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Calycosin Attenuates Endoplasmic Reticulum Stress Injury in Human Renal Glomerular Endothelial Cells by Inhibiting
Yutian Wang1, Xiaolin Bai1, Ruixia Pei1
1Department of Endocrinology, Rheumatology and Immunology, Xi'an Affiliated Hospital of Shaanxi University of Chinese Medicine (Xi'an Hospital of Traditional Chinese Medicine), Xi'an, Shaanxi, China.
Introduction:
Diabetic Nephropathy (DN), a severe complication, involves endoplasmic reticulum stress. Calycosin, an isoflavone derived from Mongolian Milkvetch Root, may mitigate DN by targeting proto-oncogene tyrosine-protein kinase SRC (SRC)/Spleen Tyrosine Kinase (SYK)-dependent Reactive Oxygen Species (ROS)-mediated ER stress.
Methods:
Human Renal Glomerular Endothelial Cells (HRGECs) were exposed to high glucose to mimic DN, and then treated with calycosin or transfected with SRC overexpression plasmids. Quantitative real-time PCR was performed to assess transfection efficiency, and a western blot was conducted to examine the expression of proteins associated with endothelial dysfunction and endoplasmic reticulum stress. Cell viability, cytotoxicity, apoptosis, and ROS levels were assessed via cell counting kit-8, lactate dehydrogenase assay, flow cytometry, and DCFH-DA probes.
Results:
Calycosin barely affected HRGEC viability in normal glucose medium, but apparently reversed high glucose-induced suppression of viability. High glucose increased cytotoxicity, apoptosis, ROS content, and levels of intercellular cell adhesion molecule-1 (ICAM-1), Glucose-Regulated Protein 78 kD (GRP78), Phosphorylated (P)- Protein Kinase RNA-Like Endoplasmic Reticulum Kinase (PERK)/ total (t)-PERK ratio, C/EBP-Homologous Protein (CHOP), SYK, and SRC protein, which were all counteracted by calycosin. Overexpression of SRC attenuated the effects of calycosin on promoting cell viability and repressing apoptosis, ROS, ICAM-1, and endoplasmic reticulum stress-related proteins in high-glucose-treated HRGECs.
Discussion:
These findings are in line with prior research linking SRC/SYK-ROS signaling to endoplasmic reticulum stress in the context of DN, highlighting the therapeutic potential of calycosin, although the lack of in vivo evidence necessitates future investigation using animal models.
Conclusion:
Calycosin attenuates high glucose-induced endoplasmic reticulum stress injury in HRGECs by inhibiting SRC/SYK-dependent ROS.
Insights
Calycosin, derived from Mongolian Milkvetch Root, protects kidney cells from high glucose damage by reducing endoplasmic reticulum stress via SRC/SYK-dependent reactive oxygen species (ROS) pathways. This study highlights calycosin
Area of Science:
- Nephrology
- Endocrinology
- Cell Biology
Background:
- Diabetic nephropathy (DN) is a severe complication linked to endoplasmic reticulum (ER) stress.
- Calycosin, an isoflavone from Mongolian Milkvetch Root, shows potential in mitigating DN.
- The mechanism may involve targeting SRC/Spleen Tyrosine Kinase (SYK)-dependent Reactive Oxygen Species (ROS)-mediated ER stress.
Purpose of the Study:
- To investigate the protective effects of calycosin on high glucose-induced ER stress in Human Renal Glomerular Endothelial Cells (HRGECs).
- To elucidate the role of SRC/SYK-dependent ROS signaling in calycosin's therapeutic action against DN-related cellular injury.
Main Methods:
- HRGECs were exposed to high glucose and treated with calycosin or SRC overexpression plasmids.
- Quantitative real-time PCR and Western blot analyzed gene and protein expression.
- Cell viability, cytotoxicity, apoptosis, and ROS levels were quantified using standard assays.
Main Results:
- Calycosin reversed high glucose-induced suppression of HRGEC viability, cytotoxicity, apoptosis, and ROS production.
- Calycosin counteracted increases in ICAM-1, GRP78, p-PERK/t-PERK, CHOP, SYK, and SRC proteins.
- SRC overexpression attenuated calycosin's protective effects, indicating SRC's crucial role.
Conclusions:
- Calycosin effectively attenuates high glucose-induced ER stress and injury in HRGECs.
- The protective mechanism involves the inhibition of SRC/SYK-dependent ROS signaling.
- Calycosin demonstrates therapeutic potential for DN, warranting further in vivo investigation.