Effect of triptolide on the regulation of inflammatory response and apoptosis in PBMC from IgA nephropathy patients
WANG Chang, LIANG Qian, PENG You-ming, LIU Hong, SUN Lin, YIN Wen-qing, LING Guang-hui, LIU Fu-you.
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Department of Nephrology, Key Laboratory of Nephrology and Blood Purification in Hunan, the Second Xiangya Hospital, Central South University, Changsha 410011, China Corresponding author: PENG You-ming, Email: youmingpeng5577@hotmail.com
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History+
Received
Revised
Published
1900-01-01
1900-01-01
2011-10-15
Issue Date
2011-10-15
Abstract
Objective To investigate the inflammatory response and apoptosis of peripheral blood mononuclear cells (PBMCs) and their regulation by triptolide (TP) in IgA nephropathy (IgAN) patients. Methods Blood samples were collected from 29 IgAN patients and 16 healthy individuals. TNF-α and IL-6 concentrations were measured by ELISA and NO concentration by Griess reagent in the plasma of samples. PBMCs were isolated from IgAN patients and cultured in vitro, and subsequently activated by PHA (10 mg/L). The cytotoxicity of different TP concentrations was assayed by MTT and two non-toxic concentrations (12.5 μg/L or 25.0 μg/L) were selected for treatment. TNF-α, IL-6 and NO concentrations were measured in the culture media collected from PBMCs cultures activated by PHA (10 mg/L) and treated with TP (12.5 μg/L or 25.0 μg/L). The PHA-activated, TP-treated cells apoptotic rate was analyzed by FACS using Annexin V-FITC staining. The expression of Bcl-2, Bax, caspase-9 and caspase-3 were detected by RT-PCR and Western blotting from lyses of PHA-activated with or without TP-treated cells. Results The serum concentrations of TNF-α [(131.57±50.61) ng/L vs (30.24±18.93) ng/L, P<0.01], IL-6[(76.36±25.21) ng/L vs(35.08±16.59) ng/L, P<0.01] and NO[(46.36±12.93) μmol/L vs(26.61±10.87) μmol/L, P<0.01] were significantly increased in IgAN patients compared to healthy individuals. PBMCs viability in culture decreased after TP treatment in a dose-dependent manner. TP also inhibited TNF-α, IL-6 and NO levels in the media of PHA-activated PBMCs in culture and induced PBMCs apoptosis. The expression of Bcl-2 decreased markedly and Bax, caspase-9 and caspase-3 increased significantly after TP treatment（all P<0.05). Conclusions The PBMCs from IgAN patients are in a highly activated state, and have a high apoptotic rate. TP treatment induces benificial effects in IgAN patients by inhibiting the activation of PBMCs by activating pro-apoptotic pathway.
WANG Chang;LIANG Qian;PENG You-ming;LIU Hong;SUN Lin;YIN Wen-qing;LING Guang-hui;LIU Fu-you. .
Effect of triptolide on the regulation of inflammatory response and apoptosis in PBMC from IgA nephropathy patients[J]. Chinese Journal of Nephrology, 2011, 27(10): 724-729.
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