Agar-gelatin Maillard conjugates used for Pickering emulsion stabilization.

A few protein- and polysaccharide-based particles have shown promising potential as stabilizers in multi-phase food systems. By incorporating polymer-based particles and modifying the wettability of colloidal systems, it is possible to create particle-stabilized emulsions with excellent stability. A Pickering emulsifier (AGMs) with better emulsifying properties was obtained by the Maillard reaction between acid-hydrolysed agar and gelatin. Laser confocal microscopy imaging revealed that AGMs particles can be used as solid emulsifiers to produce a typical O/W Pickering emulsion, with AGMs adsorbing onto the droplet surface to form a dense interfacial layer. Cryo-scanning electron microscopy analysis showed that AGMs self-assembled into a three-dimensional network structure, which prevented droplets aggregation through strong spatial site resistance, contributing to emulsion stabilization. These emulsions exhibited stability within a pH range of 1 to 11, NaCl concentrations not exceeding 300 mM, and at temperatures below 80 °C. The most stable emulsion oil-water ratio was 6:4 at a particle concentration of 0.75 % (w/v). AGMs-stabilized Pickering emulsion was utilized to create a semi-solid mayonnaise as a replacement for hydrogenated oil. Rheological analysis demonstrated that low-fat mayonnaise stabilized with AGMs exhibited similar rheological behavior to traditional mayonnaise, offering new avenues for the application of Pickering emulsions in the food industry.

Study on the function of Huazhuo Jiedu Decoction in promoting the homing of bone marrow mesenchymal stem cells and contributing to the treatment of ulcerative colitis.

Objective: To study the function of Huazhuo Jiedu Decoction (HZJD) in promoting the homing of bone marrow mesenchymal stem cells (BMSCs) and contributing to the reconstruction of the intestinal mucosal barrier in ulcerative colitis. Methods: Bone mesenchymal stem cells derived from mice were isolated and cultured, osteogenic and adipogenic assays to study the differentiation ability of BMSCs, and flow cytometry was used to detect the surface marker of the third generation cells. 30 mice were selected and divided into blank group, model group, HZJD group, BMSCs group, and HZJD combined with BMSCs group. Mouse colon length, body weight, and DAI score were used to assess efficacy. The levels of IL-6, IL-1ß, TNF-α, and IFN-γ in serum were measured by ELISA. BMSCs transfected with GFP were used to mark the homing of BMSCs in mice. The BMSCs tagging protein CD90+/CD29+ was detected by immunofluorescence. H&E staining detects damage to the colon and the inflammatory response. The expression levels of claudin-2, claudin-4, occludin, and ZO-1 in colon tissues were detected by Western blot. Results: After subculture, the cell grew with adherence. Flow cytometry showed that the cells were CD73+/CD90+/CD29+/CD45-/CD34-, which belonged to bone mesenchymal stem cells. ELISA showed that the treatment with HZJD and BMSCs suppressed the DSS-induced inflammatory response. BMSCs carrying GFP can be detected in intestinal tissues. Immunofluorescence showed that the HZJD combined with the BMSCs group had more BMSCs homing to the colonic tissue. The results of H&E and Western blot showed that DSS-induced intestinal mucosal damage in UC mice was repaired by HZJD and BMSCs, and the abnormal tight junction proteins claudin-2, claudin-4, occludin, and ZO-1 were normalized. Conclusion: HZJD has a therapeutic effect on ulcerative colitis by promoting the migration of BMSCs to ulcers of the colon and contributing to the reconstruction of the intestinal mucosal barrier in ulcerative colitis.

Diosmetin alleviated cerebral ischemia/reperfusion injury in vivo and in vitro by inhibiting oxidative stress via the SIRT1/Nrf2 signaling pathway.

