeted metabonomics, PCR array analysis, and 16S rRNA sequencing analysis to further clarify the mechanisms of PRR in the treatment of cholestasis. PRR conspicuously reversed the elevation of fatty acids (FFA 14:0 and other 14 fatty acids) and the decrease of organic acids (pyruvic acid and citric acid) in a cholestatic model induced by α-naphthyl isothiocyanate (ANIT). Eight elevated amino acids (L-proline, etc.) and five elevated secondary bile acids (taurohyodeoxycholic acid, etc.) in model rats were also reduced by PRR. Pathway analysis revealed that PRR significantly alleviated eight pathways (ß-alanine metabolism). Furthermore, we found that PRR significantly reversed the decrease of Cpt1a, Hadha, Ppara, and Slc25a20 (four genes relevant to fatty acid ß-oxidation) mRNAs caused by ANIT, and PRR conspicuously decreased nine acylcarnitines (the forms of fatty acids into mitochondria for ß-oxidation) that increased in model rats. These results indicate that PRR could enhance fatty acid ß-oxidation, which may be the way for PRR to reduce the levels of 15 fatty acids in the serum of model rats. 16S rRNA sequencing analysis revealed that PRR alleviated gut microbiota disorders in model rats, including upregulating four genera (Coprococcus, Lactobacillus, etc.) and downregulating four genera (Bacteroides, Escherichia, etc.). As the relative abundance of these eight genera was significantly correlated with the levels of the five secondary bile acids (deoxycholic acid, taurolithocholic acid, etc.) reduced by PRR, and Bacteroides and Escherichia were reported to promote the production of secondary bile acid, we inferred that the downregulation of PRR on five secondary bile acids in model rats was inseparable from gut microbiota. Thus, the gut microbiota also might be a potential pharmacological target for the anticholestatic activity of PRR. In conclusion, we consider that the mechanisms of PRR in treating cholestasis include enhancing fatty acid ß-oxidation and alleviating gut microbiota disorders.

Metabolism of Paeoniae Radix Rubra and its 14 constituents in mice.

Objective: Paeoniae Radix Rubra (PRR) is a commonly used traditional Chinese medicine with the effects of clearing away heat, cooling the blood, and relieving blood stasis. To 1) elucidate the metabolites and metabolic pathways of PRR and its 14 main constituents in mice and 2) reveal the possible origins of the known effective forms of PRR and their isomers, the metabolism of PRR in mice was systematically studied for the first time. Methods: PRR and its 14 constituents were administered to mice by gavage once a day for seven consecutive days, respectively. All urine and feces were collected during the 7 days of dosing, and blood was collected at 1 h after the last dose. Metabolites were detected and identified using high performance liquid chromatography with diode array detector and combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry (HPLC-DAD-ESI-IT-TOF-MSn). Results: In total, 23, 16, 24, 17, 18, 30, 27, 17, 22, 17, 33, 3, 8, 24, and 31 metabolites of paeoniflorin, albiflorin, oxypaeoniflorin, benzoylpaeoniflorin, hydroxybenzoylpaeoniflorin, benzoyloxypaeoniflorin, galloylpaeoniflorin, lactiflorin, epicatechin gallate, catechin gallate, catechin, ellagic acid, 3,3'-di-O-methylellagic acid, methylgallate, and PRR were respectively identified in mice; after eliminating identical metabolites, a total of 195 metabolites remained, including 8, 11, 25, 17, 18, 30, 27, 17, 21, 17, 1, 2, 8, 20, and 20 newly identified metabolites, respectively. The metabolic reactions of PRR and its 14 main constituents in mice were primarily methylation, hydrogenation, hydrolysis, hydroxylation, glucuronidation, and sulfation. Conclusion: We elucidated the metabolites and metabolic pathways of PRR and its 14 constituents (e.g., paeoniflorin, catechin, ellagic acid, and gallic acid) in mice and revealed the possible origins of the 10 known effective forms of PRR and their isomers. The findings are of great significance to studying the mechanism of action and quality control of PRR.

Mitigation effects of high methoxyl pectin on acrylamide formation in the Maillard model system.

