Bilobalide attenuates lipopolysaccharide­induced HepG2 cell injury by inhibiting TLR4­NF­κB signaling via the PI3K/Akt pathway.

Inflammation is involved in the pathological process underlying a number of liver diseases. Bilobalide (BB) is a natural compound from Ginkgo biloba leaves that was recently demonstrated to exert hepatoprotective effects by inhibiting oxidative stress in the liver cancer cell line HepG2. The anti-inflammatory activity of BB has been reported in recent studies. The major objective of the present study was to investigate whether BB could attenuate inflammation-associated cell damage. HepG2 cells were cultured with lipopolysaccharide (LPS) and BB, and cell damage was evaluated by measuring cell viability using MTT assay. The activity of the NF-κB signaling pathway was assessed by measuring the levels of IκBα, NF-κB p65, phosphorylated (p)-IκBα, p-p65, p65 DNA-binding activity and inflammatory cytokines IL-1ß, IL-6 and TNF-α. A toll-like receptor (TLR)4 inhibitor (CLI-095) was used to detect the involvement of TLR4 in cell injury caused by LPS. In addition, the PI3K/Akt inhibitor LY294002 was applied to explore the involvement of the PI3K/Akt axis in mediating the effects of BB. The results demonstrated that LPS induced HepG2 cell injury. LPS also elevated the levels of p-IκBα, p-p65, p65 DNA-binding activity and inflammatory cytokines. However, CLI-095 significantly attenuated the LPS-induced cell damage and inhibited the activation of NF-κB signaling. BB also dose-dependently attenuated the LPS-induced cell damage, activation of NF-κB signaling and TLR4 overexpression. Furthermore, it was observed that LY294002 diminished the cytoprotective effects of BB on cell injury, TLR4 expression and NF-κB activation. These findings indicated that BB could attenuate LPS-induced inflammatory injury to HepG2 cells by regulating TLR4-NF-κB signaling.

Association between serum Galectin-3 and periodontitis in patients with type 2 diabetes mellitus.

OBJECTIVES: This study aims to investigate the correlation between serum Galectin-3 levels and the risk of periodontitis in patients with type 2 diabetes mellitus (T2DM). METHODS: A total of 140 patients with T2DM admitted to the endocrinology department of Weifang People's Hospital, Affiliated to Weifang Medical College from July 2021 to November 2022 were selected and divided into T2DM without periodontitis group (T2DM group, n=67) and T2DM with periodontitis group (T2DMP group, n=73) according to whether they were combined with periodontitis. In the same period, 65 non-periodontitis volunteers with normal blood glucose were selected as healthy control group (NC group). Blood samples of all subjects were collected, and serum Galectin-3 levels and related laboratory indices were detected and compared among the three groups. RESULTS: Serum Galectin-3 levels in the NC, T2DM, and T2DMP groups were 3.81 (3.49, 4.15), 4.82 (4.25, 5.26), and 6.83 (5.19, 7.28) ng/mL, respectively. After adjusting for the influence of baseline data by multiple linear regression, serum Galectin-3 levels in the T2DMP and T2DM groups were significantly higher than those in the NC group (all P<0.05). Multiple linear regression analysis showed that serum Galectin-3 levels were positively correlated with homeostatic model of the insulin resistance index (ß=0.254, 95%CI:0.089-0.419, P=0.003), glycosylated hemoglobin A1c (ß=0.397, 95%CI: 0.049-0.745, P=0.026), and clinical attachment loss (ß=0.298, 95%CI: 0.024-0.572, P=0.033). After adjusting for the effects of covariates, binary logistic regression showed that serum Galectin-3 levels were significantly associated with the risk of periodontitis in patients with T2DM (OR=2.146, 95%CI: 1.260-3.655, P=0.005). Trend test showed that the risk of periodontitis in patients with T2DM increased with increasing serum Galectin-3 levels (Ptrend=0.011). Receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) of serum Galectin-3 in predicting T2DM periodontitis was 0.861 (95%CI: 0.801-0.920, Z=11.806, P<0.001). CONCLUSIONS: Serum Galectin-3 levels were elevated in patients with T2DM and pe-riodontitis and associated with the risk of periodontitis.