Cerebral ischemia/reperfusion (I/R) injury is caused by blood flow recovery after an ischemic stroke, and effective treatments targeting I/R injury are still insufficient. Oxidative stress is known to play a pivotal role in the pathogenesis of cerebral I/R injury. Previous studies have revealed that diosmetin could protect against oxidative stress in cerebral I/R injury, but the underlying mechanisms have not been fully revealed. The present study was undertaken to investigate the effects and mechanisms of action of diosmetin on cerebral I/R injury. In vivo, rats were orally gavaged with diosmetin for seven days, and middle cerebral artery occlusion (MCAO) was established to simulate cerebral I/R injury. The neurological deficit score, cerebral infarct volume, and cortical pathological lesions were measured. In vitro, PC12 cells were exposed to oxygen-glucose deprivation/reoxygenation (OGD/R). To clarify the mechanism, the SIRT1 inhibitor EX527 and the small interfering RNA (siRNA) of SIRT1 were used to downregulate the SIRT1 protein level, respectively. The contents of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and malondialdehyde (MDA) were determined with commercial kits. The protein expressions of SIRT1, total Nrf2 (T-Nrf2), nucleus Nrf2 (N-Nrf2), NQO1 and HO-1 were measured by western blotting. The results showed that diosmetin pretreatment improved neurological outcomes, decreased the cerebral infarct volume and pathological lesions, and inhibited oxidative stress in cerebral I/R rats. In PC12 cells, diosmetin increased cell viability, reduced lactate dehydrogenase (LDH) release and reactive oxygen species (ROS) level, and inhibited oxidative stress. Besides, diosmetin increased the protein expressions of SIRT1, T-Nrf2, N-Nrf2, NQO1 and HO-1 both in vivo and in vitro. However, administration of EX527 or silencing the SIRT1 gene with its siRNA eliminated the beneficial effects of diosmetin. Meanwhile, inhibition of SIRT1 decreased the levels of Nrf2 and the protein expressions of its downstream antioxidants NQO1 and HO-1. In conclusion, our data suggested that diosmetin could attenuate cerebral I/R injury by inhibiting oxidative stress via the SIRT1/Nrf2 signaling pathway.

In Silico and In Vivo Studies on the Mechanisms of Chinese Medicine Formula (Gegen Qinlian Decoction) in the Treatment of Ulcerative Colitis.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease, and Gegen Qinlian Decoction (GQD), a Chinese botanical formula, has exhibited beneficial efficacy against UC. However, the mechanisms underlying the effect of GQD still remain to be elucidated. In this study, network pharmacology approach and molecular docking in silico were applied to uncover the potential multicomponent synergetic effect and molecular mechanisms. The targets of ingredients in GQD were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Bioinformatics Analysis Tool for Molecular mechANism of TCM (BATMAN-TCM) database, while the UC targets were retrieved from Genecards, therapeutic target database (TTD) and Online Mendelian Inheritance in Man (OMIM) database. The topological parameters of Protein-Protein Interaction (PPI) data were used to screen the hub targets in the network. The possible mechanisms were investigated with gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Molecular docking was used to verify the binding affinity between the active compounds and hub targets. Network pharmacology analysis successfully identified 77 candidate compounds and 56 potential targets. The targets were further mapped to 20 related pathways to construct a compound-target-pathway network and an integrated network of GQD treating UC. Among these pathways, PI3K-AKT, HIF-1, VEGF, Ras, and TNF signaling pathways may exert important effects in the treatment of UC via inflammation suppression and anti-carcinogenesis. In the animal experiment, treatment with GQD and sulfasalazine (SASP) both ameliorated inflammation in UC. The proinflammatory cytokines (TNF-α, IL-1ß, and IL-6) induced by UC were significantly decreased by GQD and SASP. Moreover, the protein expression of EGFR, PI3K, and phosphorylation of AKT were reduced after GQD and SASP treatment, and there was no significance between the GQD group and SASP group. Our study systematically dissected the molecular mechanisms of GQD on the treatment of UC using network pharmacology, as well as uncovered the therapeutic effects of GQD against UC through ameliorating inflammation via downregulating EGFR/PI3K/AKT signaling pathway and the pro-inflammatory cytokines such as TNF-α, IL-1ß and IL-6.

Electroacupuncture ameliorates cerebral ischemia/reperfusion injury by suppressing autophagy via the SIRT1-FOXO1 signaling pathway.

Cerebral ischemia/reperfusion (CIR) injury occurs when blood flow is restored in the brain, causing secondary damage to the ischemic tissues. Previous studies have shown that electroacupuncture (EA) treatment contributes to brain protection against CIR injury through modulating autophagy. Studies indicated that SIRT1-FOXO1 plays a crucial role in regulating autophagy. Here we investigated the mechanisms underlying the neuroprotective effect of EA and its role in modulating autophagy via the SIRT1-FOXO1 signaling pathway in rats with CIR injury. EA pretreatment at "Baihui", "Quchi" and "Zusanli" acupoints (2/15Hz, 1mA, 30 min/day) was performed for 5 days before the rats were subjected to middle cerebral artery occlusion, and the results indicated that EA pretreatment substantially reduced the Longa score and infarct volume, increased the dendritic spine density and lessened autophagosomes in the peri-ischemic cortex of rats. Additionally, EA pretreatment also reduced the ratio of LC3-II/LC3-I, the levels of Ac-FOXO1 and Atg7, and the interaction of Ac-FOXO1 and Atg7, but increased the levels of p62, SIRT1, and FOXO1. The above effects were abrogated by the SIRT1 inhibitor EX527. Thus, we presume that EA pretreatment elicits a neuroprotective effect against CIR injury, potentially by suppressing autophagy via activating the SIRT1-FOXO1 signaling pathway.

