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Effects and mechanism of Rictor interference in podocyte injury induced by high glucose
Yan Zeng1, Changbin Xiong1, Yinxiang Chen1
1Department of Nephrology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Abstract:
Rapamycin-insensitive companion of mTOR (Rictor) is a critical effector of mTOR protein complex 2 (mTORC2). The aim of the present study was to investigate the effect of Rictor in the mTORC2 signaling pathway in high glucose (HG)-induced diabetic podocyte injury by silencing the expression of Rictor. In the present study, mouse podocytes were treated with glucose (150 mM) and mannitol (200 mM), the Rictor gene was silenced using small interfering RNA (siRNA). Apoptosis was detected by flow cytometry, whereas podocyte cytoskeletal protein expression was detected by western blotting (WB) and immunofluorescence staining. The results demonstrated that, compared with that in the control group, the podocyte apoptotic rate was significantly increased in the mannitol group (negative group) and the groups that were treated with glucose (model groups). The podocyte apoptotic rate in the model + Rictor siRNA group was significantly decreased compared with that in the negative, model and the model glucose + siRNA negative control (NC) groups. WB indicated that the protein expression levels of podocalyxin and synaptopodin were reduced in the model and model + siRNA NC groups compared with those in the normal control and negative groups. Additionally, the protein expression levels of α-smooth muscle actin (α-SMA) and P-AKT/AKT were increased in the model and model + siRNA NC groups compared with the those in control and negative groups. Compared with those the model and model + siRNA NC groups, the protein expression levels of podocalyxin and synaptopodin were increased, whilst those of the α-SMA and P-AKT/AKT proteins were decreased, in the model + Rictor siRNA group. Results from immunofluorescence analysis were basically consistent with those of WB. Therefore, results of the present study suggest that silencing of the Rictor gene may reduce the damage to podocytes induced by HG, such that the Rictor/mTORC2 signaling pathway may be involved in the remodeling of podocyte actin cytoskeletal in diabetes.
Insights
Silencing Rictor (Rapamycin-insensitive companion of mTOR) reduces high glucose-induced diabetic podocyte injury by modulating the Rictor/mTORC2 pathway and actin cytoskeleton remodeling.
Area of Science:
- Cell Biology
- Nephrology
- Molecular Biology
Background:
- Diabetic nephropathy is a leading cause of kidney failure.
- Podocyte injury is a key event in diabetic kidney disease progression.
- The Rictor-mTORC2 pathway's role in diabetic podocyte injury is not fully understood.
Purpose of the Study:
- To investigate the effect of Rictor silencing on high glucose-induced diabetic podocyte injury.
- To explore the involvement of the Rictor/mTORC2 signaling pathway in podocyte actin cytoskeleton remodeling.
Main Methods:
- Mouse podocytes were treated with high glucose (HG) or mannitol.
- Rictor expression was silenced using small interfering RNA (siRNA).
- Apoptosis was assessed by flow cytometry; cytoskeletal protein expression by Western blotting and immunofluorescence.
Main Results:
- HG significantly increased podocyte apoptosis and α-smooth muscle actin (α-SMA) expression, while decreasing podocalyxin and synaptopodin.
- Silencing Rictor significantly reduced HG-induced podocyte apoptosis and α-SMA levels.
- Rictor silencing increased podocalyxin and synaptopodin expression and reduced P-AKT/AKT levels in HG-treated podocytes.
Conclusions:
- Silencing Rictor protects against high glucose-induced podocyte injury.
- The Rictor/mTORC2 signaling pathway is implicated in diabetic podocyte damage and actin cytoskeleton remodeling.
- Targeting Rictor may offer a therapeutic strategy for diabetic kidney disease.
