Phosphatase and tensin homolog deficiency induces M2 macrophage polarization by promoting glycolytic activity in endometrial stromal cells

Biology of Reproduction, Journal Year: 2025, Volume and Issue: unknown

Published: March 4, 2025

Endometriosis is a common gynecological disorder, whose pathogenesis remains incompletely understood. Macrophages, key type of immune cell, are pivotal in the context endometriosis. This study seeks to explore interactions between endometriotic cells and macrophages. Quantitative real-time PCR (qRT-PCR) Western blot experiments were employed detect phosphatase tensin homolog (PTEN) expression. Glucose consumption, lactate production, extracellular acidification rate, oxygen consumption rate levels used assess cellular glycolytic capacity. The interaction conditioned media from ectopic endometrial stromal (EESCs) macrophages was investigated through co-culture experiments. expression M2 macrophage marker proteins inflammatory factors detected via qRT-PCR, immunofluorescence staining, enzyme-linked immunosorbent assay. Cellular functions evaluated using Cell Counting Kit-8, 5-Ethynyl-2'-deoxyuridine (EdU), wound healing assays. We found that PTEN deficiency promoted activity EESCs. Simultaneously, it significantly macrophages' polarization toward phenotype, demonstrated by increased markers (differentiation 206 (CD206), CD163, (C-C motif) ligand 22 (CCL22)). Further studies revealed PTEN-deficient EESCs level CCL2 promoting activity, which reversed inhibitor. Moreover, overexpressed facilitated macrophages, while 2-deoxy-d-glucose effect. Furthermore, lactate-facilitated proliferation migration abilities induces EESCs, deepens knowledge pathophysiology endometriosis provides novel insights into its treatment.

Language: Английский

Creation of Genetically Modified Adipocytes for Tissue Engineering: Creatine Kinase B Overexpression Leads to Stimulated Glucose Uptake and Mitochondrial Potential Growth, but Lowered Lipid Synthesis

Life, Journal Year: 2025, Volume and Issue: 15(5), P. 753 - 753

Published: May 8, 2025

Background: The global burden of obesity and type 2 diabetes mellitus is a significant contributor to mortality disability in the modern world. In this regard, modification adipocyte metabolism has been identified as promising approach develop new genetic cellular engineering therapeutics. study, we activate expression creatine kinase B (CKB), key enzyme non-canonical futile cycle regulator energy storage, promote catabolic processes mature adipocytes. Methods: protein-coding sequence CKB was amplified by PCR from Mus musculus brain mRNA. Lentiviral transduction used transfer into Adipocyte analyzed radioisotope monitoring labeled [3H]-2-deoxyglucose [14C]-glucose. Confocal microscopy applied estimate lipid droplets morphology (BODIPY493/503 dye), mitochondrial membrane potential (JC-1 thermogenesis (ERthermAC dye). Results: After lentiviral delivery CKB-coding sequence, mRNA level increased 75-fold protein fivefold. overexpression does not cause changes droplet morphology. Despite this, enhanced glucose uptake reduced synthesis under adrenergic stimulation are detected during overexpression. causes an increase with no effect on Conclusions: have shown that adipocytes allows us obtain high uptake, potency ATP synthesis, suppressed lipogenesis. These genetically modified cells may potentially exhibit favorable metabolic context excessive nutrient utilization.

Language: Английский