Detection and Characterization of the Metabolites of Ciwujianoside B in Rats Based on UPLC-Fusion Lumos Orbitrap Mass Spectrometry.

We previously conducted a systematic study on the metabolic process and products of hederasaponin B in rats. We hypothesized that the sugar chain structures play a key role in the metabolism of triterpenoid saponins. To verify this hypothesis, we conducted metabolic research on ciwujianoside B ascribed to the same sugar chains and a distinct aglycone and compared it with hederasaponin B. Specifically, we collected feces, urine, and plasma of rats after gavage with ciwujianoside B and identified 42 metabolites by UPLC-Fusion Lumos Orbitrap mass spectrometry. Finally, ciwujianoside B metabolism and hederasaponin B metabolism were compared, reaching the following conclusions: (i) more than 40 metabolites were identified in both, with the majority of metabolites identified in feces; (ii) the corresponding metabolic pathways in vivo were basically similar, including deglycosylation, acetylation, hydroxylation, glucuronidation, oxidation, and glycosylation; and (iii) deglycosylation was considered the main metabolic reaction, and its metabolites accounted for approximately 50% of all metabolites. Overall, this study provides a foundation for further research on the metabolism of triterpenoid saponins.

A metabolism-based study of the mechanism of action of Scrophularia ningpoensis Hemsl. on nephrogenic edema.

Nephrogenic edema (NE) is a type of edema with hypoproteinemia and water and sodium retention as a result of renal injury. Traditional Chinese medicine has proved that Scrophularia ningpoensis Hemsl. has an effect on NE, but its mechanism is not clear. In this study, the main components and blood components of S. ningpoensis were identified using ultra-high-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS). Pathological section and blood biochemical analysis were used to estimate the therapeutic effect of S. ningpoensis on NE. Network pharmacology was used to predict the potential pathways of S. ningpoensis. The metabolomics method was used to study the changes in small-molecule metabolites in the body. The results showed that S. ningpoensis could relieve NE by regulating relative to renal function and body edema, and its mechanism may be related to the regulation of energy metabolism, recovery of renal injury, and reduction in inflammation. The active component harpagoside may be one of the important compounds of S. ningpoensis in the treatment of NE. We confirmed that S. ningpoensis has a therapeutic effect on NE, which provides a solid scientific research basis for the clinical application of S. ningpoensis.

The association between neuropeptide oxytocin and neuropsychiatric disorders after orthopedic surgery stress in older patients.

BACKGROUND: The health outcomes of geriatric patients exposed to surgery were found to be enhanced by social support and stress management. The aim of this study was to characterise the relationship between oxytocin and neuropsychiatric disorders after surgery. METHODS: A total of 132 geriatric patients aged ≥ 60 years received orthopedic surgery in the First Affiliated Hospital of Harbin Medical University (Harbin, China) were enrolled in the present study. The salivary levels of stress hormone cortisol and oxytocin were measured by enzyme-linked immunosorbent assay for the screening of the stress state and oxytocin function. Moreover, the Depression Anxiety and Stress Scale (DASS), the Geriatric Anxiety Inventory (GAI), the Geriatric Depression Scale (GDS) and the Montgomery-Åsberg Depression Rating Scale (MADRS) were conducted to identify the severity of anxiety and depression. The association between oxytocin and mental health was performed by linear regression analyses in older patients receiving orthopedic surgery. Finally, the Duke Social Support Index (DSSI) was selected to measure the social support and the potential link to mental outcomes. RESULTS: The scores from questionnaires showed that female patients with higher social support and higher levels of oxytocin demonstrated better stress-reducing responses as reflected by lower cortisol and decreased anxiety and depression symptoms. Regression analyses revealed that there was a significant association between oxytocin and scores in DASS, GAI, GDS, MADRS and DSSI, suggesting a potential link between peripheral oxytocin function and mood outcomes after orthopedic surgery. CONCLUSIONS: Our findings reveal that oxytocin enhances the stress-protective effects of social support and reduces anxiety and depression states under stressful circumstances, particularly in older women receiving orthopedic surgery.

[Mechanism of Huanglian Wendan Decoction in improving impaired glucose tolerance based on skeletal muscle NLRP3/caspase-1/IL-1ß, IL-18 pathway].