Structural characteristics and immune-enhancing activity of fractionated polysaccharides from Athyrium Multidentatum (Doll.) Ching.

Polysaccharides coded as CP were extracted from Athyrium Multidentatum (Doll.) Ching and then fractionated into five fractions (FP-1, FP-2, FP-3, FP-4 and FP-5). A purified polysaccharide designated as FP-3-4 was prepared from FP-3 by Sephadex G-100 column chromatography. Chemical analysis disclosed that CP and these fractions were heteropolysaccharides and mainly composed of glucose, galactose, arabinose, mannose, rhamnose, xylose, fucose, ribose and uronic acid with different molar ratios. They presented different images of SEM. FP-3-4 was highly branched polymers with sixteen types of linkages. The in vitro immunomodulatory results stated that CP and these fractions could promote macrophage proliferation, enhance macrophage phagocytosis and increase the production of NO, TNF-α, IFN-γ, IL-1ß, IL-6, IL-10 and IL-2, indicating remarkable immune enhancement activities. RNA sequencing analysis revealed that CP and FP-3 induced macrophage activation mainly through MAPK and alternative NF-κΒ signaling pathways via CD14/TLR4 and Dectin-2 receptors, which were verified by RT-qPCR and western blot.

Nanosized FeS/ZnS heterojunctions derived using zeolitic imidazolate Framework-8 (ZIF-8) for pH-universal oxygen reduction and High-efficiency Zn-air battery.

Low-cost, stable, and highly active electrocatalysts for oxygen reduction reaction (ORR), especially for pH-universal ORR, are vital for developing numerous renewable energy devices. Herein, a hierarchical N, S-codoped porous carbon-based catalyst (ZFP-800) coupled with abundant FeS/ZnS heterojunctions was facilely prepared via direct pyrolysis of a Ferrocene-crosslinked pyrrole hydrogel composited with zeolitic imidazolate framework-8 (ZIF-8) templates. Compared with the heterojunction-free catalytic activity, the ZFP-800 catalytic activity was significantly higher in pH-universal ranges. Moreover, the ZFP-800 exhibited competitive ORR performance to commercial Pt/C (20%) in various electrolytes, in terms of onset (Eonset), half-wave potentials (E1/2), limiting current density (JL), durability, and methanol immunity. For instance, it exhibited super ORR catalytic activity on Eonset and E1/2, and exceeded that of the benchmark Pt/C in both the alkaline and neutral media. Furthermore, the application of ZFP-800 as a cathode catalyst in a home-made Zn-air battery demonstrated its operation capability in ambient conditions with a competitive performance on the specific energy density (828 mA·h·gZn-1), maximum discharge power density (205.6 mW·cm-2), rate performance, and the long-term stability (188 h at 5 mA·cm-2). This study can facilitate the development of advanced heterojunction-based materials for renewable energy applications.

A mitochondria-targeted near-infrared fluorescent probe for imaging viscosity in living cells and a diabetic mice model.

A mitochondria-targeted near-infrared fluorescent probe NIR-V with 700 nm emission was designed to monitor cell viscosity changes with high selectivity and sensitivity, which was applied to detect the intracellular viscosity and image pancreatic tissue in a diabetic mouse model. Probe NIR-V provides an effective way to diagnose viscosity related diseases.

Chemical characteristics and cytoprotective activities of polysaccharide fractions from Athyrium Multidentatum (Doll.) Ching.

