Knockdown of TRIM8 Protects HK-2 Cells Against Hypoxia/Reoxygenation-Induced Injury by Inhibiting Oxidative Stress-Mediated Apoptosis and Pyroptosis via PI3K/Akt Signal Pathway.

BACKGROUND: Acute kidney injury (AKI) emerges as an acute and critical disease. Tripartite motif 8 (TRIM8), one number of the TRIM protein family, is proved to participate in ischemia/reperfusion (I/R) injury. However, whether TRIM8 is involved in renal I/R injury and the associated mechanisms are currently unclear. PURPOSE: This study aimed to investigate the precise role of TRIM8 and relevant mechanisms in renal I/R injury. MATERIALS AND METHODS: In this study, human renal proximal tubular epithelial cells (HK-2 cells) underwent 12 hours of hypoxia and 2 h, 3 h or 4 h of reoxygenation to establish an in vitro hypoxia/reoxygenation (H/R) model. The siRNAs specific to TRIM8 (si-TRIM8) were transfected into HK-2 cells to knockdown TRIM8. The cell H/R model included various groups including Control, H/R, H/R+DMSO, H/R+NAC, si-NC+H/R, si-TRIM8+H/R and si-TRIM8+LY294002+H/R. The cell viability and levels of reactive oxygen species (ROS), hydrogen peroxide (H2O2), mRNA, apoptotic proteins, pyroptosis-related proteins and PI3K/AKT pathway-associated proteins were assessed. RESULTS: In vitro, realtime-quantitative PCR and western-blot analysis showed that the mRNA and protein expression of TRIM8 were obviously upregulated after H/R treatment in HK-2 cells. Compared with the H/R model group, knockdown of TRIM8 significantly increased cell viability and reduced the levels of ROS, H2O2, apoptotic proteins (Cleaved caspasebase-3 and BAX) and pyroptosis-related proteins (NLRP3, ASC, Caspase-1, Caspase-11, IL-1ß and GSDMD-N). Western-blot analysis also authenticated that PI3K/AKT pathway was activated after TRIM8 inhibition. The application of 5 mM N-acetyl-cysteine, one highly efficient ROS inhibitor, significantly suppressed the expression of apoptotic proteins and pyroptosis-related proteins. Moreover, the combined treatment of TRIM8 knockdown and LY294002 reversed the effects of inhibiting oxidative stress. CONCLUSION: Knockdown of TRIM8 can alleviate H/R-induced oxidative stress by triggering the PI3K/AKT pathway, thus attenuating pyropyosis and apoptosis in vitro.

Butane-2,3-dione bis-[(4-bromo-benzyl-idene)hydrazone].

The title compound, C(18)H(16)Br(2)N(4), is a linear double Schiff base compound having two parallel 4-bromo-phenyl groups connected across a crystallographic inversion centre by flexible C-C and C=N-N=C bonds and stabilized in the solid state by weak inter-molecular Br⋯Br inter-actions [3.7992 (11) Å], generating an infinite two-dimensional network structure.

Protein recognition onto silica particles using chitosan as intermedium substrate.

A novel molecular imprinting method was used to prepare twice-coated silica particles with specific recognition sites for hemoglobin. Chitosan was used as an intermedium to be coated on silica particles via phase inversion process, and the abundance of exposed amine groups (NH2) were active sites for introducing aldehyde groups. After hemoglobin was covalently immobilized by forming imine bonds with the aldehyde groups, acrylamide was then polymerized onto chitosan-coated silica particles to form the recognition sites. The obtained hemoglobin imprinted [molecularly imprinted polymer (MIP)] beads were characterized by FTIR spectroscopy, thermogravimetric analysis (TG), and scanning electron microscopy (SEM). The MIP particles exhibited selectively adsorption for the imprinted protein compared to the nonselectively adsorption for most of proteins of the nonimprinted (NIP) beads.

A novel solid phase for selective separation of flavonoid compounds.

A novel straightforward approach to selective separation for flavonoid compounds was reported. The solid phase material was prepared by copolymerization using allyl-bromide-modified chitosan as macromonomer, and ethylene glycol dimethacrylate as cross-linker. The material was evaluated by chromatographic analysis; it exhibited high selectivity separation for quercetin and its structural analogues using different mobile phases. The material could directly trap a specific class of compounds including quercetin and kaempferol from the hydrolyzate of Ginkgo biloba extract. These results demonstrated the possibility of direct extraction of certain constituents from herb using this material.

Well-defined lactose-containing polymer grafted onto silica particles.

Reversible addition-fragmentation chain transfer (RAFT) polymerization of 2-O-meth-acryloyloxyethoxyl-(2,3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl)-(1-4)-2,3,6-tri-O-acetyl-beta-D-glucopyranoside (MAEL) was performed directly in CHCl3 solutions using cumyl dithiobenzoate (CDB) as the chain transfer agent to give well-defined glycopolymers. The chemical composition and structure of the glycopolymer were characterized by 1HNMR, FTIR, and SEC. The living glycopolymer chains were subsequently grafted onto gamma-methacryloxypropyl-trimethoxy (MPTMS) modified silica particles. The acetyl groups of the poly(MAEL) grafted onto the silica gel particles were converted to the hydroxyl groups with CH3ONa/CH3OH, thus obtaining silica gel particles modified with well-defined lactose-carrying polymer.

Hemoglobin recognition by imprinting in semi-interpenetrating polymer network hydrogel based on polyacrylamide and chitosan.

Semi-interpenetrating polymer network (semi-IPN) hydrogel was prepared to recognize hemoglobin, by molecularly imprinted method, in the mild aqueous media of chitosan and acrylamide in the presence of N,N'-methylenebisacrylamide as the cross-linking agent. The hydrogel obtained has been investigated by using thermal analysis, X-ray diffraction, differential scanning calorimetry (DSC), and environmental scanning electron microscope (ESEM). Langmuir analysis showed that an equal class of adsorption was formed in the hydrogel, and the adsorption equilibrium constant and the maximum adsorption capacity were evaluated to be 4.27 g/mL and 36.53 mg/g wet hydrogel, respectively. The imprinted semi-IPN hydrogel has a much higher adsorption capacity for hemoglobin than the nonimprinted hydrogel with the same chemical composition and also has a higher selectivity for the imprinted molecule.

Chitosan beads as molecularly imprinted polymer matrix for selective separation of proteins.

Two kinds of molecularly imprinted polymers were prepared using hemoglobin as the imprinting molecule, acrylamide as the functional monomer, chitosan beads and maleic anhydride-modified chitosan beads as matrixes, respectively. Static adsorbing experimental results showed that an equal class of adsorption was formed in the imprinted polymers and the adsorption equilibrium constant and the maximum adsorption capacity were evaluated. Chromatographic characteristics showed that the column bedded with the hemoglobin imprinted beads could separate hemoglobin and bovine serum albumin effectively from their mixture, which indicates that the imprinted beads have very higher selectivity for hemoglobin than the non-imprinted with the same chemical composition.

[Dynamic double modulation with step scan FTIR spectroscopy on polyurethane film].

The principle of step scan FTIR spectroscopy and the relationship between phase modulation and sample modulation were described in detail. Demodulation of the frequency domain detector signal was discussed and illustrated using double digital signal processing (DSP2) algorithm. In-phase and quadrature dynamic spectra of polyurethane with 5 Hz stretching under 100 Hz phase modulation as compared to the normal spectrum were presented. The spectral resolution of the overlapped features had greatly been enhanced.