小儿紫癜疹消颗粒对过敏性紫癜患儿血清IgA1诱导人脐静脉内皮细胞炎症的抑制机制研究
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摘要
目的探讨小儿紫癜疹消颗粒对过敏性紫癜(HSP)患儿血清IgA1诱导人脐静脉内皮细胞(HUVECs)炎症的抑制作用,并初步阐明其作用机制。方法以HUVECs为研究对象,用健康儿童血清IgA1和HSP患儿血清IgA1进行干预,同时加入不同浓度的小儿紫癜疹消颗粒,分为空白对照组、正常对照组、HSP组及低、中、高浓度中药组(25、50、100 mg/L),各组于培养24 h后,采用酶联免疫吸附法测定各组上清液中肿瘤坏死因子α(TNF-α)、白细胞介素-8(IL-8)水平。Westernbolt检测各组细胞内核因子κB(NF-κB)蛋白表达水平。结果各组细胞上清液中TNF-α、IL-8水平,与空白对照组和正常对照组相比,HSP组显著升高,差异有统计学意义(P<0.001);与HSP组比较,低、中、高浓度中药组TNF-α、IL-8水平明显降低,差异有统计学意义(P<0.05)。Western blot检测各组HUVECs P65蛋白表达水平,与空白对照组和正常对照组比较,HSP组P65蛋白显著升高,差异有统计学意义(P<0.05);与HSP组比较,高浓度中药组P65蛋白水平明显降低,差异有统计学意义(P<0.05)。结论小儿紫癜疹消颗粒可使HUVECs中NF-κB信号通路P65蛋白的磷酸化水平降低,减少相关炎症因子TNF-α、IL-8的分泌,从而有效抑制细胞炎症反应的发生。
Objective To investigate the inhibitory effect of Xiao'er Zidian Zhenxiao granules on the inflammation of human umbilical venous endothelial cells(HUVECs) induced by serum IgA1 in children with Henoch-Sch?nlein purpura(HSP), and to clarify its mechanism.Methods HUVECs were used as the research subjects, serum IgA1 in HSP children and in healthy children were used to intervene, and different concentrations of Xiao'er Zidian Zhenxiao granules were added. They were divided into blank control group, normal control group, Henoch-Sch?nlein purpura group(HSP group) and the Xiao'er Zidian Zhenxiao granules groups(ZXKL groups)(the low-,medium-and high-concentration group, with the concentration being 25 mg/L, 50 mg/L and 100 mg/L respectively). After 24 hours of culturing in each group, TNF-α and IL-8 levels in the supernatant of each group were determined by enzyme-linked immunosorbent assay. Westernbolt was used to detect the expression of NF-κB protein in each group.Results The levels of TNF-α and IL-8 in the supernatant of each group were significantly higher in the HSP group than in the blank and normal control groups(P<0.001). Compared with the HSP group, the levels of TNF-α and IL-8 in the ZXKL groups decreased(P<0.05). The expression of P65 protein in HUVECs was detected by Western blot. Compared with the blank and normal control groups, the P65 protein level in the HSP group was significantly increased(P<0.05). Compared with the HSP group, the P65 protein level in the high-concentration ZXKL group(100 mg/L) was decreased(P<0.05).Conclusion Xiao'er Zidian Zhenxiao granules can decrease the phosphorylation of NF-κB signaling pathway P65 protein in HUVECs and reduce the secretion of related inflammatory cytokines(TNF-α and IL-8), thus effectively inhibiting the occurrence of cellular inflammatory responses.
Objective To investigate the inhibitory effect of Xiao'er Zidian Zhenxiao granules on the inflammation of human umbilical venous endothelial cells(HUVECs) induced by serum IgA1 in children with Henoch-Sch?nlein purpura(HSP), and to clarify its mechanism.Methods HUVECs were used as the research subjects, serum IgA1 in HSP children and in healthy children were used to intervene, and different concentrations of Xiao'er Zidian Zhenxiao granules were added. They were divided into blank control group, normal control group, Henoch-Sch?nlein purpura group(HSP group) and the Xiao'er Zidian Zhenxiao granules groups(ZXKL groups)(the low-,medium-and high-concentration group, with the concentration being 25 mg/L, 50 mg/L and 100 mg/L respectively). After 24 hours of culturing in each group, TNF-α and IL-8 levels in the supernatant of each group were determined by enzyme-linked immunosorbent assay. Westernbolt was used to detect the expression of NF-κB protein in each group.Results The levels of TNF-α and IL-8 in the supernatant of each group were significantly higher in the HSP group than in the blank and normal control groups(P<0.001). Compared with the HSP group, the levels of TNF-α and IL-8 in the ZXKL groups decreased(P<0.05). The expression of P65 protein in HUVECs was detected by Western blot. Compared with the blank and normal control groups, the P65 protein level in the HSP group was significantly increased(P<0.05). Compared with the HSP group, the P65 protein level in the high-concentration ZXKL group(100 mg/L) was decreased(P<0.05).Conclusion Xiao'er Zidian Zhenxiao granules can decrease the phosphorylation of NF-κB signaling pathway P65 protein in HUVECs and reduce the secretion of related inflammatory cytokines(TNF-α and IL-8), thus effectively inhibiting the occurrence of cellular inflammatory responses.
引文
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[19] 周平,罗云,邢娜,等.肿瘤坏死因子 α 介导动脉粥样硬化发生机制的研究进展[J].世界中医药,2015,10(8) :1163-1168.
[20] Tay C,Liu YH,Hosseini H,et al.B-cell-specific depletion of tumour necrosis factor alpha inhibits atherosclerosis developmentand plaque vulnerability to rupture by reducing cell death and inflammation[J].Cardiovasc Res,2016,111(4):385-397.
