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Protective effect of Qingre Zhixue Formula combined with tripterygium wilfordii multiglucoside on endothelial injury induced by serum-derived poly IgA in Henoch-Schönlein purpura nephritis and effects on the NF-κB pathway

  • role： First author第一作者
  • Affiliation：

    The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450003, China

    School of Pediatrics, Henan University of Chinese Medicine, Zhengzhou 450046, China

  • Introduction：张霞，女，博士，主任医师
ZHANG Xia12,  
  • Affiliation：

    The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450003, China

    School of Pediatrics, Henan University of Chinese Medicine, Zhengzhou 450046, China

XU Shanshan12,  
  • Affiliation：

    The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450003, China

    School of Pediatrics, Henan University of Chinese Medicine, Zhengzhou 450046, China

WANG Long12,  
  • Affiliation：

    Xuchang Hospital of Chinese Medicine, Xuchang 461099, China

SUN Yang3,  
  • Affiliation：

    The Affiliated Hospital of Hangzhou Normal University, Hangzhou 310018, China

DI Jiaqi4,  
  • Affiliation：

    School of Pediatrics, Henan University of Chinese Medicine, Zhengzhou 450046, China

DAI Yanlin2,  
  • role： Corresponding author通信作者
  • Affiliation：

    The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450003, China

    School of Pediatrics, Henan University of Chinese Medicine, Zhengzhou 450046, China

  • Email：dingying3236@sina.com
  • Introduction：Prof. DING Ying, Master of Chinese Medicine, Chief Physician, Doctoral Supervisor.The First Affiliated Hospital of Henan University of Chinese Medicine, No.19, Renmin Road, Jinshui District, Zhengzhou 450003.E-mail: dingying3236@sina.com
DING Ying12*

Resümee

ObjectiveWe aimed to explore the protective effect of Qingre Zhixue Formula and tripterygium wilfordii multiglucoside on the endothelial damage induced by serum-derived poly IgA in patients with Henoch-Schönlein purpura nephritis and its effect on the expression of key proteins of the NF-κB signaling pathway.MethodsA total of 80 patients with Henoch-Schönlein purpura nephritis from The First Affliated Hospital of Henan University of Chinese Medicine and 38 healthy controls were enrolled in this study. Serum IgA was purified from all participants in order to prepare serum-derived poly IgA. Moreover, 60 Sprague-Dawley rats were divided into the following five groups (n=12 rats per group): the Qingre Zhixue Formula group (31.25 mg/kg), the tripterygium wilfordii multiglucoside group (46.88 mg/kg), the combined group (31.25 mg/kg + 46.88 mg/kg), the prednisone acetate group (23.44 mg/kg) and the saline group. Rats in each group were gavaged twice a day for 7 days. Additionally, human umbilical vein endothelial cells (HUVECs) were stimulated with serum-derived poly IgA (200 μg/mL) derived from patients with Henoch-Schönlein purpura nephritis for 24 h to construct HUVECs injury model. The cells were divided into the following seven groups: the blank group, the healthy group, the model group, the Qingre Zhixue Formula group, the tripterygium wilfordii multiglucoside group, the combined group and the prednisone acetate group, and provide corresponding intervention. siRNA transfection was used to inhibit the NF-κB signaling pathway, and divide the transfected cells into the blank group, the model group, the model+ normal control group, the model+ transfection group, the Qingre Zhixue Formula group, the tripterygium wilfordii multiglucoside group, the combined group and the prednisone acetate group. Cell viability was detected by the MTT assay, the apoptosis rate was detected by flow cytometry, and the protein expression levels of p-P65, p-P50, IKKβ, and IκBα were detected by Western blotting before and after transfection.ResultsCompared with the blank group, the cell viability of the model group was decreased (P<0.05). Compared with the model group, the cell viability of each treatment group was increased (P<0.05). Compared with the combined group, the cell viability of the Qingre Zhixue Formula group, the tripterygium wilfordii multiglucoside group, and the prednisone acetate group was decreased (P<0.05). Compared with the blank group, the protein expression levels of p-P65, p-P50 and IKKβ in the model group were increased (P<0.05), and the protein expression level of IκBα was decreased (P<0.05). Compared with the model group, the protein expression levels of p-P65, p-P50 and IKKβ were decreased in each group (P<0.05), and the protein expression level of IκBα was increased (P<0.05). Compared with the combined group, the protein expression levels of p-P65, p-P50, and IKKβ in the Qingre Zhixue Formula group, the tripterygium wilfordii multiglucoside group and the prednisone acetate group were increased (P<0.05), while the protein expression level of IκBα was decreased (P<0.05). After siRNA transfection, compared with the blank group, the protein expression levels of p-P65, p-P50 and IKKβ in the model group and the model+ normal control group were increased (P<0.05), and the protein expression level of IκBα was decreased (P<0.05). Compared with the model group, the protein expression levels of p-P65, p-P50 and IKKβ were decreased (P<0.05), and the protein expression level of IκBα was increased (P<0.05) in the cells of the model+ transfection group, the Qingre Zhixue Formula group, the tripterygium wilfordii multiglucoside group, the combined group and the prednisone acetate group.ConclusionQingre Zhixue Formula combined with tripterygium wilfordii multiglucoside may inhibit the activation of the NF-κB pathway to improve the cell viability of HUVECs induced by serum-derived poly IgA in children with Henoch-Schönlein purpura nephritis, so it plays a role in protecting endothelial cells.

Schlüsselwort

Henoch-Schönlein purpura nephritis;Qingre Zhixue Formula;tripterygium wilfordii multiglucoside;human umbilical vein endothelial cells;NF-κB pathway

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