TOM40 mediates the effect of TSPO on postpartum depression partially through regulating calcium homeostasis in microglia.

AIMS: To assess the effect of the translocator protein 18 kDa (TSPO) on postpartum depression and explore its mechanism. METHODS: Postpartum depression (PPD) mouse model was established, and flow cytometry, immunofluorescence, Western blot analysis, real-time quantitative PCR, adeno-associated virus (AAV), co-immunoprecipitation-mass spectrometry and immunofluorescence co-staining were used to detect the effect of TSPO ligand ZBD-2 on PPD mice. RESULTS: ZBD-2 inhibits the overactivation of microglia in the hippocampus and amygdala of PPD model mice. ZBD-2 not only inhibited the inflammation but also repressed the burst of reactive oxygen species (ROS) and mitochondrial ROS (mtROS). Meanwhile, ZBD-2 protects mitochondria from LPS-induced damages through inhibiting the influx of calcium. ZBD-2 modulated the calcium influx by increasing the level of translocase of the outer mitochondrial membrane 40 (TOM40) and reducing the interaction of TSPO and TOM40. In addition, the effect of ZBD-2 was partially dependent on anti-oxidative process. Knockdown of TOM40 by adeno-associated virus (AAV) in the hippocampus or amygdala dramatically reduced the effect of ZBD-2 on PPD, indicating that TOM40 mediates the effect of ZBD-2 on PPD. CONCLUSIONS: TOM40 is required for the effect of ZBD-2 on treating anxiety and depression in PPD mice. This study reveals the role of microglia TSPO in PPD development and provides the new therapeutic strategy for PPD.

PJA1 mediates the effects of astrocytic GPR30 on learning and memory in female mice.

Hormone replacement therapy (HRT) is not recommended for treating learning and memory decline in menopausal women because it exerts adverse effects by activating classic estrogen receptors ERα and ERß. The membrane estrogen receptor G protein-coupled receptor 30 (GPR30) has been reported to be involved in memory modulation; however, the underlying mechanisms are poorly understood. Here, we found that GPR30 deletion in astrocytes, but not in neurons, impaired learning and memory in female mice. Astrocytic GPR30 depletion induced A1 phenotype transition, impairing neuronal function. Further exploration revealed that Praja1 (PJA1), a RING ubiquitin ligase, mediated the effects of astrocytic GPR30 on learning and memory by binding to Serpina3n, which is a molecular marker of neuroinflammation in astrocytes. GPR30 positively modulated PJA1 expression through the CREB signaling pathway in cultured murine and human astrocytes. Additionally, the mRNA levels of GPR30 and PJA1 were reduced in exosomes isolated from postmenopausal women while Serpina3n levels were increased in the plasma. Together, our findings suggest a key role for astrocytic GPR30 in the learning and memory abilities of female mice and identify GPR30/PJA1/Serpina3n as potential therapeutic targets for learning and memory loss in peri- and postmenopausal women.

Evaluation of Radio Frequency-Assisted Enzymatic Extraction of Non-Anthocyanin Polyphenols from Akebia trifoliata Flowers and Their Biological Activities Using UPLC-PDA-TOF-ESI-MS and Chemometrics.

A new radio frequency heating-assisted enzymatic extraction (RF-E) method is applied for the determination of phenolic compounds in Akebia trifoliata flowers, compared with hot water, acidified ethanol (EtOH), and enzymatic-assisted (EA) extractions. Non-anthocyanin polyphenol profiles, antibacterial, angiotensin-converting enzyme (ACE) inhibitory, anti-inflammatory activities, and structures of extracts are evaluated. Results show no significant differences in the extraction of total flavonoid content (15.85-16.63 mg QEs/g) and ACE inhibitory activity (51.30-52.86%) between RF-E and EA extracts. RF-E extract shows the highest anti-inflammatory activities. FTIR and UV spectra reveal that acidified EtOH treatment has a significant effect on the structure of the extract due to its highest flavonoid content (20.33 mg QEs/g), thus it has the highest antibacterial activity against Staphylococcus aureus and Escherichia coli. Sixteen non-anthocyanin polyphenols are identified by UPLC-PDA-TOF-ESI-MS and RF pre-treatment did not cause significant compound degradation. The chemometric analysis shows that enzymatic hydrolysis significantly increased biological activities, and the presence of non-anthocyanin polyphenols correlates well with ACE inhibitory and anti-inflammatory activities. Accordingly, A trifoliata flowers have potential as reagents for the food and pharmaceutical industries due to their abundant polyphenols that could be extracted efficiently using RF-E.