This study investigated the mechanism of improving impaired glucose tolerance(IGT) of rats by Huanglian Wendan Decoction from the perspective of the skeletal muscle Nod-like receptor protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/interleukin-1ß(IL-1ß), interleukin-18(IL-18) pathway. Healthy male SD rats were fed with the diet containing 45% fat for 20 weeks, accompanied by a high-temperature and high-humidity environment and an inactive lifestyle, for the establishment of the IGT rat model. The rats were divided into the blank control group, model control group, metformin hydrochloride group(positive drug group, 0.05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group(7.8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks, the obesity and glycemic indexes of all the rats were measured. The fasting serum insulin(FINS) level was determined by ELISA, with the insulin sensitivity index(ISI) and insulin resistance index(IRI) calculated. The mRNA and protein expression le-vels of nuclear factor kappaB(NF-κB), NLRP3, caspase-1, IL-1ß and IL-18 in skeletal muscle tissue were detected by real-time polymerase chain reaction(PCR), Western blot and immunofluorescence. Compared with the blank control group, the model control group witnessed significantly increased mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1ß and IL-18. As revealed by the comparison with the model control group, Huanglian Wendan Decoction could effectively regulate the obesity status, reduce body weight, correct the abnormal levels of 2-hour plasma glucose(2 hPG), insulin resistance index(IRI), insulin sensitivity index(ISI), and lower the mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1ß and IL-18 in the skeletal muscle tissue of IGT rats. Combined with previous studies, the above results showed that the occurrence and development of IGT was closely related to inflammatory response and the classic pyroptosis pathway in skeletal muscle, such as NLRP3/caspase-1/IL-1ß, IL-18. It is inferred that the mechanism of Huanglian Wendan Decoction was to alleviate insulin resistance(IR) and then reverse the course of IGT lies in the regulation of the abnormal insulin receptor signaling pathway based on the NLRP3 inflammasome pathway.

Chronic administration of ellagic acid improved the cognition in middle-aged overweight men.

This study aimed to investigate if ellagic acid has beneficial effects on cognitive deficits in middle-aged overweight individuals and to propose a possible mechanism. A total of 150 middle-aged male participants, including 76 normal-weight and 74 overweight men, aged between 45 to 55 years, were recruited for this study. Both normal-weight and overweight participants were administered either 50 mg ellagic acid or placebo cellulose daily for 12 weeks. Blood lipids, peripheral brain-derived neurotrophic factor (BDNF), and saliva cortisol were assessed on the last day of the procedure to investigate the effects induced by ellagic acid. The results revealed that ellagic acid treatment improved the levels of blood lipid metabolism with a 4.7% decline in total cholesterol, 7.3% decline in triglycerides, 26.5% increase in high-density lipoprotein, and 6.5% decline in low-density lipoprotein. Additionally, ellagic acid increased plasma BDNF by 21.2% in the overweight group and showed no effects on normal-weight participants. Moreover, the increased saliva cortisol level in overweight individuals was inhibited by 22.7% in a 12-week ellagic acid treatment. Also, compared with placebo, overweight individuals who consumed ellagic acid showed enhanced cognitive function as measured by the Wechsler Adult Intelligence Scale-Revised and the Montreal Cognitive Assessment. To the best of our knowledge, this is the first report showing that ellagic acid prevents cognitive deficits through normalization of lipid metabolism, increase in plasma BDNF level, and reduction of saliva cortisol concentration. These results indicate that ellagic acid has a potential to restore cognitive performance related to mild age-related declines.

[Effects of Scrophulariae Radix and Split Component on Isoproterenol-Induced Ventricular Remodeling in Rat].

Objective: To explore the effects and mechanism of Scrophulariae Radix and split component on experimental ventricular remodeling. Methods: The rats were injected subcutaneously by 10,5 mg / kg ISO for 2 d and then 3 mg / kg ISO as maintenance dose for 7 d to build the the ventricular remodeling. After 21 days' treatment,the left ventricular mass index( LVMI) and heart mass index( HMI) were measured. The content of atrial natriuretic peptide( ANP),endothelin-1( ET-1) and angiotensinⅡ( Ang II) were determined by enzyme linked immunosorbent assay( ELISA),and the pathological changes of myocardial tissue were also observed. Results: LVMI,HMI were improved by total composition components and small polar iridoid glycosides components,and the content of ANP,ET-1 and AngⅡwas decreased remarkably; The polysaccharide components could only decline the content of Ang Ⅱ and ET-1. Conclusion: The pharmacological effects of Scrophulariae Radix inhibit ventricular remodeling may be related to the small polar iridoid glycosides components and the polysaccharide components. And the mechanism may be related to regulating the over expression neurohumor factors and inhibiting the release of ANP and ET-1,Ang Ⅱ.

