Oligodendroglia-to-pericyte conversion after lipopolysaccharide exposure is gender-dependent.

To investigate the sex-dependent differentiation of Sox10 cells and their response to pathological conditions such as lipopolysaccharide (LPS) exposure or ischemia, we utilized Sox10 Cre-ERT2, tdTomato mice. Tamoxifen administration induced the expression of red fluorescent protein (RFP) in these cells, facilitating their subsequent tracking and analysis after LPS injection and ischemia via immunofluorescence staining. Propidium iodide (PI) was injected to label necrotic cells following LPS administration. We found that the conversion of Sox10 cells to pericytes in female mice was significantly higher than in male mice, especially in those exposed to LPS. After LPS injection, the number of PI+ necrotic cells were significantly greater in females than in males. Moreover, RFP+ cells did not co-localize with glial fibrillary acidic protein (GFAP) or cluster of differentiation 11b (CD11b). Similarly, after brain ischemia, RFP+ cells did not express cluster of differentiation 13 (CD13), neuronal nuclei (NeuN), GFAP, or ionised calcium binding adaptor molecule 1 (Iba-1). These findings indicate that the conversion of Sox10 cells to pericytes following LPS exposure is sex-dependent, with neither male nor female groups showing differentiation into other cell types after LPS exposure or under ischemic conditions. The differences in LPS-induced necrosis of pericytes between sexes may explain the variations in the conversion of Sox10 cells to pericytes in both sexes.

Palmitoleic acid protects microglia from palmitate-induced neurotoxicity in vitro.

Although palmitoleic acid (POA) is a lipokine with beneficial effects on obesity and is produced as a byproduct from the manufacture of prescription omega-3 fatty acids, its role in nervous system inflammation is still unknown. This study aims to examine the mechanisms and protective effects of POA against palmitic acid (PA)-induced microglial death. PA-induced microglial death was used as a model for POA intervention. Various inhibitors were employed to suppress potential routes of PA entry into the cell. Immunofluorescence staining and Western blotting were conducted to elucidate the protective pathways involved. The results suggest POA has the potential to eliminate PA-induced lactate dehydrogenase (LDH) release, which decreases the overall number of propidium iodide (PI)-positive cells compared with control. Moreover, POA has the potential to significantly increase lipid droplets (LDs) in the cytoplasm, without causing any lysosomal damage. POA inhibited both canonical and non-canonical gasdermin D (GSDMD)-mediated pyroptosis and gasdermin E (GSDME)-mediated pyroptosis, which PA typically induces. Additionally, POA inhibited the endoplasmic reticulum (ER) stress and apoptosis-related proteins induced by PA. Based on the findings, POA can exert a protective effect on microglial death induced by PA via pathways related to pyroptosis, apoptosis, ER stress, and LDs.

Palmitate lipotoxicity is closely associated with the fatty acid-albumin complexes in BV-2 microglia.

Palmitic acid (PA) is considered a major contributor to the inflammation in many metabolic diseases; however, this role has been questioned recently for the complicated procedures in preparing PA-bovine serum albumin (BSA) complex. This study is aimed to evaluate the effect of PA-BSA complexing methods on cell viability and inflammatory responses of BV-2 cells. Three commercially available BSA brands and two types of solvents were compared for their effects on the expression of inflammatory cytokines. Three commonly used proportions of PA-BSA were tested for cell viability and inflammatory responses. We found that all the three types of BSA were proinflammatory. Both ethanol and isopropanol dampened inflammation except that 1% isopropanol treatment increased the IL-1ß level by 26%. When reducing the BSA content in PA-BSA solutions from 3:1 to 5:1, a marked increase in cell viability (11%) was seen. To our surprise, reducing BSA content in PA-BSA solutions from 5:1 to 10:1 decreased cell viability by 11%. The 5:1 group exhibited the lowest inflammatory profile. Either PA-BSA or BSA alone increased the entry of LPS to the cytosol, which further caused pyroptosis. In summary, we found 5:1 (PA:BSA) to be the best binding ratio for studying inflammation in BV-2 microglia. The presence of LPS in the cytosol in the context of BSA might be the reason for confounding results from palmitate studies.

A single dose of lipopolysaccharide elicits autofluorescence in the mouse brain.

One hallmark of aging is autofluorescence (AF) in the brain. However, the underlying mechanism for inducing AF remains unknown. This study aims to determine the cause(s) of this phenomenon. The endogenous expression pattern of AF in mice was examined at differing ages. Intraperitoneal injection of a single dose of lipopolysaccharide (LPS) was performed to induce AF. Copper sulfate was applied to remove AF to allow for further immunofluorescence staining. AF appeared in the mouse brain as early as 3 months of age. In the cortex, AF occurs in the lysosomes of microglia, astrocytes, endothelial cells, and oligodendrocyte lineage cells and its prevalence increases with age. Interestingly, AF never occurs in the pericytes of young or aged brains. LPS administration resulted in a rapid and marked induction of brain AF, similar to the normal aging process. Finally, age-related and induced AF can be eliminated by low concentrations of copper sulfate solution. This pre-treatment is safe for aging and lineage tracing studies. These findings depict that AF in the brain could be associated with the innate immune response against Gram-negative bacteria infection.

Understanding the impact of pectin physicochemical variation on browning of simulated Maillard reaction system in thermal and storage processing.

Maillard reaction browning is one of the quality deterioration in dried fruit products, but how pectin affects Maillard reaction in the fruit drying and storage process is not clear. This study aimed at investigating the mechanism of pectin variation impact on the browning of Maillard reaction by using simulated system (l-lysine, d-fructose and pectin) in thermal (60 °C and 90 °C for 8 h) and storage (37 °C for 14 days) process. Results showed that apple pectin (AP) and sugar beet pectin (SP) significantly enhanced the browning index (BI) of the Maillard reaction system by 0.01 to 134.51 in the thermal and storage processes, respectively, which were methylation degree of pectin-dependent. The pectin depolymerization product participated Maillard reaction by reacting with l-lysine, and increasing the 5-hydroxymethyl furfural (5-HMF) content (1.25-11.41-fold) and Abs420nm (0.01-0.09). It also produced a new product (m/z 225.1245), which finally increased browning level of the system.

Understanding the impact of pectin on browning of polyphenol oxidation system in thermal and storage processing.

Browning of some processed fruit products was affected not only by polyphenol oxidation but also by cell wall polysaccharides (pectin). The study was performed to understand the mechanism of browning in the pectin system. The catechin/chlorogenic acid oxidation system in three pectins significantly enhanced their browning during thermal storage with pectin structure- and concentration-dependent. Particularly, the structural and physicochemical properties of pectin were examined to determine its effects on the kinetics of polyphenol oxidation and the stability of free polyphenols. Moreover, pectin impacted the fluorescence characteristics of polyphenols by cross-linking with the aromatic ring of polyphenols. In turn, the interaction between polyphenols and pectin impacted the chemical bond vibration of pectin, thereby affecting its optical features and browning. The correlation analysis revealed that the monosaccharide composition, Ratio 1, Ratio 2, Ratio 3, methyl esterification, ζ-potential, and polydispersity index of pectin were significantly correlated with the browning of the pectin-polyphenol oxidation system.