Efficacy of triptolide on the apoptosis of tonsillar mononuclear cells from patients with IgA nephropathy
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Updated: Apr 1, 2026
Isolation of Tonsillar Mononuclear Cells to Study Ex Vivo Innate Immune Responses in a Human Mucosal Lymphoid Tissue
Published on: June 14, 2020
Wenzhe Yan1, Youming Peng1, Chan Liu1
1a Department of Nephrology , The Second Xiangya Hospital, Central South University, Hunan Key Lab of Kidney Disease and Blood Purification , People's Republic of China and.
Aims:
This study aimed to evaluate the extent of apoptosis of tonsillar mononuclear cells (TMCs) derived from patients with IgA nephropathy (IgAN) and the effects of triptolide (TP) on the apoptosis of these TMCs.
Methods:
TMCs were isolated from tonsillar tissues of patients with IgAN or chronic tonsillitis (control group). Rates of TMCs apoptosis were measured by annexin V-fluorescein isocyanate (FITC)/propidium iodide (PI)-labeled flow cytometry (FCM). Expression levels of Bcl-2 family proteins were quantified by immunohistochemistry of fixed tonsillar sections and Western blot analyzes of TMCs lysates. TMCs from IgAN patients were treated 10, 20, or 30 ng/mL TP for 24 h and then evaluated for viability by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, for the percentage of apoptotic cells by FCM, and for the relative expression levels of Bcl-2 family proteins by Western blot analysis.
Results:
Compared to TMCs from the control group, TMCs from the IgAN group demonstrated lower rates of apoptosis, higher expression levels of the anti-apoptosis proteins Bcl-2 and Bcl-xL, and lower expression levels of the pro-apoptosis protein Bax. Treatment of IgAN patient-derived TMCs with 10, 20, or 30 ng/mL TP for 24 h suppressed the viability and promoted the apoptosis of TMCs in a dose-dependent manner. Western blot analysis revealed a TP dose-dependent decrease in Bcl-2 and Bcl-xL expression levels, in parallel with increased Bax protein levels.
Conclusion:
TMCs from IgAN patients may be in a state of inhibited apoptosis mediated by Bcl-2 family proteins, which may be reversed by TP treatment.
Insights
Tonsillar mononuclear cells (TMCs) from IgA nephropathy (IgAN) patients show reduced apoptosis, potentially due to Bcl-2 family proteins. Triptolide (TP) treatment reversed this inhibition, promoting apoptosis in IgAN patient-derived TMCs.
Area of Science:
- Immunology
- Cell Biology
- Nephrology
Background:
- IgA nephropathy (IgAN) is a kidney disease characterized by immune system dysregulation.
- Tonsillar mononuclear cells (TMCs) play a role in the immune response and may be involved in IgAN pathogenesis.
- Apoptosis, or programmed cell death, is a critical cellular process that can be dysregulated in disease states.
Purpose of the Study:
- To investigate the rate of apoptosis in TMCs from IgAN patients compared to controls.
- To examine the expression levels of Bcl-2 family proteins, which regulate apoptosis, in IgAN patient-derived TMCs.
- To evaluate the effect of triptolide (TP), a natural compound, on TMC apoptosis and Bcl-2 family protein expression in IgAN.
Main Methods:
- TMCs were isolated from tonsils of IgAN patients and healthy controls (chronic tonsillitis).
- Apoptosis rates were measured using flow cytometry (FCM) with annexin V-FITC/propidium iodide staining.
- Bcl-2 family protein expression was assessed via immunohistochemistry and Western blot analysis.
- IgAN patient-derived TMCs were treated with varying concentrations of triptolide (TP) and analyzed for viability (MTT assay) and apoptosis (FCM).
Main Results:
- TMCs from IgAN patients exhibited significantly lower apoptosis rates compared to controls.
- IgAN patient-derived TMCs showed increased expression of anti-apoptotic proteins (Bcl-2, Bcl-xL) and decreased expression of pro-apoptotic protein (Bax).
- TP treatment dose-dependently reduced TMC viability and increased apoptosis in IgAN patient-derived cells.
- TP treatment led to decreased Bcl-2 and Bcl-xL expression and increased Bax protein levels in a dose-dependent manner.
Conclusions:
- TMCs from IgAN patients may undergo inhibited apoptosis, potentially mediated by altered Bcl-2 family protein expression.
- Triptolide (TP) demonstrates a potential therapeutic effect by reversing this apoptotic inhibition in IgAN patient-derived TMCs.
- These findings suggest a novel therapeutic strategy targeting apoptosis in IgAN.
