Autophagy inhibition mediated by trillin promotes apoptosis in hepatocellular carcinoma cells via activation of mTOR/STAT3 signaling.

Apoptosis and autophagy have been shown to act cooperatively and antagonistically in self-elimination process. On the one side, apoptosis and autophagy can act as partners to induce cell death in a coordinated or cooperative manner; on the flip side, autophagy acts as an antagonist to block apoptotic cell death by promoting cell survival. Our previous research indicated that trillin could induce apoptosis of PLC/PRF/5 cells, but the effects of trillin on autophagy as well as its functional relationship to apoptosis have not been elucidated. Here, the running study aims to investigate the function and molecular mechanism of trillin on autophagy with hepatocellular carcinoma (HCC) cells. The objective of this study is to investigate the molecular mechanism of trillin on autophagy in HCC cells. Protein levels of autophagy markers beclin1, LC3B, and p62 were detected by western blotting. 6-Hydroxyflavone and stattic were used to test the role of trillin regulation of autophagy via serine threonine kinase (AKT)/extracellular-regulated protein kinases (ERK) 1/2/mammalian target of rapamycin (mTOR)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Flow cytometry was used to detect caspase 3 activity and apoptosis in PLC/PRF/5 cells treated with trillin for 24 h with or without rapamycin, stattic, and 6-hydroxyflavone. The protein level of autophagy marker beclin1 was decreased, whilst the protein level of p62 was significantly increased by trillin treatment, indicating trillin treatment led to inhibition of autophagy in HCC cells. Trillin treatment could reduce the protein levels of p-AKT and p-ERK1/2, but enhance the protein levels of mTOR and p-mTOR, suggesting that trillin could inhibit AKT/ERK rather than mTOR. The AKT/ERK activator 6-hydroxyflavone could reverse the loss of AKT and ERK1/2 phosphorylation induced by trillin, implying that trillin impairs autophagy through activated mTOR rather than AKT/ERK. STAT3 and p-STAT3 were significantly upregulated by the trillin treatment with an increase in dose from 0 to 50 µM, suggesting that autophagy inhibition is mediated by trillin via activation of STAT3 signaling. The STAT3 inhibitor stattic significantly reversed the increased STAT3 phosphorylation at tyrosine 705 induced by trillin. The mTOR signaling inhibitor rapamycin reversed the trillin-induced mTOR phosphorylation enhancement but exerted no effects on total mTOR levels, suggesting trillin treatment led to inhibition of autophagy in HCC cells through activating mTOR/STAT3 pathway. Furthermore, caspase 3 activities and the total rate of apoptosis were increased by trillin treatment, which was reversed by rapamycin, stattic, and 6-hydroxyflavone, proving that trillin promotes apoptosis via activation of mTOR/STAT3 signaling. Trillin induced autophagy inhibition and promoted apoptosis in PLC/PRF/5 cells via the activation of mTOR/STAT3 signaling. Trillin has the potential to be a viable therapeutic option for HCC treatment.

Modulating Oxygen Vacancies in Lead Chromate for Photoelectrocatalytic Water Splitting.

Although modulating oxygen vacancies in semiconductors has attracted broad interest in photocatalysis and photoelectrocatalysis, identifying the intrinsic roles of oxygen vacancies on photoelectrocatalytic properties is often elusive. In this work, the oxygen vacancies in a typical semiconductor lead chromate (PbCrO4 ) are regulated via controlling the oxygen chemical potentials of O-poor and O-rich post-annealing atmospheres. Oxygen vacancies identified in PbCrO4 can introduce electronically shallow energy levels and deep energy levels owing to the symmetry difference of oxygen atoms in the structure. A higher population of deep energy levels created under O-poor atmosphere induces the formation of more surface trapped states, resulting in a higher photovoltage for charge separation. Meanwhile, the existence of surface trapped states can significantly improve the charge injection efficiency of the PbCrO4 photoanode and enhance the water oxidation activity. By modulating oxygen vacancies in the PbCrO4 photoanode, a photocurrent density of 3.43 mA cm-2 at 1.23 V vs reversible hydrogen electrode (RHE) under simulated AM1.5G is acheived. Further passivation of surface trapped states and introducing the water oxidation cocatalyst CoPi lead to a record applied bias photon-to-current efficiency (ABPE) of 1.12%. This work provides a guide to understand the mechanism of oxygen vacancies in oxide-based semiconductor photocatalysis and photoelectrocatalysis.

Boosting Photocatalytic Water Oxidation on Photocatalysts with Ferroelectric Single Domains.