Effect of radio frequency, ultrasound, microwave-assisted papain, and alcalase hydrolysis on the structure, antioxidant activity, and peptidomic profile of Rosa roxburghii Tratt. seed protein.

Traditional enzymatic hydrolysis methods have defects such as low efficiency and poor bioactivity in the production of active peptides. In this study, radio frequency (RF) technology was innovatively used to assist the hydrolysis of Rosa roxburghii Tratt. seed protein (RTSP) by papain and alcalase. RF-assisted hydrolysis was compared with ultrasound-(US) and microwave (MW)-assisted techniques in terms of the degree of hydrolysis, structure, antioxidant properties, and changes in the peptidome of the hydrolysates to clarify the mechanism of functional change of physically-assisted hydrolysate. All three methods improved hydrolysis efficiency. The degree of hydrolysis (DH) of papain group increased from 6.38% to 7.97%, 9.97% and 8.37% after US-, MW- and RF-assisted hydrolysis, respectively, while the DH of alcalase-treated group increased from 21.13% to 25.66%, 26.03%, and 23.01%, respectively. The in vitro antioxidant capacity and intracellular antioxidant capacity of RTSP and its hydrolysates were measured and evaluated by fuzzy statistical evaluation, and MW-assisted alcalase hydrolysis had the highest in vitro and intracellular antioxidant activity scores of 0.713 and 0.820, respectively. Fourier transform infrared and amino acid composition analysis explained the enhanced antioxidant properties of the hydrolysates. Further peptide profiling showed the physical assistance led to an increase in the species and contents of small molecule antioxidant peptides compared to enzyme treatment alone. Pearson's linear correlation analysis showed that AY, LY, IY, PHW, SVL, LHL, YYV, VYY, and NHAV were significantly correlated with the antioxidant properties of hydrolysates. Our data suggested that physical assistance such as US, MW, and RF were effective to improve the efficiency of enzymatic hydrolysis and produce novel antioxidant peptides. PRACTICAL APPLICATION: In this study, it was found that electromagnetic wave-assisted enzymatic hydrolysis could improve the efficiency of hydrolysis and enhance the antioxidant activity of hydrolysates compared to unassisted means. Compared with MW treatment, RF has the comparable hydrolysis effect, but has the advantages of high penetration ability, good uniformity, and low energy consumption and has greater potential for the production of bioactive peptides.

Activation of GIPR Exerts Analgesic and Anxiolytic-Like Effects in the Anterior Cingulate Cortex of Mice.

Background: Chronic pain is defined as pain that persists typically for a period of over six months. Chronic pain is often accompanied by an anxiety disorder, and these two tend to exacerbate each other. This can make the treatment of these conditions more difficult. Glucose-dependent insulinotropic polypeptide (GIP) is a member of the incretin hormone family and plays a critical role in glucose metabolism. Previous research has demonstrated the multiple roles of GIP in both physiological and pathological processes. In the central nervous system (CNS), studies of GIP are mainly focused on neurodegenerative diseases; hence, little is known about the functions of GIP in chronic pain and pain-related anxiety disorders. Methods: The chronic inflammatory pain model was established by hind paw injection with complete Freund's adjuvant (CFA) in C57BL/6 mice. GIP receptor (GIPR) agonist (D-Ala2-GIP) and antagonist (Pro3-GIP) were given by intraperitoneal injection or anterior cingulate cortex (ACC) local microinjection. Von Frey filaments and radiant heat were employed to assess the mechanical and thermal hypersensitivity. Anxiety-like behaviors were detected by open field and elevated plus maze tests. The underlying mechanisms in the peripheral nervous system and CNS were explored by GIPR shRNA knockdown in the ACC, enzyme-linked immunosorbent assay, western blot analysis, whole-cell patch-clamp recording, immunofluorescence staining and quantitative real-time PCR. Results: In the present study, we found that hind paw injection with CFA induced pain sensitization and anxiety-like behaviors in mice. The expression of GIPR in the ACC was significantly higher in CFA-injected mice. D-Ala2-GIP administration by intraperitoneal or ACC local microinjection produced analgesic and anxiolytic effects; these were blocked by Pro3-GIP and GIPR shRNA knockdown in the ACC. Activation of GIPR inhibited neuroinflammation and activation of microglia, reversed the upregulation of NMDA and AMPA receptors, and suppressed the enhancement of excitatory neurotransmission in the ACC of model mice. Conclusions: GIPR activation was found to produce analgesic and anxiolytic effects, which were partially due to attenuation of neuroinflammation and inhibition of excitatory transmission in the ACC. GIPR may be a suitable target for treatment of chronic inflammatory pain and pain-related anxiety.