Acrylamide (AA) is a potential carcinogen formed during the process of food heating. Pectin is natural food additive widely presented in fruits and vegetables. This study aimed at investigating the influence of the addition of high methoxyl apple pectin (esterification degree: 82.6%) on AA inhibition in the asparagine (Asn)/glucose (Glc) model system. Results showed that temperature (120-180 °C), pH value (6.0-7.2), pectin addition (0.2-1.0%, w/v), substrate concentration (0.01-0.5 M) and molar ratio of Asn/Glc (5:1-1:10) had significant influence on inhibition of pectin on AA formation. With adding 1.0% (w/v) pectin, the pH value, Glc consumption and Schiff base abundance declined in Asn/Glc model system. Moreover, heating treatment decreased the pH value, molecular weight, esterification degree and galacturonic acid content of pectin. Finally, the pectin degradation product was identified, which might compete with Glc for Asn in Maillard reaction, led to AA reduction. This study provided distinct evidence for controlling AA formation.

Simultaneous quantitation of acrylamide, 5-hydroxymethylfurfural, and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine using UPLC-MS/MS.

A robust and sensitive isotope dilution UPLC-MS/MS method was established for the simultaneous analysis of acrylamide (AA), 5-hydroxymethylfurfural (HMF), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP). A fairly good separation of three analytes was achieved within 4.5 min. High correlation coefficients (R2 > 0.9998) of the three compounds were obtained in their respective linear ranges. This method demonstrated low limits of detection (1.57 µg/L for AA, 0.61 µg/L for HMF, and 0.02 µg/L for PhIP) and limits of quantification (5.22 µg/L for AA, 2.03 µg/L for HMF, and 0.05 µg/L for PhIP). This method also demonstrated excellent quantification accuracy (99.02%-101.12%), precision (RSD < 6%), and recovery (82.83-119.92%) in the Maillard model systems and deep-fried meatballs. This work develops a fundamental method for the rapid simultaneous determination of AA, HMF, and PhIP in thermally processed foods that are both carbohydrates-rich and protein-rich, meanwhile providing technical support for the generation mechanism of various hazards.

The simultaneous inhibition of histidine on 5-hydroxymethylfurfural and acrylamide in model systems and cookies.

5-Hydroxymethylfurfural (HMF) and acrylamide (AA) are neoformed food contaminants. In this study, the simultaneous inhibition of HMF and AA by histidine (His) were investigated. In the asparagine (Asn)/glucose (Glc) model system, the inhibition ratios of HMF and AA were in the range of 28-58% and 0-71% when 20 mmol/L His was added. In cookies, His also exhibited excellent inhibition effects on both HMF and AA. At the His concentration of 2% (w/w), the inhibition ratios of HMF and AA reached 90% and 65%. Additionally, the sensory quality of cookies was not affected significantly. Qualitative results suggested that His inhibited the formation of AA by the competitive reaction between His and Asn for Glc, as well as directly eliminated the formed HMF and AA via the carbonyl-amine reaction and the Michael addition, respectively. This study revealed that His could be applied for the inhibition of HMF and AA in heated food.

Inhibition of acrylamide by glutathione in asparagine/glucose model systems and cookies.

The inhibition effects of glutathione (GSH) on acrylamide (AA) in the asparagine/glucose model systems and cookies were investigated. The formation of AA was significantly inhibited by GSH addition in the model systems. The decreasing levels of AA were in the range of 38-86%, 57-82%, and 3-41% at 120, 140, and 160 °C, respectively. In addition, the presence of GSH in cookies also inhibited the AA formation, but with no corresponding relationship between the GSH level and the inhibition ratio. The addition of 0.05 g/kg GSH in cookie dough resulted in the decreasing of AA by 48%, some improvement in cookie lightness, but no influence on cookie texture. Kinetic analysis showed that the application of GSH not only inhibited the formation of AA but also improved the elimination of AA. In the summary, GSH could be a potential inhibitor to reduce the AA formation in food during the heating process.

Role of glutathione on acrylamide inhibition: Transformation products and mechanism.

Glutathione (GSH) is a potential inhibitor for acrylamide (AA) in heated food. In the present study, the inhibition pathways of GSH on AA were investigated in the asparagine(Asn)/glucose(Glc)/GSH model system. In comparison to the Asn/Glc model system, three unique molecular ions (m/z 470, 379, and 193) were identified in the Asn/Glc/GSH model system. Those molecular ions were confirmed as the Amadori rearrangement products which formed in the reaction between Glc and GSH, as well as the addition reaction products between AA and the sulfhydryl group (-SH) of GSH and cysteine (Cys). The competition between Asn and GSH for Glc in the competitive reactions was assumed to be the major pathway. Additionally, the elimination reaction between AA and GSH or between AA and Cys also played a minor role in the inhibition of AA. The variances of precursors, intermediates, and final products provided quantitative evidence for the above pathways.