Five polysaccharide fractions (PS-1, PS-2, PS-3, PS-4 and PS-5) were successfully isolated from Athyrium Multidentatum (Doll.) Ching by anion-exchange column chromatography. Their in vitro cytoprotective activities and the underlying mechanisms were explored in this paper. Chemical analysis suggested that the five polysaccharide fractions were heteropolysaccharides with different molecular weights and monosaccharide compositions. Treatment with these polysaccharide fractions could increase cell viabilities, superoxide dismutase/catalase activities, nitric oxide contents, mitochondrial membrane potential levels and Bcl-2/Bax ratios, and reduce cell apoptosis, intracellular reactive oxygen species production and malondialdehyde contents in H2O2-damaged cells. Moreover, these polysaccharide fractions enhanced the mRNA expression levels of PI3K, Akt, FOXO3a, Nrf2 and HO-1 and PS-4 exhibited the most powerful effects on the mRNA expression of these genes. Current findings suggested that the polysaccharide fractions decreased H2O2-induced apoptosis of HUVECs. The activation of PI3K/Akt/FOXO3a and Nrf2/HO-1 signaling pathways might be involved in the protective mechanisms of the active fractions. The polysaccharides might be one of the key bioactive ingredients of Athyrium Multidentatum (Doll.) Ching for the treatment of oxidative damage.

Neurophysiological mechanisms of bradykinin-evoked mucosal chloride secretion in guinea pig small intestine.

AIM: To investigate the mechanism for bradykinin (BK) to stimulate intestinal secretomotor neurons and intestinal chloride secretion. METHODS: Muscle-stripped guinea pig ileal preparations were mounted in Ussing flux chambers for the recording of short-circuit current (Isc). Basal Isc and Isc stimulated by BK when preincubated with the BK receptors antagonist and other chemicals were recorded using the Ussing chamber system. Prostaglandin E2 (PGE2) production in the intestine was determined by enzyme immunologic assay (EIA). RESULTS: Application of BK or B2 receptor (B2R) agonist significantly increased the baseline Isc compared to the control. B2R antagonist, tetrodotoxin and scopolamine (blockade of muscarinic receptors) significantly suppressed the increase in Isc evoked by BK. The BK-evoked Isc was suppressed by cyclooxygenase (COX)-1 or COX-2 specific inhibitor as well as nonselective COX inhibitors. Preincubation of submucosa/mucosa preparations with BK for 10 min significantly increased PGE2 production and this was abolished by the COX-1 and COX-2 inhibitors. The BK-evoked Isc was suppressed by nonselective EP receptors and EP4 receptor antagonists, but selective EP1 receptor antagonist did not have a significant effect on the BK-evoked Isc. Inhibitors of PLC, PKC, calmodulin or CaMKII failed to suppress BK-induced PGE2 production. CONCLUSION: The results suggest that BK stimulates neurogenic chloride secretion in the guinea pig ileum by activating B2R, through COX increasing PGE2 production. The post-receptor transduction cascade includes activation of PLC, PKC, CaMK, IP3 and MAPK.

Inhibition of leucine aminopeptidase 3 suppresses invasion of ovarian cancer cells through down-regulation of fascin and MMP-2/9.

Leucine aminopeptidase 3 (LAP3) is a cell surface aminopeptidase that catalyzes the hydrolysis of leucine residues from the amino termini of protein or peptide substrates. The over-expression of LAP3 correlates with prognosis and malignant development of several human cell carcinomas. However, the molecular mechanism remains unknown. In this study, we used ES-2 ovarian cancer cell line as a model system to explore the role of LAP3 in regulation of cancer cell invasion by employing a natural LAP3 inhibitor bestatin and LAP3 siRNA. Bestatin inhibited tumor cell migration and invasion in a dose-dependent manner. More interestingly, bestatin down-regulated expression of fascin protein and inhibited activity of fascin promoter luciferase reporter. Both proteome profiler array and Western blot assay showed that bestatin up-regulated the phosphorylation of Hsp27. Furthermore, LAP3 siRNA could up-regulate the phosphorylation of Hsp27 and down-regulate the expression of fascin. Meanwhile, LAP3 siRNA could also down-regulate the phosphorylation of Akt and the expression of MMP-2/9. Taken together, LAP3 could affect the expression of fascin and MMP-2/9 and may act as a potential anti-metastasis therapeutic target.

Hypotensive and Angiotensin-Converting Enzyme Inhibitory Activities of Eisenia fetida Extract in Spontaneously Hypertensive Rats.

