Celastrol as an intestinal FXR inhibitor triggers tripolide-induced intestinal bleeding: Underlying mechanism of gastrointestinal injury induced by Tripterygium wilfordii.

BACKGROUND: Tripterygium wilfordii has been widely used for the treatment of rheumatoid arthritis, which is frequently accompanied by severe gastrointestinal damage. The molecular mechanism underlying the gastrointestinal injury of Tripterygium wilfordii are yet to be elucidated. METHODS: Transmission electron microscopy, and pathological and biochemical analyses were applied to assess intestinal bleeding. Metabolic changes in the serum and intestine were determined by metabolomics. In vivo (time-dependent effect and dose-response) and in vitro (double luciferase reporter gene system, DRATs, molecular docking, HepG2 cells and small intestinal organoids) studies were used to identify the inhibitory role of celastrol on intestinal farnesoid X receptor (FXR) signaling. Fxr-knockout mice and FXR inhibitors and agonists were used to evaluate the role of FXR in the intestinal bleeding induced by Tripterygium wilfordii. RESULTS: Co-treatment with triptolide + celastrol (from Tripterygium wilfordii) induced intestinal bleeding in mice. Metabolomic analysis indicated that celastrol suppressed intestinal FXR signaling, and further molecular studies revealed that celastrol was a novel intestinal FXR antagonist. In Fxr-knockout mice or the wild-type mice pre-treated with pharmacological inhibitors of FXR, triptolide alone could activate the duodenal JNK pathway and induce intestinal bleeding, which recapitulated the pathogenic features obtained by co-treatment with triptolide and celastrol. Lastly, intestinal bleeding induced by co-treatment with triptolide and celastrol could be effectively attenuated by the FXR or gut-restricted FXR agonist through downregulation of the duodenal JNK pathway. CONCLUSIONS: The synergistic effect between triptolide and celastrol contributed to the gastrointestinal injury induced by Tripterygium wilfordii via dysregulation of the FXR-JNK axis, suggesting that celastrol should be included in the quality standards system for evaluation of Tripterygium wilfordii preparations. Determining the mechanism of the FXR-JNK axis in intestinal bleeding could aid in the identification of additional therapeutic targets for the treatment of gastrointestinal hemorrhage diseases. This study also provides a new standard for the quality assessment of Tripterygium wilfordii used in the treatment of gastrointestinal disorders.

Adolescent Hypertension Induced by Obesity and the Efficacy of Comprehensive Intervention.

Context: With the development of the Chinese economy, people's quality of life has improved, obesity caused by excessive nutrition has increased among teenagers, and the age of patients with obesity-induced hypertension has become younger and younger. Objective: The study intended to analyze the factors affecting hypertension in adolescents to find methods to effectively prevent and treat hypertension in that age group. Design: The research team designed a prospective controlled study. Setting: The study took place at the First People's Hospital of Nantong in Nantong, Jiangsu, China from 2020 to 2022 years. Participants: Participants were 1000 students in Grades 7 to 9 at the Si'an middle school in Nantong, China. Intervention: From the 1000 participants, among of them 500 cases of hypertension. The research team (n = 500) participants who were obese and hypertensive and assigned them to the hypertension group, the control group (n = 500) participants with normal weights and no hypertension assigned them to a control group. Participants with obesity-induced hypertension received a comprehensive intervention. Outcome Measures: The research team used a questionnaire and a physical examination to collect data about participants' ages, heights, weights, demographic characteristics, waist circumferences, hip circumferences, and knowledge of hypertension and blood pressure and analyzed the relationships between those factors. The team measured blood pressure, blood lipids, blood glucose, and body mass index at baseline and postintervention. Results: Significant differences existed between the hypertension and control groups at general data: (1) weight-63.49 ± 13.22 kg and 52.59 ± 10.21 kg, respectively (P = .000); (2) waist circumference-75.44 ± 10.92 cm and 68.73 ± 8.15 cm, respectively (P = .001); (3) hip circumference-92.10 ± 7.98 cm and 85.95 ± 7.91 cm, respectively (P = .000); (4) body mass index (BMI)-22.12 ± 4.02 kg/m2 and 19.58 ± 3.34 kg/m2, respectively (P = .002); (5) waist-hip ratio-0.83 ± 0.08 and 0.81 ± 0.07, respectively (P = .003); and (6) waist-to-height ratio-0.46 ± 0.07 and 0.44 ± 0.06, respectively (P = .000). Only age (p=0.006), hip circumference(p=0.000), and BMI (P = .000) were significantly and positively correlated with hypertension. The regression coefficients for age, hip circumference, and BMI were 0.182, 0.062 and 0.096, respectively. The changes in the hypertension group's mean systolic blood pressure (SBP), diastolic blood pressure (DBP), BMI, body fat, trunk fat, abdominal fat, upper-limb fat, and lower-limb fat between baseline and postintervention were statistically significant. Significant decreases in the hypertension group's triglycerides, total cholesterol, and glycated albumin had occurred between baseline and postintervention (all P < .01), and that group's glucagon (P = .011) had significantly increased. No significant changes had occurred in that group's blood glucose, glycated hemoglobin, insulin, and insulin resistance index between baseline and postintervention (P < .05). Conclusions: Obesity increases the risk of hypertension, and comprehensive interventions can effectively prevent and treat adolescent hypertension.