The modulation of Luffa cylindrica (L.) Roem supplementation on gene expression and amino acid profiles in liver for alleviating hepatic steatosis via gut microbiota in high-fat diet-fed mice: insight from hepatic transcriptome analysis.

Luffa cylindrica is a nutrient-dense vegetable with medical properties and can alleviate metabolic diseases. Numerous evidences demonstrated gut microbiota impacted the progress of nonalcoholic fatty liver disease (NAFLD). This study was to investigate the underlying mechanism of L. cylindrica supplementation against NALFD via gut microbiota from hepatic transcriptional and metabolic analysis. In diet-induced obese mice, we observed L. cylindrica supplementation (2 g/kg body weight) effectively alleviated high-fat diet-induced obese symptoms such as body weight, fat deposition, and insulin resistance. Notably, L. cylindrica supplementation significantly relieved hepatic steatosis and inflammation infiltration to decrease hepatic toxicity. RNA-sequencing analysis showed that 130 hepatic genes in total significantly altered responding to L. cylindrica supplementation. And signaling pathway analysis revealed that L. cylindrica supplementation down-regulated the transcriptional expressions of CD36 and Rxrg to inhibit hepatic lipid synthesis. Moreover, L. cylindrica supplementation increased the transcriptional expressions of Ass1, Cps1, Cth, Got1, Tat, and Gls2 to enhance amino acid levels (Gly, Ala, Pro, Val, Ile, Asn, Met, and Phe) and improve hepatic abnormal gluconeogenesis. Furthermore, in antibiotic-treated obese mice, L. cylindrica supplementation did not change these gene expressions along with the hepatic levels of lipid and amino acids. Taken together, L. cylindrica supplementation could effectively suppress hepatic steatosis in diet-induced obese mice through inhibiting lipid synthesis and enhancing amino acid levels in liver, which depended on gut microbiota. Thus, L. cylindrica might be one promising dietary supplementation targeting at gut microbiota to reduce NAFLD risk.

Targeting the gut microbiota by dietary nutrients: A new avenue for human health.

The gut microbiota is a complex ecosystem consisted of trillions of microbes that have co-evolved with their host for hundreds of millions of years. During the last decade, a growing body of knowledge has suggested that there is a compelling set of connections among diet, gut microbiota and human health. Various physiological functions of the host, ranging from metabolic and immune regulation to nerve and endocrine development, are possibly mediated by the structural components of microbial cell or the products of microbial metabolism, which are greatly influenced by dietary macronutrients and micronutrients. Thus, governing the production and activity of these microbial-associated small molecules and metabolites through dietary intervention may provide promising strategies for the improvement of human health and disease. In this review article, we first provide an overview of current findings about the intimate interrelationships between diet and gut microbiota. We also introduce the physiological effects of some microbial-associated small molecules and metabolites on the host as well as the detailed signaling mechanisms.

Changes of metabolites of acrylamide and glycidamide in acrylamide-exposed rats pretreated with blueberry anthocyanins extract.

Blueberry anthocyanins extract (BAE) is widely used natural antioxidant. The inhibitory effect of BAE (175 mg/kg bw/day) pretreatment on glycidamide (GA) metabolism following acrylamide (AA) (35 mg/kg bw/day) gavage for consecutive 7 days and 14 days in SD rat model were investigated in this work. AA, GA and its metabolites in urine, blood and 3 tissues were quantified using UPLC-MS/MS, respectively. The results showed pretreatment with BAE could significantly block the epoxidation of AA to GA. Compared with the control, BAE pretreatment significantly decreased the amount of GA and its mercapturic acid metabolite N-acetyl-S-(3-amino-2-hydroxy-3-oxopropyl)-cysteine (GAMA3) in rats urine (p < 0.05), and sharply decreased the accumulation of GA DNA adduct N7-(2-carbamoyl-2-hydroxyethyl) guanine (N7-GA-Gua) in liver, kidney and lung (p < 0.05). Meanwhile, significant reduction (p < 0.05) of N-(2-carbamoyl-2-hydroxyethyl) valine (GA-VAL) content was found in erythrocyte of the rats treated with BAE. This study provides a novel evidence for elucidating the chemopreventive effect of BAE on AA toxicity.