Ferroelectric materials are considered as promising photocatalysts due to their efficient charge separation via a polarization-induced built-in electric field. However, the polydomain structures hinder spatial charge separation and transfer due to the cancellation of polarization vectors in the domains. In this work, taking BiFeO3 (BFO) as a prototype, single-domain BFO nanosheets with visible-light absorption are prepared, as evident by piezoresponse force microscopy (PFM), spatially resolved surface photovoltage spectroscopy (SRSPS), and photodeposition experiments. The single-domain BFO nanosheets show nine times activity in photocatalytic water oxidation reaction under visible-light irradiation, compared with that of the polydomain BFO particles. With the asymmetric driving force for charge separation in a single domain, selective deposition of cocatalysts further enhances the photocatalytic activity of single-domain ferroelectric BFO nanosheets. These results demonstrate the role of the single-domain structure in constructing the driving force of charge separation in ferroelectric photocatalysts. The fabrication of single-domain structures in ferroelectric photocatalysts to achieve enhanced photocatalytic activity offers a path to efficiently utilize the photogenerated charges in solar energy conversion.

Charge Separation between Pt Co-catalysts and Plasmonic Au in Pt-Au/C3N4 Photocatalysts.

Chemical processes induced by surface plasmon resonance have received great attention due to the wide spectral absorption and tunable optical property. Co-catalysts have been introduced into plasmonic-metal/semiconductor photocatalysts to inhibit the recombination of plasmon-induced carriers. However, it is unclear how the locations of co-catalysts (on the surface of plasmonic metal or the semiconductor) affect the plasmonic photocatalystic reactions. Herein, we report that Pt co-catalysts can be selectively deposited on Au nanoparticles (NPs) of Au/C3N4 photocatalysts through an atomic layer deposition method. Compared with the Pt co-catalysts exclusively on C3N4, Pt solely on Au NPs has a 4.5-fold increase in activity for plasmonic hydrogen evolution. The reduced photoluminescence intensity and prolonged photoluminescence lifetime reveal that Pt solely on Au NPs provides higher charge separation efficiency. The enhanced photocatalytic activity of Pt co-catalysts solely on Au NPs is attributable to the more efficient and direct utilization of the plasmon resonance-induced electrons and separation of electrons and holes.

Internal-Field-Enhanced Charge Separation in a Single-Domain Ferroelectric PbTiO3 Photocatalyst.

Ferroelectric materials with spontaneous polarization-induced internal electric fields have drawn increasing attention in solar fuel production due to the intrinsic polarized structure. However, the origination of charge separation in these materials at the nano/microlevel is ambiguous owing to the complexity of the multielectric fields. Besides, the observed charge separation ability is far from theoretical expectation. Herein, by spatially resolved surface photovoltage spectroscopy, it is clearly demonstrated that the depolarization field in single-domain ferroelectric PbTiO3 (PTO) nanoplates is the main driving force for charge separation and it can effectively drive photogenerated electrons and holes to the positive and negative polarization facets, respectively. Moreover, the charge separation ability of PTO nanoplates increases with increasing particle size along the polarization direction, due to the increasing potential difference between the opposite polarization facets. Furthermore, this driving force for charge separation directly contributes to the enhancement of the photocatalytic hydrogen evolution reaction activity in ferroelectrics. Finally, it is proved that the screening field compensates part of the depolarization field and can be diminished by adding a dielectric layer on the ferroelectric surface. These findings demonstrate the importance of increasing the depolarization field and decreasing the screening field for efficient charge separation in ferroelectric semiconductor photocatalysts.

Selective selenol fluorescent probes: design, synthesis, structural determinants, and biological applications.

Selenium (Se) is an essential micronutrient element, and the biological significance of Se is predominantly dependent on its incorporation as selenocysteine (Sec), the genetically encoded 21st amino acid in protein synthesis, into the active site of selenoproteins, which have broad functions, ranging from redox regulation and anti-inflammation to the production of active thyroid hormones. Compared to its counterpart Cys, there are only limited probes for selective recognition of Sec, and such selectivity is strictly restricted at low pH conditions. We reported herein the design, synthesis, and biological evaluations of a series of potential Sec probes based on the mechanism of nucleophilic aromatic substitution. After the initial screening, the structural determinants for selective recognition of Sec were recapitulated. The follow-up studies identified that probe 19 (Sel-green) responds to Sec and other selenols with more than 100-fold increase of emission in neutral aqueous solution (pH 7.4), while there is no significant interference from the biological thiols, amines, or alcohols. Sel-green was successfully applied to quantify the Sec content in the selenoenzyme thioredoxin reductase and image endogenous Sec in live HepG2 cells. With the aid of Sel-green, we further demonstrated that the cytotoxicity of different selenocompounds is correlated to their ability metabolizing to selenols in cells. To the best of our knowledge, Sel-green is the first selenol probe that works under physiological conditions. The elucidation of the structure-activity relationship for selective recognition of selenols paves the way for further design of novel probes to better understand the pivotal role of Sec as well as selenoproteins in vivo.

