Related Experiment Video
Updated: Aug 22, 2026
Optimizing Isolation and Purification of Murine Glomerular Mesangial Cells
Published on: March 7, 2025
Glycated albumin induces superoxide generation in mesangial cells
Chong Woo Yoo1, Chi Young Song, Bong Cho Kim
1Department of Pathology, Seoul National University College of Medicine, Seoul, Korea.
Background/Aims:
Reactive oxygen species are involved in the pathogenesis of diabetic nephropathy. Amadori-modified glycated albumin modulates signaling pathways in mesangial cells that contribute to the development of diabetic nephropathy. However, the effects of glycated albumin on mesangial cell superoxide (O2-) production are unknown. Thus, we examined whether glycated albumin induces mesangial cell O2- generation and whether increased O2- production elicits cell growth.
Methods:
Quiescent human mesangial cells (HMC) were exposed to bovine serum albumin (BSA) or glycated BSA (Gly-BSA) with or without diphenylene iodonium (DPI) or apocynin, inhibitors of NAD(P)H oxidase, GF109203X (GFX), a protein kinase C (PKC) inhibitor.
Results:
Gly-BSA increased PKC activity, particularly PKC-alpha and -alpha1, within 15 min of incubation with HMC, which decreased to the control value at 2 h. Gly-BSA incubated with HMC increased O2- production by 2 times vis-á-vis BSA-treated cells. The Gly-BSA-induced increased O2- generation was suppressed by DPI or GFX. Gly-BSA significantly increased mesangial [3H]-leucine incorporation, whereas these processes were abrogated by DPI, apocynin or GFX.
Conclusions:
Gly-BSA induces PKC/NAD(P)H oxidase-dependent O2- production in HMC, which in turn results in cell hypertrophy. Thus, O2- induced by glycated albumin might cause mesangial cell alterations in diabetes participating in the pathophysiology of diabetic nephropathy.
Insights
Glycated albumin (Gly-BSA) triggers reactive oxygen species production in kidney cells via protein kinase C (PKC) and NAD(P)H oxidase, leading to cell growth implicated in diabetic nephropathy.
Area of Science:
- Nephrology
- Cell Biology
- Biochemistry
Background:
- Reactive oxygen species (ROS) are implicated in diabetic nephropathy pathogenesis.
- Amadori-modified glycated albumin influences mesangial cell signaling pathways relevant to diabetic nephropathy.
- The specific impact of glycated albumin on mesangial cell superoxide (O2-) production was previously unknown.
Purpose of the Study:
- To investigate if glycated albumin induces superoxide (O2-) generation in human mesangial cells (HMC).
- To determine if elevated O2- production stimulates HMC growth.
- To elucidate the signaling pathways involved in glycated albumin-induced O2- production.
Main Methods:
- Human mesangial cells (HMC) were treated with bovine serum albumin (BSA) or glycated BSA (Gly-BSA).
- Inhibitors of NAD(P)H oxidase (diphenylene iodonium [DPI], apocynin) and protein kinase C (PKC) inhibitor (GF109203X [GFX]) were used.
- Superoxide (O2-) production and [3H]-leucine incorporation (a measure of protein synthesis/cell growth) were assessed.
Main Results:
- Gly-BSA rapidly increased PKC activity in HMC.
- Gly-BSA treatment resulted in a twofold increase in O2- production compared to BSA.
- Inhibition of PKC or NAD(P)H oxidase abrogated Gly-BSA-induced O2- generation and cell growth.
Conclusions:
- Gly-BSA stimulates O2- production in HMC through a pathway involving PKC and NAD(P)H oxidase.
- This increased O2- production leads to mesangial cell hypertrophy.
- O2- generated by glycated albumin may contribute to mesangial cell dysfunction in diabetic nephropathy.