Gut microbiota promotes production of aromatic metabolites through degradation of barley leaf fiber.

Barley leaf (BL) contains abundant plant fibers, which are important substrates for the metabolism and degradation by the gut microbiota. Here we show that mice fed a diet supplemented with BL exhibited altered gut bacterial composition characterized by the enrichment of fiber-degrading bacteria Lachnospiraceae and Prevotella. Gut microbiota-mediated BL degradation promoted butyrate and propionate production. Metabolomic analysis showed increased aromatic metabolites such as ferulic acid, 3-phenylpropanoic acid, 3-hydroxyphenylacetic acid and 3-hydroxyphenylpropionic acid in feces of mice fed with BL. Finally, antibiotic treatment and anaerobic fermentation confirmed the obligate role of gut microbiota in the production of aromatic metabolites during BL degradation. Together, these findings provide insights into a gut microbiota-mediated degradation process of BL fiber components, which results in the production of microbial-associated metabolites that may exert potential active effects on host physiology.

Protection against neo-formed contaminants (NFCs)-induced toxicity by phytochemicals.

Neo-formed compounds (NFCs) are commonly found in all kinds of foods due to the complex reaction between components during processing. Acrylamide, benzo(a)pyrene and heterocyclic aromatic amines are the main types of NFCs in foods enriched with carbohydrate, fats and proteins, respectively. They have exhibited diverse toxicity, such as neurotoxicity, genotoxicity, potentially carcinogenic and reproductive toxicity. In recent years, various phytochemicals have been found to be effective in alleviation of their related toxicities both in vitro and in vivo. This review provides evidences on the protection roles of phytochemicals against the diverse toxicity induced by three NFCs. Moreover, the prevention mechanisms of phytochemicals are summarized. Three potential aspects involving excellent antioxidant activity, DNA protection and enzyme induction contribute to the successful protection mechanism. Meanwhile, the limitations from existing knowledge have been illustrated and the possible perspectives for the further study have also been considered. The information from this review would be useful to provide an easier and better way to improve human health when considering the possibility of using foods enriched with phytochemicals for prevention of the toxicity of exogenous pollutants.

Health benefits of anthocyanins and molecular mechanisms: Update from recent decade.

Anthocyanins are one of the most widespread families of natural pigments in the plant kingdom. Their health beneficial effects have been documented in many in vivo and in vitro studies. This review summarizes the most recent literature regarding the health benefits of anthocyanins and their molecular mechanisms. It appears that several signaling pathways, including mitogen-activated protein kinase, nuclear factor κB, AMP-activated protein kinase, and Wnt/ß-catenin, as well as some crucial cellular processes, such as cell cycle, apoptosis, autophagy, and biochemical metabolism, are involved in these beneficial effects and may provide potential therapeutic targets and strategies for the improvement of a wide range of diseases in future. In addition, specific anthocyanin metabolites contributing to the observed in vivo biological activities, structure-activity relationships as well as additive and synergistic efficacy of anthocyanins are also discussed.

The gut microbiota: A treasure for human health.

The interplay between the host and host-associated gut microbiota is an area of increasing interest during the recent decade. From young infants to elderly people, from primitive tribes to modern societies, accumulating evidence has suggested the association of critical physiological roles of gut microbiota in the pathogenesis of a variety of human metabolic, immunological and neurological diseases. Importantly, it appears that the relationship between the gut microbiota and disease is bidirectional, instead of causal or consequential. Personalized nutritional and therapeutic strategies targeting the gut microbiota such as prebiotics, probiotics, drugs and fecal microbiota transplantation may create a new era in the human health.

Simultaneous determination of free amino acids in Pu-erh tea and their changes during fermentation.

Pu-erh ripened tea is produced through a unique microbial fermentation process from the sun-dried leaves of large-leaf tea species (Camellia sinensis (Linn.) var. assamica (Masters) Kitamura) in Yunnan province of China. In this study, the changes of amino acid profiles during fermentation of Pu-erh tea were investigated, based on the improved HPLC-UV method with PITC pre-column derivatization for the simultaneous determination of twenty free amino acids. Results showed that aspartic acid, glutamic acid, arginine, alanine, theanine and tyrosine were the major amino acids in tea samples. Fermentation significantly influenced on the amino acid profiles. The total free amino acid contents significantly decreased during fermentation (p<0.05). Meanwhile, low amount of acrylamide were detected. Its concentration increased after 7-days' fermentation and then decreased gradually. The results provided the useful information for the manipulation of fermentation process according to the changes of amino acids and acrylamide contents in Pu-erh ripened tea.