Objectives. This study aimed to investigate the antihypertensive effects of an Eisenia fetida extract (EFE) and its possible mechanisms in spontaneously hypertensive rats (SHR rats). Methods. Sixteen-week-old SHR rats and Wistar-Kyoto rats (WKY rats) were used in this study. Rats were, respectively, given EFE (EFE group), captopril (captopril group), or phosphate-buffered saline (PBS) (normal control group and SHR group) for 4 weeks. ACE inhibitory activity of EFE in vitro was determined. The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured using a Rat Tail-Cuff Blood Pressure System. Levels of angiotensin II (Ang II), aldosterone (Ald), and 6-keto-prostaglandin F1 alpha (6-keto-PGF1α ) in plasma were determined by radioimmunoassay, and serum nitric oxide (NO) concentration was measured by Griess reagent systems. Results. EFE had marked ACE inhibitory activity in vitro (IC50 = 2.5 mg/mL). After the 4-week drug management, SHR rats in EFE group and in captopril group had lower SBP and DBP, lower levels of Ang II and Ald, and higher levels of 6-keto-PGF1α and NO than the SHR rats in SHR group. Conclusion. These results indicate that EFE has hypotensive effects in SHR rats and its effects might be associated with its ACE inhibitory activity.

Astragalus Polysaccharide Protects Astrocytes from Being Infected by HSV-1 through TLR3/NF-κB Signaling Pathway.

Astragalus polysaccharide (APS) is the most immunoreactive substance in Astragalus. APS can regulate the body's immunity and is widely used in many immune related diseases. However, till now, there is little information about its contribution to the protection of astrocytes infected by virus. Toll-like receptor 3 (TLR3) is a key component of the innate immune system and has the ability to detect virus infection and trigger host defence responses. This study was undertaken to elucidate the protective effect of APS on herpes simplex virus (HSV-1) infected astrocytes and the underlying mechanisms. The results showed that APS protected the astrocytes from HSV-1 induced proliferation inhibition along with increasing expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) markedly. Moreover, APS significantly promoted the expression of Toll-like receptor 3 (TLR3) and the activation of nuclear factor-κB (NF-κB) in astrocytes. In addition, while astrocytes were pretreated with TLR3 antibody before adding HSV-1 and APS, the expression of TLR3, TNF-α, and IL-6 and the activation of NF-κB decreased sharply. These results indicate that APS can protect astrocytes by promoting immunological function provoked by HSV-1 through TLR3/NF-κB pathway.

[Inhibitory effect of Astragalus polysaccharides on apoptosis of pancreatic beta-cells mediated by Fas in diabetes mellitus rats].

OBJECTIVE: To observe the effects of Astragalus polysaccharides (APS) on the BG, insulin and C-peptide in serum, ultrastructure and Fas expression of pancreatic beta-cells in diabetes mellitus (DM) rats. METHODS: Thirty DM rats induced by streptozotocin (STZ) were randomly divided into three groups: DM group, APS 200 mg/kg group, APS 400 mg/kg group, another 10 normal rats were taken as the control group. The drug was given by intraperitoneal for 6 weeks. The level of BG was determined by ONE TOUCH II machine. The levels of insulin and C-peptide in serum were measured by radioimmunoassay. The expression of Fas was observed by immunohistochemistry. Moreover, the TEM was used to observe the ultrastructure of pancreatic beta-cells. RESULTS: (1) DM rats showed significant increase in BG compared with control group (P < 0.05). APS could decrease the level of FG (P < 0.05). (2) DM rats showed significant decrease in insulin and C-peptide in serum compared with control group (P < 0.05). APS has no appearance effects on the levels of them (P > 0.05). (3) The expression of Fas of beta-cells was significantly increased in DM rats, which was significantly inhibited by APS treatment. (4) It showed degenerative changes of pancreatic beta-cells ultramicroscopic structure of the DM rats, while APS treatment could significantly improve the damage. CONCLUSION: APS exerts its therapeutic effects on DM, which maybe related to the significant decreasing of the Fas expression and inhibiting the apoptosis of beta-cells.