Structure characterization of a highly branched galactan from the slug Vaginulus alte and its utilization by human gut microbiota.

The slug Vaginulus alte is used as folk medicine in China, but the structure and activities of its galactan components remain to be clarified. Here, the galactan from V. alte (VAG) was purified. The Mw of VAG was determined as ~28.8 kDa. Chemical composition analysis showed that VAG was composed of d-galactose (75 %) and l-galactose (25 %). To elucidate its precise structure, a series of disaccharides and trisaccharides were purified from mild acid hydrolyzed VAG and their structures were characterized by 1D/2D NMR spectroscopy. Based on methylation analysis and structural analysis of oligosaccharides, VAG was elucidated as a highly branched polysaccharide and mainly composed of (1 â†’ 6)- or (1 â†’ 3)-linked ß-d-galactose, and distinct (1 â†’ 2)-linked α-l-galactose. The investigation of probiotic effects in vitro revealed that VAG could promote the growth of B. thetaiotaomicron and B. ovatus, while had no effect on the growth of L. acidophilus, L. rhamnosus, B. longum subsp. infantis and B. animalis subsp. lactis, but dVAG-3 with Mw ~1.0 kDa could promote the growth of L. acidophilus. These results will provide insights into specific structures and functions of polysaccharides from the V. alte.

Crowberry inhibits cell proliferation and migration through a molecular mechanism that includes inhibition of DEK and Akt signaling in cholangiocarcinoma.

BACKGROUND: Cholangiocarcinoma (CCA) is a rare biliary adenocarcinoma related to poor clinical prognosis. Crowberry is an herbal medicine used to control inflammatory diseases and reestablish antioxidant enzyme activity. Although crowberry shows significant therapeutic efficacy in various tumors and diseases, its anticancer effects and specific molecular mechanisms in CCA are poorly understood. AIM OF THE STUDY: This study was conducted to characterize crowberry effects on CCA cells behavior. MATERIALS AND METHODS: The chemical profiles of crowberry extract was qualitatively analyzed by high-performance liquid chromatography (HPLC) and HPLC-tandem mass spectrometry. MTT, colony formation and EdU assays were performed to measure cell proliferation. The effect of crowberry treatment on CCA cell migration was assessed by wound healing and migration assays. Moreover, Hoechst staining assay and flow cytometry were performed to assess the cell apoptosis rate. Western blotting was used to assess the protein expression levels of key factors associated with apoptosis, the Akt signaling pathway, and the epithelial-mesenchymal transition. A xenograft model was established and immunohistochemical and H&E staining was performed to assess crowberry antitumor effects in vivo. RESULTS: Crowberry clearly inhibited CCA cells proliferation and migration in a dose-dependent manner and induced apoptosis in vitro. Crowberry inactivated the PI3K/Akt signaling pathway by regulating DEK in vitro and significantly inhibited tumor growth by downregulating the DEK expression in xenograft models. CONCLUSION: Crowberry inhibits CCA cells proliferation and migration through a molecular mechanism that includes inhibition of DEK and Akt signaling pathway inhibition in vitro and in vivo.