Metabolism of Acrylamide: Interindividual and Interspecies Differences as Well as the Application as Biomarkers.

BACKGROUND: Acrylamide (AA), a known neurotoxin, has been considered to be a probable human carcinogen. The discovery of AA in many common foods in 2002 has caused worldwide attention and led to numerous research efforts on the metabolism of AA. METHODS: By collecting research literatures related to metabolism of AA, the present review not only summarized the major metabolic pathways and enzymes of AA, but also compared the interindividual and the interspecies differences of AA metabolism among humans, rats and mice. Moreover, the application of the metabolites as biomarkers for the AA exposure assessment in human population was also discussed. RESULTS: The interindividual differences of AA metabolism may be attributed to the activity and/or genetic polymorphisms of metabolic enzymes in individuals with different gender, age, smoking and alcohol status. Moreover, the metabolism of AA in humans may be more prone to the phase II conjugation with glutathione (GSH) than the phase I conversion of AA to glycidamide (GA) when compared with rats and mice. Both the hemoglobin (Hb) adducts and urinary mercapturic acid (MA) metabolites have been successfully used as biomarkers for the risk assessment of AA and new metabolic biomarkers are being developed. CONCLUSION: The genotoxic risk from AA may be determined by the balance between the phase I P450 2E1 (CYP2E1)-dependent toxification pathway to form genotoxic GA and the phase II GSH-conjugated detoxification pathway to form MA metabolite. Understanding the metabolism of AA in the body is helpful for developing effective intervention strategies to mitigate its toxicity.

The kinetics of the inhibition of acrylamide by glycine in potato model systems.

BACKGROUND: Acrylamide (AA) is a potential carcinogen which widely exists in heat-processed foods. The addition of glycine (Gly) has been shown to reduce the formation of AA. The objective of this work was to investigate the kinetics of the inhibition of AA by Gly in both asparagine (Asn)/glucose (Glc) and Asn/Glc/Gly potato model systems during heating at 160 °C, 180 °C, and 200 °C. RESULTS: The simplified two consecutive first-order kinetic model fitted well to the changes of AA in both systems. No significant difference in rate constant (kF) and apparent activation energy (EaF) was observed for AA formation between the two systems (P > 0.05). Whereas EaE and only kE at 200 °C for AA elimination in the Asn/Glc/Gly system was significantly higher than Asn/Glc system (P < 0.05). The elimination reaction between Gly and AA was confirmed by the identification of their major reaction product 2-((3-amino-3-oxopropyl)amino)acetic acid in the Asn/Glc/(15) N-Gly system. CONCLUSION: The reduction of AA by Gly is predominantly attributed to the elimination reaction between Gly and AA.

The chemoprotection of a blueberry anthocyanin extract against the acrylamide-induced oxidative stress in mitochondria: unequivocal evidence in mice liver.

Acrylamide (AA) is one of the most important contaminants occurring in heated food products. Accumulating evidence indicates that AA-induced toxicity is associated with oxidative stress and long-term exposure to AA induced mitochondria collapse and finally leads to apoptosis. Whereas anthocyanins are natural antioxidants and have a strong ability to reduce oxidative damage in vivo. This study investigates the protection of a blueberry anthocyanin extract (BAE) against AA-induced mitochondrial oxidative stress in mice models. The activities of electron transport chain complexes, oxidative status, and the structure and function of mitochondria were measured. Results showed that pretreatment with BAE markedly inhibited reactive oxygen species (ROS) formation, and prevented the successive events associated with the mitochondrial damage and dysfunction, including recovered activities of electron transport chain, ATPase and superoxide dismutase, ameliorated depolarization of mitochondrial membrane potential and membrane lipid peroxidation, reduced release of cytochrome c and protection of mitochondria against swelling. In a word, mitochondria are a key target at the organelle level for the protective effect of BAE against AA toxicity. These results will be helpful to provide new clues for a better understanding of the AA toxicity intervention mechanism and for exploring effective dietary constituents for intervention of AA toxicity.