[21] 陈铁龙,祝光礼,张旭栋,等.TNF-α诱导人脐静脉血管内皮细胞凋亡和炎症机制研究[J].中华全科医学,2018,16(2):184-187.
[22] 华冉,鹿玲.过敏性紫癜患儿血清低糖基化IgA1、核因子κB与炎症递质的研究[J].临床儿科杂志,2016,34(4):268-272.
[23] Hayden MS,Ghosh S.Shared principles in NF-kappaB signaling[J].Cell,2008,132(3):344-362.
[24] Gasparini C,Feldmann M.NF-κB as a target for modulating inflammatory responses[J].Curr Pharm Des,2012,18(35):5735-5745.
[2] Kiryluk K,Moldoveanu Z,Sanders JT,et al.Aberrant glycosylation of IgA1 is inherited in both pediatric IgA nephropathy and Henoch-Sch?nlein purpura nephritis[J].Kidney Int,2011,80(1):79-87.
[3] Davin JC,Coppo R.Henoch-Sch?nlein purpura nephritis in children[J].Nat Rev Nephrol,2014,10(10):563-573.
[4] 吴琳,袁丽萍,费文君,等.过敏性紫癜患儿血清对人脐静脉内皮细胞的影响及甲基泼尼松龙的保护作用[J].中国当代儿科杂志,2012,14(1):59-63.
[5] 朱浩宇,刘晓蕾,冯晓纯.小儿紫癜疹消颗粒治疗过敏性紫癜(风热伤络型)40例临床观察[J].中国卫生标准管理,2016,7(8):123-125.
[6] Ozen S,Ruperto N,Dillon MJ,et al.EULAR/PReS endorsed consensus criteria for the classification of childhood vasculitides[J].Ann Rheum Dis,2006,65(7):936-941.
[7] Glynn KM,Anderson P,Fast DJ,et al.Gromwell (Lithospermum erythrorhizon) root extract protects against glycation and relatedinflammatory and oxidative stress while offering UV absorption capability[J].Exp Dermatol,2018,27(9):1043-1047.
[8] Shen CH,Liu CT,Song XJ,et al.Evaluation of analgesic and anti-inflammatory activities of Rubia cordifolia L.by spectrum-effectrelationships[J].J Chromatogr B Analyt Technol Biomed Life Sci,2018,1090:73-80.
[9] Ma T,Wang Z,Zhang YM,et al.Bioassay-Guided Isolation of Anti-Inflammatory Components from the Bulbs of Lilium browniivar viridulum and Identifying the Underlying Mechanism through Acting on the NF-κB/MAPKs Pathway[J].Molecules,2017,22(4):e506.
[10] 江国荣,刘少文,陈卫民,等.不同炮制方法对牡丹皮中有效成分含量的影响[J].临床合理用药杂志,2017,10(28):15-16.
[11] Choi YY,Kim MH,Lee H,et al.Cynanchum atratum inhibits the development of atopic dermatitis in 2,4-dinitrochlorobenzene-induced mice[J].Biomed Pharmacother,2017,90:321-327.
[12] Han T,Li J,Xue J,et al.Scutellarin derivatives as apoptosis inducers:Design,synthesis and biological evaluation[J].Eur J Med Chem,2017,135:270-281.
[13] 刘雷,郭丽娜,于春磊,等.白鲜皮化学成分及药理活性研究进展[J].中成药,2016,38(12):2657-2665.
[14] 张俊飞,孙广璐,张彬,等.侧柏叶药理作用的研究进展[J].时珍国医国药,2013,24(9):2231-2233.
[15] Lundstedt AC,McCarthy S,Gustafsson MC,et al.A genetic basis of susceptibility to acute pyelonephritis[J].PLoS One,2007,2(9):e825.
[16] Lundstedt AC,Leijonhufvud I,Ragnarsdottir B,et al.Inherited susceptibility to acute pyelonephritis:a family study of urinary tract infection[J].J Infect Dis,2007,195(8):1227-1234.
[17] Ragnarsdóttir B,Svanborg C.Susceptibility to acute pyelonephritis or asymptomatic bacteriuria:host-pathogen interaction in urinary tract infections[J].Pediatr Nephrol,2012,27(11):2017-2029.
[18] Yang YH,Lai HJ,Huang CM,et al.Sera from children with active Henoch-Sch?nlein purpura can enhance the production of interleukin 8 by human umbilical venous endothelial cells[J].Ann Rheum Dis,2004,63(11):1511-1513.
[19] 周平,罗云,邢娜,等.肿瘤坏死因子 α 介导动脉粥样硬化发生机制的研究进展[J].世界中医药,2015,10(8) :1163-1168.
[20] Tay C,Liu YH,Hosseini H,et al.B-cell-specific depletion of tumour necrosis factor alpha inhibits atherosclerosis developmentand plaque vulnerability to rupture by reducing cell death and inflammation[J].Cardiovasc Res,2016,111(4):385-397.
[21] 陈铁龙,祝光礼,张旭栋,等.TNF-α诱导人脐静脉血管内皮细胞凋亡和炎症机制研究[J].中华全科医学,2018,16(2):184-187.
[22] 华冉,鹿玲.过敏性紫癜患儿血清低糖基化IgA1、核因子κB与炎症递质的研究[J].临床儿科杂志,2016,34(4):268-272.
[23] Hayden MS,Ghosh S.Shared principles in NF-kappaB signaling[J].Cell,2008,132(3):344-362.
[24] Gasparini C,Feldmann M.NF-κB as a target for modulating inflammatory responses[J].Curr Pharm Des,2012,18(35):5735-5745.