Gas purge micro solvent extraction: A rapid and powerful tool for essential oil chromatographic fingerprints.

In this work, a new an innovative micro liquid extraction technique named "gas purge micro solvent extraction" (GP-MSE) have been tested, hyphenated with gas chromatography-mass spectrometry (GC-MS), to evaluate its capability to extract volatile fractions from vegetal samples respect to classical steam distillation extraction procedure, by comparing essential oil fingerprints from three different Atractylodes species. GP-MSE showed higher extraction capabilities and sensitivities (in particular for semi-volatile components), obtaining more structured essential oil fingerprints with high reliabilities (relative standard deviation of the peak areas between 0.34% and 5.83%), requiring at the same time smaller sample amounts and reduced extraction times. Essential oil profile data were also submitted to principal component analysis and included cosine angle in order to highlight differences among the Atractylodes species under investigation. GP-MSE can be considered an alternative, simple, fast, sensitive and environmentally friendly extraction method, with high extraction efficiencies and reproducibility toward both volatile and semi-volatile compounds.

(-)-Epigallocatechin-3-gallate (EGCG) attenuates salt-induced hypertension and renal injury in Dahl salt-sensitive rats.

Epigallocatechin-3-gallate (EGCG), a main active catechin in green tea, was reported to attenuate renal injury and hypertension. However, its effects on salt-induced hypertension and renal injury remain unclear. In the present study, we explored its effects on hypertension and renal damage in Dahl rats with salt-sensitive hypertension. We found that EGCG could lower blood pressure after 6 weeks of oral administration, reduce 24 h urine protein levels and decrease creatinine clearance, and attenuate renal fibrosis, indicating that it could attenuate hypertension by protecting against renal damage. Furthermore, we studied the renal protective mechanisms of EGCG, revealing that it could lower malondialdehyde levels, reduce the numbers of infiltrated macrophages and T cells, and induce the apoptosis of NRK-49F cells. Considering that the 67 kD laminin receptor (67LR) binds to EGCG, its role in EGCG-induced fibroblast apoptosis was also investigated. The results showed that an anti-67LR antibody partially abrogated the apoptosis-inducing effects of EGCG on NRK-49F cells. In summary, EGCG may attenuate renal damage and salt-sensitive hypertension via exerting anti-oxidant, anti-inflammatory, and apoptosis-inducing effects on fibroblasts; the last effect is partially mediated by 67LR, suggesting that EGCG represents a potential strategy for treating salt-sensitive hypertension.

[One-step extraction method of phthalate esters in vegetable oils].

In order to develop a fast investigation method for phthalate esters (PAEs) from vegetable oils, a gas-liquid micro-extraction (GLME) technique that combined with GC-MS was established. A vegetable oil sample (0.1 g) was directly added into the GLME device. The integrated process of extraction, clean-up, and concentration of PAEs was completed within 5 min. Internal standard method was applied to ensure the accuracy of the results. Soybean oil, blend oil, olive oil, and sesame oil were spiked with 200 µg/kg of a mixed 15 PAEs standard, and the ranges of the recoveries and RSDs were between 60.0% to 112.3% and 0.9% to 28.4%, respectively. Compared with some traditional sample pretreatment methods such as liquid-liquid extraction, liquid-liquid micro-extraction, gel permeation chromatography, this method is simple and fast, with high accuracy, good repeatability and low matrix effect. This study verified the suitability of the GLME method for field detection of food products in food safety sector and exhibits great significance for the completion of food safety system in China.

