[Protective Effect of Total Flavones of Bidens pilosa L. on IgA1 Induced Injury of HUVECs in Henoch-Schönlein Purpura Children Patients].

OBJECTIVE: To explore the protective effect and mechanism of total flavones of Bidens pilosa L. (TFB) on IgA1 induced injury of venous endothelial cells in Henoch-Schönlein purpura (HSP) children patients. METHODS Human umbilical venous endothelial cells (HUVECs) were taken as subject. They were intervened by normal IgA1 and HSP children patients' serum IgA1, and added with different concentrations TFB at the same time. Then they were divided into the blank control group, the normal control group, the HSP IgA1 group, and HSP IgA1 plus TFB (1.0, 0.5, 0.25 mg/mL) groups. Levels of TNF-α and IL-8 in supernate were detected by ELISA. The NO level was detected by nitrate reductase method. mRNA and protein expressions of NF-κB and ICAM-1 in HUVECs were detected by fluorescent quantitative PCR and Western blot respectively. RESULTS: Compared with the normal control group and the blank control group, levels of IL-8, TNF-α, and NO all significantly increased in the HSP group (P < 0.05). Compared with the HSP group, levels of IL-8, TNF-α, and NO significantly decreased after intervention of TFB (1.0 and 0.5 mg/mL; P < 0.05, P < 0.01). Results of fluorescent quantitative PCR and Western blot showed, as compared with the blank control group and the normal control group, mRNA and protein expressions of NF-κB and ICAM-1 in HSP children patients' serum IgA1 induced venous endothelial cells significantly increased with statistical difference (P < 0.05, P < 0.01). Compared with the HSP group, mRNA and protein expressions of NF-KB and ICAM-1 were obviously down-regulated after intervention of TFB (1.0, 0.5, 0.25 mg/mL), with statistical difference (P < 0.05, P < 0.01). CONCLUSION: TFB could protect vascular damage by inhibiting in vivo high expression of NF-κB, reducing the production of IL-8, TNF-α, and NO in vascular endothelial cells of HSP children patients.

[Apoptosis of human umbilical vein endothelial cells induced by IgA1 isolated from Henoch-Schönlein purpura patients].

OBJECTIVE: To observe the effect of apoptosis of human umbilical vein endothelial cells (HUVEC) induced by IgA1 from Henoch-Schönlein purpura (HSP) patients. METHOD: HUVEC were cultured in 3 different conditional media with IgA1 from HSP patients, normal healthy children and simply the cell culture medium. Serum IgA1 was purified by jacalin affinity chromatography, rates of apoptosis in HUVEC cells at different concentration and different times after incubation with IgA1 were determined by TUNEL method and flow cytometry. Real-time PCR and Western blot methods were used to detect the expression of caspase-3 and Fas, respectively. RESULT: Apoptosis rate of HUVEC by IgA1 isolated from HSP patients were significantly higher than that of the blank control [(14.77 ± 2.23)% vs. (2.25 ± 0.77)%, P < 0.01] and the apoptosis rate of HUVEC induced by IgA1 from normal healthy children was higher than that of blank control [(7.97 ± 1.48)% vs. (2.25 ± 0.77)%, P < 0.01]. The apoptosis rate of HUVEC induced by IgA1 from HSP was time and concentration-dependent. Moreover IgA1 isolated from HSP patients could significantly increase the caspase-3 and Fas expression (P < 0.01). CONCLUSION: The IgA1 from HSP patients could induce the apoptosis of HUVEC, which might be related to the progression of HSP.

[Impact of sera from children with active Henoch-Schönlein purpura on human umbilical venous endothelial cells (HUVECs) and protective effects of methylprednisolone against HUVECs injury].

OBJECTIVE: To observe the changes of human umbilical venous endothelial cells (HUVECs) induced by the sera from children with active Henoch-Sch-nlein purpura (HSP) and the protective effects of methylprednisolone against HUVECs injury. METHODS: HUVECs were divided into four groups based on the culture conditions: blank control group, normal serum group, HSP serum group, and HSP serum plus methylprednisolone group. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-8 in the supernatants of each group were detected using ELISA and the nitric oxide (NO) level by nitrate reductase determination. Moreover, the expressions of nuclear factor-kappa B (NF-κB) and Fractalkine in HUVECs were examined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. RESULTS: The levels of IL-8, TNF-α, and NO in the HSP serum group were significantly higher than those in the blank control and normal serum groups (P<0.05). Compared with the HSP serum group, the levels of IL-8, TNF-α, and NO in the HSP serum plus methylprednisolone group decreased significantly (P<0.05). The mRNA expression levels of NF-κB and Fractalkine in the HSP serum group were significantly higher than those in the blank control group (P<0.05). The protein expression levels of NF-κB and Fractalkine in the HSP serum group were significantly higher than those in the blank control and normal control group (P<0.05). Compared with the HSP serum group, the mRNA and protein expression levels of NF-κB and Fractalkine in the HSP serum plus methylprednisolone group decreased significantly (P<0.05). CONCLUSIONS: The sera from children with active HSP can induce the in vitro cultured HUVECs to become activated and excrete cytokines. Methylprednisolone may inhibit NF-κB expression, reduce the production of inflammatory factors, and thus alleviate vascular inflamation.