Antimicrobial, antioxidant and physical properties of chitosan film containing Akebia trifoliata (Thunb.) Koidz. peel extract/montmorillonite and its application.

A novel active packaging film was prepared in this study that incorporated Akebia trifoliata (Thunb.) Koidz. peel extracts (APE) and montmorillonite (MMT) into chitosan (CH) films. Compared with the pure CH film, the CH/APE film showed significantly higher tensile strength, elongation at break, UV light resistance, and antibacterial activity; the CH/MMT film displayed significant increases in contact angle, antioxidant activity, oxygen permeability, and thermal stability. SEM and AFM analyses showed that the additions were well-distributed into the CH matrix, but MMT induced a more compact and rougher structure. The CH-based film formula was optimized using the single-factor test and Box-Behnken design and was 0.15% MMT, 0.15% APE, and 1.50% CH. Besides, the optimized coating was applied in the postharvest preservation of A. trifoliata fruits, which yielded a significant effect on the delaying crack and mature of the fruits during 35 days of storage at 5 °C.

Structure, physicochemical and bioactive properties of dietary fibers from Akebia trifoliata (Thunb.) Koidz. seeds using ultrasonication/shear emulsifying/microwave-assisted enzymatic extraction.

Dietary fiber extracts from Akebia trifoliata (Thunb.) Koidz. seeds were generated using alkaline extraction, enzymatic hydrolysis as well as physical pretreatments including ultrasonication, shear emulsifying and microwave. Compared with the basic alkaline hydrolysis, cellulase-assisted extraction exhibited the highest water holding capacity (7.99 g/g), cation exchange capacity (0.55 mmol/g), glucose adsorption capacity (1352.33 µmol/g) and pancreatic lipase inhibition (37.69%). Ultrasonication pretreatment before both the preceding procedures increased the oil holding capacity (3.95 g/g) and α-amylase inhibition (6.81%) of the extracted fiber. Microwave pretreatment generated fiber possessing the greatest specific surface area (137.70 m2/kg), water swelling capacity (1.25 mL/g), adsorption capacity (4.14 mg/g, pH = 2) and sodium cholate adsorption capacity (38.68%). However, shear emulsifying gave the lowest crude yields (57.72%), glucose absorption capacity (22.09 mg/g), α-amylase inhibition (2.77%) and pancreatic lipase inhibition (22.61%) though it contained highest levels of soluble fiber (6.40 g/100 g) and lowest crystallinity (33.6%). A fuzzy statistical evaluation indicated that ultrasonication assisted enzymatic hydrolysis was the most efficient improvement in the overall properties of the dietary fiber. These results provide valuable information for the potential use of A. trifoliata seeds as a new food ingredient and for extraction of dietary fiber from this plant.

An Epileptic Patient with Recurrent Hyperbilirubinemia Caused by Gilbert Syndrome.

Gilbert syndrome (GS) is characterized by intermittent indirect bilirubin elevation. Several antiepileptic drugs (AEDs) impair the liver function to different degrees, such as valproic acid, lamotrigine, phenobarbital, phenytoin, and carbamazepine. Herein, we present the case of a 26-year-old epileptic patient with frequently recurring mild hyperbilirubinemia during taking AEDs. After repeated adjustment of the doses and types of AEDs, the bilirubin level still remained elevated. He was then referred to the Gastroenterology Department. The results of diagnostic tests, clinical manifestation, imaging studies, liver biopsy and whole-exome sequencing all made contributions to our conclusion that GS played an important role in the elevation of bilirubin. Ultimately, his seizure was controlled by levetiracetam (500 mg per day) and he was advised to periodically undergo the liver function tests.

TGF-ß Signaling Promotes Glioma Progression Through Stabilizing Sox9.

Gliomas are brain and spinal cord malignancies characterized by high malignancy, high recurrence and poor prognosis, the underlying mechanisms of which remain largely elusive. Here, we found that the Sry-related high mobility group box (Sox) family transcription factor, Sox9, was upregulated and correlated with poor prognosis of clinical gliomas. Sox9 promotes migration and invasion of glioma cells and in vivo development of xenograft tumors from inoculated glioma cells. Sox9 functions downstream of the transforming growth factor-ß (TGF-ß) pathway, in which TGF-ß signaling prevent proteasomal degradation of the Sox9 protein in glioma cells. These findings provide novel insight into the wide interplay between TGF-ß signaling and oncogenic transcription factors, and have implications for targeted therapy and prognostic assessment of gliomas.