Metabolite profiling of ginsenosides in rat plasma, urine and feces by LC-MS/MS and its application to a pharmacokinetic study after oral administration of Panax ginseng extract.

Panax ginseng is widely consumed as a functional food in the form of tea, powder, capsules, among others, and possesses a range of pharmacological activities including adaptogenic, immune-modulatory, anti-tumor, anti-aging and anti-inflammatory effects. The aim of this study was to identify and quantify the major ginsenosides and their metabolites in rat plasma, urine and feces after administration of P. ginseng extract using LC-MS/MS. We collected rat plasma samples at 0.5, 1, 2, 4, 8, 12, 24 and 48 h, and the amounts of urine and fecal samples accumulated in 24 h. Fourteen major ginsenosides and their metabolites were observed in fecal samples at high levels; however, low levels of 11 ginsenosides were detected in urine samples. The pharmacokinetics of the major ginsenosides and their metabolites was investigated in plasma. The results indicated that the maximum plasma concentration, time to maximum concentration and area under the curve of compound K were significantly greater than those of other ginsenosides. This study thus provides valuable information for drug development and clinical application of P. ginseng.

A high throughput mass spectrometry screening analysis based on two-dimensional carbon microfiber fractionation system.

A novel high-throughput, solvent saving and versatile integrated two-dimensional microscale carbon fiber/active carbon fiber system (2DµCFs) that allows a simply and rapid separation of compounds in low-polar, medium-polar and high-polar fractions, has been coupled with ambient ionization-mass spectrometry (ESI-Q-TOF-MS and ESI-QqQ-MS) for screening and quantitative analyses of real samples. 2DµCFs led to a substantial interference reduction and minimization of ionization suppression effects, thus increasing the sensitivity and the screening capabilities of the subsequent MS analysis. The method has been applied to the analysis of Schisandra Chinensis extracts, obtaining with a single injection a simultaneous determination of 33 compounds presenting different polarities, such as organic acids, lignans, and flavonoids in less than 7min, at low pressures and using small solvent amounts. The method was also validated using 10 model compounds, giving limit of detections (LODs) ranging from 0.3 to 30ngmL-1, satisfactory recoveries (from 75.8 to 93.2%) and reproducibilities (relative standard deviations, RSDs, from 1.40 to 8.06%).

Pharmacokinetics and Biodistribution of Aurantiamide and Aurantiamide Acetate in Rats after Oral Administration of Portulaca oleracea L. Extracts.

Aurantiamide and aurantiamide acetate are the main active constituents of purslane (Portulaca oleracea L.), an edible plant with various biological activities. In this study, we developed a validated UHPLC-MS/MS method to quantitate the concentrations of aurantiamide and aurantiamide acetate in the plasma and various organ tissues of rat as the basis to study their pharmacological profile and distribution in vivo. Aurantiamide and aurantiamide acetate were rapidly absorbed following oral administration, both achieving a Cmax at around 0.2 h. The extent of their metabolisms also varied among different organ tissues, resulting in about 90% reduction in concentrations 4 h after their administration, thus leaving no long-term accumulation in the tissues. This is the first study to examine the pharmacokinetic and biodistribution of aurantiamide and aurantiamide acetate in rat, and our work may serve as the first step toward the investigation of the underlying mechanisms associated with the biological activity of purslane.