Protection of the Geum japonicum Thunb. var. chinense extracts against oxygen-glucose deprivation and re-oxygenation induced astrocytes injury via BDNF/PI3K/Akt/CREB pathway.

Geum japonicum Tunb. var. chinense (GJ) is a traditional Chinese medicine usually used for the alleviation of dizziness and headache. Previous studies have reported that the GJ extracts could alleviate cerebral I/R injury by reducing apoptosis in vivo. To further elucidate the positive role and underlying mechanism of the GJ extracts in cerebral I/R injury, the current study investigated the effects of the GJ extracts on oxygen-glucose deprivation and re-oxygenation (OGD/R)-induced astrocytes injury in light of BDNF/PI3K/Akt/CREB signaling pathway with seropharmacological method. In the present study, the LC-MS profiling of the GJ extracts, obtain by reflux extraction, led to the identification of three possible active components were 5-desgalloylstachyurin, tellimagrandin II (TG II) and 3,4,5-Trihydroxybenzaldehyde (THBA). Drug-containing serum was collected from rats given different doses of the GJ extracts (0, 1.75 g/kg, 7 g/kg). Data indicated that the GJ extracts could increase the cell viability and decrease apoptosis and the expression of glial fibrillary acidic protein (GFAP) in OGD/R-induced astrocytes. In addition, the detection of apoptosis-related factors showed that the GJ extracts could obviously increase the expression of Bcl-2 and reduce the expression of Bax, Caspase-3 and cleaved-Caspase-3. Furthermore, the GJ extracts markedly increased the expression of BDNF, TrkB, PI3K, p-Akt and p-CREB. All these effects of the GJ extracts could be significantly reversed by LY294002, an inhibitor of PI3K. These data indicated that the GJ extracts could protect astrocytes against OGD/R-induced injury by inhibiting astrocytes reactivity and apoptosis, owing to the activation of the BDNF/PI3K/Akt/CREB pathway. The results support the application of the GJ extracts in the treatment of ischemic stroke and other ischemic encephalopathy.

[Electroacupuncture pretreatment alleviated cerebral ischemia-reperfusion injury via suppressing autophagy in cerebral cortex tissue in rats].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) preconditioning on autophagy in cerebral cortex tissues of rats with cerebral ischemia-reperfusion injury (CIRI), so as to explore its mechanisms underlying improvement of CIRI. METHODS: Thirty-three male Sprague-Dawley rats were randomly divided into sham operation, model and EA groups (n=11 in each group). EA (2 Hz/15 Hz, 1 mA) was applied to "Baihui"(GV20), "Quchi" (LI11) and "Zusanli" (ST36) for 30 min, once daily for 5 days, followed by establishment of CIRI model by occlusion of the middle cerebral artery (MCAO) for 1.5 h and reperfusion for 24 h. The neurological deficit score was assessed in reference to Longa's methods, and the infarct volume assessed by 2,3,5-triphenyltetrazolium chloride staining. The density of dendrite spines of neurons in the ischemic cerebral cortex tissue was detected by Golgi's staining, the autophagosome observed by electron microscopy, and the expression levels of microtubule-associated protein 1 light chain 3 (LC3) and p62 (a selective autophagy substrate) were detected by Western blot. RESULTS: Compared with the sham operation group, the neurological deficit score and infarct volume were significantly increased (P<0.01), the number of autophagosomes and the ratio of LC3-Ⅱ/LC3-Ⅰ also significantly increased (P<0.01), while the expression level of p62 was notably decreased in the model group (P<0.01). Following the intervention and in comparison with the model group, the neurological deficit score and infarct volume were significantly reduced (P<0.01), the number of autophagosomes and the ratio of LC3-Ⅱ/LC3-Ⅰ obviously decreased (P<0.01), and the expression of p62 was significantly up-regulated in the EA group (P<0.01). CONCLUSION: EA pretreatment is effective in improving CIRI in rats, which may be realized through suppressing autophagy in the ischemic cerebral cortex tissue.

The Potential Role of MicroRNA-124 in Cerebral Ischemia Injury.