Diagnostic Value of the Survivin Autoantibody in Four Types of Malignancies.

BACKGROUND: Tumor-associated antigen overexpression, which has been reported in many types of cancers, may trigger autoantibody secretion. The present study was designed to test whether levels of circulating autoantibodies to survivin protein-derived antigens is altered in liver, esophageal, breast, and lung cancers. METHODS: Patients with liver (144), esophageal (159), breast (124), and lung cancers (267), and healthy volunteers (362) were recruited for the study, and serum samples were collected for ELISA autoantibody analysis. RESULTS: Compared with the control group, survivin autoantibody levels were significantly higher in serum from patients with breast cancer and lung cancer, but were significantly lower in serum from patients with liver cancer (p < 0.05). In stage I and II lung cancer, the best-fit areas under the receiver operating characteristic curve was 0.731 (standard error [SE] = 0.023; 95% confidence interval [CI] 0.687-0.776) and the sensitivity, with 90% specificity, was 23.7%. CONCLUSION: Analysis across four types of malignancies revealed that the survivin autoantibody had good specificity and sensitivity in lung cancer. Circulating autoantibodies to survivin could be a potential biomarker for the early lung cancer diagnosis.

Synthesis and evaluation of trehalose-based compounds as novel inhibitors of cancer cell migration and invasion.

As a continuous research for the discovery of trehalose-based anti-invasive agents, we developed a convenient synthetic approach for the preparation of 6,6'-dideoxy-6,6'-bis(acylamino)-α,α-D-trehaloses. A series of trehalose-based amides were prepared through the trityl protection of the two primary hydroxyls of α,α-D-trehalose, benzoylation, the removal of the trityl protective group, mesylation, azidation, catalytic hydrogenation in the presence of hydrochloride, coupling reaction with a variety of acids, and subsequent debenzoylation and deacetylation in some cases. Compound 8b, 6,6'-dideoxy-6,6'-bis(2-hydroxybenzamide)-α,α-D-trehalose, was just as potent as the natural brartemicin against the invasion of murine colon 26-L5 cells. It exhibited no cytotoxicity on human breast adenocarcinoma MDA-MB-231 and murine colon 26-L5 cells. It can significantly inhibit the migration and invasion of the MDA-MB-231 cells. The anti-invasive effect of 8b was possibly related to its inhibitory activity on MMP-9, its suppression on the expression of MMP-9 and VEGF, and its deactivation of Akt.

Synthesis and structure-activity relationships studies of brartemicin analogs as anti-invasive agents.

Brartemicin is a trehalose-based inhibitor of tumor cell invasion produced by the actinomycete of the genus Nonomuraea. In order to find more potent anti-invasive agents and study the structure-activity relationships, a series of 19 brartemicin analogs were prepared via two synthetic routes from α,α-D-trehalose and evaluated for their anti-invasive activities. Compound 4f, 6,6'-bis(2,3-dimethoxybenzoyl)-α,α-D-trehalose, was more potent than the natural brartemicin. It inhibited the invasion of murine colon 26-L5, colon carcinoma SW620, melanoma B16-BL6 and breast MDA-MB-231 cells with IC50 values of 0.15, 2.35, 4.12 and 2.61 µM, respectively. Analog 4p, 6,6'-bis(3,4-dimethoxycinnamoyl)-α,α-D-trehalose, was as potent as brartemicin against invasion of murine colon 26-L5 carcinoma cells in vitro. The structure-activity relationships of these novel trehalose-based compounds were summarized.

Synthesis and evaluation of trehalose-based compounds as anti-invasive agents.

Brartemicin is a trehalose-based inhibitor of tumor cell invasion produced by the actinomycete of the genus Nonomuraea. In order to explore the preliminary structure-activity relationship and obtain more potent inhibitors, a series of brartemicin analogs were synthesized through the Mitsunobu coupling of the secondary hydroxyls benzyl protected α,α-D-trehalose with benzoic acid derivatives, followed by modification of functional groups and deprotection. These compounds were evaluated for their inhibitory activity against invasion of murine colon 26-L5 carcinoma cells in vitro. Among the synthetic analogs tested, 6,6'-bis(2,3-dimethoxybenzoyl)-α,α-D-trehalose (5e) was found to be the most potent anti-invasive agent, exhibited a 2.6-fold improvement with regard to the parent natural product brartemicin, and it is considered to be a promising lead molecule for the anti-metastasis.