Abstract
Purpose :
To identify the key proteins involved in the pathological mechanism of diabetic cataract (DC) through proteomic analysis, regulate the signaling pathways, and discover potential targets for preventing DC.
Methods :
Anterior capsule(AC) from DC and age-related cataract(ARC) patients were collected for 4D-Data independent acquisition proteomic analysis to identified differentially expressed proteins (DEPs). Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used for the analysis, and parallel reaction monitoring was used to validate the proteins.The transcriptome and protein levels of SH2B1 were validated in human AC, lens of diet-induced obesity mouse model (DIO) and high glucose (HG) cultured HLECs. CCK8 assay and wound healing assay were used to detect cell proliferation and migration, flow cytometry was used to detect mitochondrial membrane potential, and TUNAL staining was used to observe apoptosis. The protein expression levels of Bcl2 and Caspase3 were detected by elisa kits. The expression levels of P38MAPK signaling pathway proteins were detected. Overexpression and knockdown of proteins in cells with lentiviruses to confirm their roles in the signaling pathways.
Results :
SH2B1 may be a key proteinin DC and its transcription and protein expression levels were down-regulated in various samples.Under HG conditions, cell proliferation and migration , mitochondrial membrane potential decreased, and Bcl2 decreased, while Caspase3 increased. Overexpression of SH2B1 promoted cell proliferation and migration, reversed the decrease of mitochondrial membrane potential, and the increase of Bcl2 and Caspase3. The P38MAPK signaling pathway was inhibited in HG. The regulation of SH2B1 can activate or inhibit the P38MAPK signaling pathway. The phosphorylation of CREB at ser133 was significantly inhibited in the DC anterior capsule and LECs exposed to HG, which activated the mitochondrial apoptotic pathway.
Conclusions :
High glucose can lead to increased mitochondrial apoptosis in lens epithelial cells, and the SH2B1 p38MAPK/CREB activation signal channel plays an important role in this process.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.