Cerebral ischemia injury, the leading cause of morbidity and mortality worldwide, initiates sequential molecular and cellular pathologies that underlie ischemic encephalopathy (IE), such as ischemic stroke, Alzheimer disease (AD), Parkinson's disease (PD), epilepsy, etc. Targeted therapeutic treatments are urgently needed to tackle the pathological processes implicated in these neurological diseases. Recently, accumulating studies demonstrate that microRNA-124 (miR-124), the most abundant miRNA in brain tissue, is aberrant in peripheral blood and brain vascular endothelial cells following cerebral ischemia. Importantly, miR-124 regulates a variety of pathophysiological processes that are involved in the pathogenesis of age-related IE. However, the role of miR-124 has not been systematically illustrated. Paradoxically, miR-124 exerts beneficial effects in the age-related IE via regulating autophagy, neuroinflammation, oxidative stress, neuronal excitability, neurodifferentiation, Aß deposition, and hyperphosphorylation of tau protein, while it may play a dual role via regulating apoptosis and exerts detrimental effects on synaptic plasticity and axonal growth. In the present review, we thus focus on the paradoxical roles of miR-124 in age-related IE, as well as the underlying mechanisms. A great understanding of the effects of miR-124 on the hypoxic-ischemic brain will open new avenues for therapeutic approaches to protect against cerebral ischemia injury.

[Pyroptosis induced by zinc oxide nanoparticles in A549 cells].

OBJECTIVE: To explore whether exposure to zinc oxide nanoparticles (ZnO-NPs) induced pyroptosis in human alveolar type II epithelial cells (A549). METHODS: A549 cells were stimulated with different concentrations of ZnO-NPs (10 and 20 microg/ml) for 8 and 24h, respectively. The activity of lactic dehydrogenase (LDH) and the levels of interleukin-1beta (IL-1beta) in cell culture media were respectively measured by LDH activity kit and human IL-1beta ELISA kit. The activity of caspase-1 in A549 cells was detected by caspase-1 colorimetric assay kit. RESULTS: At 8h exposure to ZnO-NPs, there was no significant difference in the activity of LDH in the cell culture media among the ZnO-NPs-treated and control groups, but the activity of caspase-1 and the levels of IL-1beta in A549 cells were significantly increased in 20 microg/ml ZnO-NPs group compared to that in the control group (P < 0.05). Levels of IL-1beta and activity of LDH in the groups treated with ZnO-NPs (10 or 20 microg/ml) were significantly higher than that in the control group after 24 h exposure to ZnO-NPs, but there was no significant difference in the activity of caspase-1 among ZnO-NPs and control group. CONCLUSION: ZnO-NPs (20 microg/ml) treatment induces early increase in caspase-1 activity followed by the increase in LDH activity and IL-1 levels, indicative of pyroptosis in A549 cells.

[Coal tar pitch smoke extract-induced pyroptosis in human bronchial epithelial cells].

OBJECTIVE: To explore whether coal tar pitch smoke extract (CTP) induced pyroptosis in human bronchial epithelial cells (BEAS-2B). METHODS: BEAS-2B cells were treated with different concentrations of CTP (1, 3 µg/ml) for 8h and 24 h, respectively. Lactic dehydrogenase (LDH) activity and interleukin-1 beta (IL-1ß) levels in the supernatants of cell culture media were measured with LDH activity or human IL-1ß ELISA kit, respectively. The activity of Caspase-1 was measured with Caspase-1 colorimetric assay kit. RESULTS: The activity of caspase-1 in 1 and 3 µg/ml CTP groups were (9.29 ± 0.30) and (8.67 ± 0.59) µmol/ml respectively which were both significantly increased compared to that (7.42 ± 0.59) µmol/ml in the control group (P < 0.05) after 8 h exposure, but there was no significant difference in the activity of LDH and levels of IL-1ß in the cell culture media among the CTP and control groups. 24 h after exposure, the activity of LDH in the CTP (1, 3 µg/ml) groups were (1323.03 ± 28.53) and (1148.45 ± 16.42) U/dl respectively which were significantly higher than that (1091.93 ± 26.64) U/dl in the control group (P < 0.05), and the levels of IL-1ß in the CTP (1 and 3 µg/ml) groups were (125.37 ± 25.00) pg/ml and (92.04 ± 19.09) pg/ml respectively which were significantly higher than that (46.20 ± 14.43) pg/ml in the control group (P < 0.05), but there was no significant difference in the activity of Caspase-1 among CTP and control groups (P < 0.05). CONCLUSION: CTP treatment induced early increase in caspase-1 activity followed by the increase in LDH activity and IL-1 levels, indicative of pyroptosis in human bronchial epithelial cells.