[Effects of Jiaweisinisan on gastric mucosal ultrastructure and brain-gut axis in a rat model of chronic psychological stress].

OBJECTIVE: To study the effect of Jiaweisinisan (JWSNS), a traditional Chinese herbal medicinal recipe, on gastric mucosal ultrastructure and brain-gut axis in rat models of chronic psychological stress and elucidate the mechanism of JWSNS for ameliorating stress-induced gastrointestinal dysfunction. METHODS: Sixty rats were randomly assigned into normal control group, model group, 3 JWSNS groups (high, moderate, and small doses), and omeprazole group (n=10). Rat models of chronic psychological stress were established by random stressful stimulations, and following the corresponding interventions, plasma adrenocorticotropic hormone (ACTH) and cortisol (CORT) levels were detected using radioimmunoassay, and the mRNA expressions of gastrin receptor in the gastric tissue (GASR) and vasoactive intestinal peptide II receptor (VIPR2) in the jejunal tissue were examined using RT-PCR. Transmission electron microscopy was employed to examine the ultrastructural changes in the gastric mucosa tissue cells of the glandular stomach area and alterations in the intercellular junctions. RESULTS: Electron microscopy revealed obvious damages in gastric mucosal epithelial cell organelles and nuclei in the model rats. These damages were ameliorated after treatments with JWSNS and omeprazole. Compared with the model group, the 3 JWSNS groups and omeprazole group all showed significantly lowered plasma ACTH and CORT levels, increased gastrin receptor mRNA expression and decreased jejunal VIPR2 mRNA expression (P<0.05 or 0.01). CONCLUSION: JWSNS can obviously ameliorate the pathologies of the gastric mucosa cells, regulate the state of brain-gut axis, and modulate the gastric gastrin receptor and jejunal VIPR2 mRNA expressions in rats with chronic psychological stress.

[Effects of Jiaweisinisan dispersion on content and gene expression of gastric tissue GASR and jejunal tissue VIPR2 of physical and mental stress model rat].

OBJECTIVE: To observe the effects of Jiaweisini dispersion (JWSNS) on the ultrastructure of gastric mucosa, the content and gene expression of gastric antrum tissue gastrin receptor (GASR) and jejunal tissue vasoactive intestinal peptide receptor 2 (VIPR2) in chronic stress gastric ulcer rats, and to elucidate its mechanism. METHODS: 60 Wistar rats were randomly divided into normal group, model group, JWSNS large, medium, small dose groups, and omeprazole group, 10 rats in each group. Chronic stress method was used to establish the stress ulcer rat model. The every rat in JWSNS small, medium, large dose groups were gavaged with 0.25, 0.5, 1.0 g/ mL Chinese medicine Decoction on 2 mL respectively daily, rats in omeprazole group were gavaged with 0.3 mg/mL omeprazole solution on 2 mL daily, rats in normal group and model group were gavaged 2 mL NS daily. After modeling was end, transmission electron microscopy (TEM) was used to observe gastric mucosa cells and intercellular connections changes of ultrastructure of glandular stomach area and immunohistochemical method and Real time-PCR method were used to detect the protein content and gene expression changes of gastric antrum tissue GASR and jejunal tissue cell VIPR2. RESULTS: TEM observation demonstrated that in the normal group the gastric mucosa epithelial cells connected compact, cell membrane integrity, cell nuclear shape and size was normal; in model group rats the gastric mucosal cells were severely damaged; the rats in the rest treatment groups were better than those in the model group in different degree. After The treatment of JWSNS and omeprazole, the expression of GASR protein and mRNA in gastric antrum tissue were increased when compared with that of model group (P < 0.05), the expression of VIPR2 protein and mRNA in the jejunum tissue were lower than that of the model group (P < 0.05). The expression of GASR, VIPR2 protein and mRNA in the JWSNS large dose group was closed to the normal group with no significant difference (P > 0.05). And compared with omeprazole group and JWSNS small dose group, expression of GASR protein and mRNA in high dose group rats were increased (P < 0.05), and expression of VIPR2 protein and mRNA were decreased (P < 0.05). CONCLUSION: JWSNS can significantly improve microscopic pathologic morphology of the gastric mucosa cell in gastric ulcer of chronic stress rats models, and can through two aspects of inhibiting damage factor and enhancing defense factor to adjust the content and gene expression of gastric tissue GASR and jejunal tissue VIPR2.