[Progress in the molecular genetic mechanism of gonadoblastoma].

Gonadoblastoma (GB), a rare in situ germ cell tumor derived from sex cord and germ cells, is closely associated with gonadal dysgenesis. About 80% of GB individuals exhibit 46, XY female phenotype while the others are 45, XY and 46, XX with disorders of sex development. Moreover, 35% of GB can eventually develop into malignant tumors, such as seminoma and dysgerminoma tumors. The molecular genetic mechanism of GB remains to be fully uncovered due to phenotypic and genetic heterogeneity. Increasing studies show that the formation of GB is closely related to genes regulating sexual differentiation and determination (e.g., SRY, WT1, SOX9, Foxl2, TSPY, etc), and is affected by the interaction of genetic and epigenetic regulation. Here we describe the clinical and pathological features, diagnosis and treatment of GB, and also summarize the molecular genetic and epigenetic mechanisms underlying the gonadal abnormalities that lead to GB. We analyze and construct the common gene regulatory networks related to the development of GB, and describe some obstacles and deficiencies in current studies to provide innovative perspectives on further studying the pathological and molecular mechanisms of GB.

An improved acute gouty arthritis rat model and therapeutic effect of rhizoma Dioscoreae nipponicae on acute gouty arthritis based on the protein-chip methods.

Rhizoma Dioscoreae nipponicae (RDN) is an herbal medicine. In the theories of Traditional Chinese Medicine (TCM), the function of RDN is to expel wind and remove dampness. Inflammatory mechanisms play an important role in the pathological process and prognosis of acute gouty arthritis (AGA). The aim of this study was to determine the specially expressed proteins through testing the proteins of the synovium in rats with AGA. The animal model of AGA was set up by Monosodium urate crystal (MSU) combined with hypoxanthine (HX), which was ameliorated in our previous experiment. Blood samples for measurement of serum uric acid were collected prior to sacrifice. Outcomes were assessed (two days after injection) by histological stain and protein quantitation. Three chips of RayBio® Human Label-based Antibody Array I were applied to detect 90 proteins in the synovium tissue of AGA rats. 14 differently expressed proteins were found in the synovium of AGA rats, and nine of them were first found in this model. There were seven up-regulated and seven down-regulated proteins, both TRAIL and Neuropilin-2 could be identified as key contributors to the pathomechanism of AGA.

Metabolomic study of a rat fever model induced with 2,4-dinitrophenol and the therapeutic effects of a crude drug derived from Coptis chinensis.

This study describes the metabonomics of fevers in animal models and the therapeutic effects of Rhizoma coptidis extract (RCE) on them. The rat urinary samples were analyzed by UPLC/ ESI-Q-TOF/MS, combined with principal component analysis (PCA). Nine ions were chosen to characterize the similarities and differences in the responses to fever. The ion at m/z 206.0278 was unambiguously identified to be xanthurenic acid. This study demonstrated that the metabonomic approach can readily distinguish between febrile and healthy individuals. This data support the contention that the metabonomic approach represents a promising new technology for the development of rapid-throughput in vivo fever screening. Furthermore, this approach can detect the interfering effects of RCE. This investigation has led the authors to believe that metabonomics is a valid approach for explaining the therapeutic effects of traditional Chinese medicine on fevers.

Protective effect of effective part of Acanthopanacis senticosus on damage of PC12 cells induced by MPP+.

The purpose of our research was to evaluate the protective effect of the effective part of Acanthopanacis senticosus (AS) on the damage of PC12 cells induced by MPP+, an in vitro cell model for Parkinson's disease. Cell viability and apoptosis of PC12 cells induced by MPP' were assayed by MTT and flow cytometry respectively in the presence or absence of the effective part of AS. The contents of lactate dehydrogenase (LDH), nitric oxide (NO), nitric oxide synthase (NOS) and malondialdehyde (MDA) were determined by UV spectrophotometer. Our study showed that the survival rate of PC12 cells was markedly increased while cell apoptosis was decreased in the range of 200 to 400 mg x L(-1) of the effective part of AS. The contents of LDH, NO, NOS, MDA were reduced. Our experimental results indicated that the effective part of AS had the protective effect on the damage of PC12 cells induced by MPP+. The underlying mechanisms might be the combination of reduction of LDH leakage and decreases in the contents of NO, NOS and MDA, and finally prevent the apoptosis in PC12 cells and increase the cell survival rate.