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Mechanism of Kemeng Fang's Inhibition of Podocyte Apoptosis in Rats with Membranous Nephropathy through the PI3K/AKT Signaling Pathway
Published on: August 23, 2024
[Effect and molecular mechanism of interferon-α on podocyte apoptosis induced by hepatitis B virus X protein]
Yong-Hong Sun1, Xiao-Yan Lei, Xing-Xing Chen
1Department of Pediatrics, Gansu Province People's Hospital, Lanzhou 730000, China. leixiaoyan601@163.com.
Objective:
To investigate the effect and molecular mechanism of interferon-α (INF-α) on the apoptosis of the mouse podocyte cell line MPC5 induced by hepatitis B virus X (HBx) protein.
Methods:
MPC5 cells were transfected with the pEX plasmid carrying the HBx gene. RT-PCR was used to measure the mRNA expression of HBx at different time points. MPC5 cells were divided into 4 groups: control group (MPC5 cells cultured under normal conditions), INF-α group (MPC5 cells cultured with INF-α), HBx group (MPC5 cells induced by HBx), and HBx+INF-α group (MPC5 cells induced by HBx and cultured with INF-α). After 48 hours of intervention under different experimental conditions, flow cytometry was used to measure the apoptosis of MPC5 cells, and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of slit diaphragm-related proteins (nephrin, CD2AP, and synaptopodin) and the cytoskeleton-related protein transient receptor potential cation channel 6 (TRPC6).
Results:
MPC5 cells transfected by pEX-HBx had the highest expression of HBx mRNA at 48 hours after transfection (P<0.05). Compared with the control, INF-α and HBx+INF-α groups, the HBx group had a significant increase in the apoptosis rate of MPC5 cells (P<0.05). Compared with the control and INF-α groups, the HBx group had significant reductions in the mRNA and protein expression of nephrin, synaptopodin, and CD2AP and significant increases in the mRNA and protein expression of TRPC6 (P<0.05). Compared with the HBx group, the HBx+INF-α group had significant increases in the mRNA and protein expression of nephrin, synaptopodin, and CD2AP and significant reductions in the mRNA and protein expression of TRPC6 (P<0.05).
Conclusions:
INF-α can inhibit the apoptosis of podocytes induced by HBx, possibly through improving the abnormal expression of slit diaphragm-related proteins (CD2AP, nephrin, and synaptopodin) and cytoskeleton-related protein (TRPC6) induced by HBx.
Insights
Interferon-alfa (INF-α) protects mouse podocytes from hepatitis B virus X (HBx) protein-induced apoptosis. INF-α improves the expression of slit diaphragm proteins and reduces TRPC6, mitigating HBx-induced cellular damage.
Area of Science:
- Molecular biology
- Cell biology
- Virology
Background:
- Hepatitis B virus X (HBx) protein is implicated in liver disease pathogenesis.
- Podocyte injury and apoptosis are key events in kidney disease progression.
- Interferon-alfa (INF-α) is an antiviral cytokine with immunomodulatory effects.
Purpose of the Study:
- To investigate the protective effect of INF-α against HBx-induced podocyte apoptosis.
- To elucidate the molecular mechanisms underlying INF-α's action in podocytes.
Main Methods:
- Mouse podocyte cell line (MPC5) was transfected with HBx gene.
- Cells were treated with INF-α, and apoptosis was assessed by flow cytometry.
- mRNA and protein expression of slit diaphragm proteins (nephrin, CD2AP, synaptopodin) and TRPC6 were analyzed using RT-PCR and Western blot.
Main Results:
- HBx significantly increased MPC5 cell apoptosis.
- HBx induced downregulation of nephrin, CD2AP, and synaptopodin, and upregulation of TRPC6.
- INF-α treatment significantly reduced HBx-induced apoptosis and reversed the abnormal expression of these proteins.
Conclusions:
- INF-α exhibits a protective effect against HBx-induced podocyte apoptosis.
- This protection is mediated by the restoration of slit diaphragm protein expression and modulation of TRPC6.
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