Lignans and aromatic glycosides from Piper wallichii and their antithrombotic activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Piper wallichii (Miq.) Hand.-Mazz. is a medicinal plant used widely for the treatment of rheumatoid arthritis, inflammatory diseases, cerebral infarction and angina in China. Previous study showed that lignans and neolignans from Piper spp. had potential inhibitory activities on platelet aggregation. In the present study, we investigated the chemical constituents of Piper wallichii and their antithrombotic activities, to support its traditional uses. MATERIALS AND METHODS: The methanolic extract of the air-dried stems of Piper wallichii was separated and purified using various chromatographic methods, including semi-preparative HPLC. The chemical structures of the isolates were determined by detailed spectroscopic analysis, and acidic hydrolysis in case of the new glycoside 2. Determination of absolute configurations of the new compound 1 was facilitated by calculated electronic circular dichroism using time-dependent density-functional theory. All compounds were tested for their inhibitory effects on platelet aggregation induced by platelet activating factor (PAF) in rabbits׳ blood model, from which the active ones were further evaluated the in vivo antithrombotic activity in zebrafish model. RESULTS: A new neolignan, piperwalliol A (1), and four new aromatic glycosides, piperwalliosides A-D (2-5) were isolated from the stems of Piper wallichii, along with 25 known compounds, including 13 lignans, six aromatic glycosides, two phenylpropyl aldehydes, and four biphenyls. Five known compounds (6-10) showed in vitro antiplatelet aggregation activities. Among them, (-)-syringaresinol (6) was the most active compound with an IC50 value of 0.52 mM. It is noted that in zebrafish model, the known lignan 6 showed good in vivo antithrombotic effect with a value of 37% at a concentration of 30 µM, compared with the positive control aspirin with the inhibitory value of 74% at a concentration of 125µM. CONCLUSION: This study demonstrated that lignans, phenylpropanoid and biphenyl found in Piper wallichii may be responsible for antithrombotic effect of the titled plant.

[Study on sugarcane alkane alcohol to quail hyperlipidemia and atherosclerosis model].

OBJECTIVE: To study on pharmacologic actions on quail hyperlipidemia and atherosclerosis model. METHOD: To duplicate quail hyperlipidemia model by ectogenesis cholesterol and high fat forage, induce to atherosclerosis model, observe influence of sugarcane alkane alcohol to model animals' blood fat level, formation of atherosclerosis plaque, pathological changes of coronary vessels and vascular intimal. RESULT: TC, TG, LDL-C level in blood serum of quail hyperlipidemia markedly decreased after administered sugarcane alkane alcohol by dose of 30, 15, 7.5 mg x kg(-1), proliferation of aorta and brachiocephalic artery tunica intima foam cells was suppressed. CONCLUSION: Sugarcane alkane alcohol has satisfactory pharmacologic actions on hyperlipidemia and atherosclerosis animal model by regulating blood fat.

[Determination of hedychenone in Hedychium yunnanense].

OBJECTIVE: To establish an HPLC method for determination of hedychenone in Hedychium yunnanense. METHOD: C18 Chromatographic column was used, acetonitrile-water (9:1) as mobile phase, at flow rate of 1.0 mL x min(-1): The wavelength for detection was 235 nm. RESULT: The linear range of hedychenone was 5.92-29.6 microg x mL(-1)(r = 0.9999). The average recovery was 99.0%, RSD of precision was less than 2%. CONCLUSION: The method is simple, effective and feasible, and can be used to evaluate the quality of the herb.

Bergenin is the antiarrhythmic principle of Fluggea virosa.

Bergenin was isolated from the aerial parts of Fluggea virosa (Euphorbiaceae). Its structure was elucidated on the basis of chemical and spectral data. Anti-arrhythmic effects of bergenin were investigated. At concentrations of 0.2 mg/kg, 0.4 mg/kg, and 0.8 mg/kg, bergenin showed distinct therapeutic effects on BaCl2-induced arrhythmias in rats. At concentrations of 0.4 mg/kg and 0.8 mg/kg bergenin significantly countered arrhythmias induced by ligation and reperfusion of the coronary artery. At 0.8 mg/kg, bergenin elevated the atria fibrillation threshold in rabbits from 1.34 mV to 1.92 mV. Our results suggest that bergenin has good potential to treat cardiac arrhythmias.