目的 研究白花丹素对IgA肾病的作用和机制。 方法 按Ying等改良的BSA+LPS+CCl4方法复制实验IgA肾病模型；然后，给药组大鼠每组分别腹腔注射10 mg/kg、20 mg/kg和50 mg/kg白花丹素，1次/d，对照组和模型组腹腔注射等量的生理盐水1次/d；8周后，全自动化学分析仪检测24 h尿蛋白、血肌酐、血尿素，流式检测ROS含量，试剂盒检测SOD活性和MDA含量，HE染色观察病理损伤，Elisa检测TNF-α、IL-18、IL-1β，蛋白印迹法检测NLRP3、ASC、caspase-1 p20、P13K、AKT和NF-kB蛋白表达。 结果 与模型组相比，给药组大鼠的尿蛋白、血清肌酸酐和尿素氮含量显著减少，ROS水平显著降低，SOD含量显著增多，MDA含量显著减少，MDA、IL-1β、IL-18和TNF-α的含量显著减少，NLRP3、ASC、caspase-1 p20、P13K、AKT和NF-kB的蛋白表达显著下调，并且随着给药量的增加效果越显著。 结论 白花丹素通过降低尿蛋白、血肌酐和血尿素含量，减轻病理损伤，抑制氧化应激、炎症反应和NLRP3/P13K/AKT/NF-kB通路激活来减轻IgA肾病。
Objective To investigate the role and mechanism of plumbagin in IgA nephropathy. Methods First, the IgA nephropathy model was constructed according to the modified BSA+LPS+CCl4 method of Ying et al. Then, the rats in the administration group were intraperitoneally injected with 10 mg/kg, 20 mg/kg or 50 mg/kg of plumbagin per day. The normal group and the model group were intraperitoneally injected with the same amount of normal saline per day, after 8 weeks later, 24 h urine protein, serum creatinine, and blood urea were detected by a fully automatic chemical analyzer, and the ROS content was detected by flow, and the SOD activity and MDA content were detected. Pathological damage was observed by HE staining, TNF-a, IL-18, IL-1β were detected by Elisa, and NLRP3, ASC, caspase-1 p20, P13K, protein expression of AKT and NF-kB were detected by Western blotting. Results Compared with the model group, the proteinuria, serum creatinine and urea nitrogen levels of the administration group significantly reduced, ROS levels significantly decreased, SOD levels increased, MDA, IL-1b, IL-18 and TNF-a levels were significantly increased. Protein expression of NLRP3, ASC, caspase-1 p20, P13K, AKT and NF-kB was significantly down-regulated, with the increasing of dose, the effect was more pronounced. Conclusions Plumbagin mainly reduces IgA nephropathy by reducing urinary protein, serum creatinine and blood urea, reducing pathological damage, inhibiting oxidative stress, inflammatory response and activation of NLRP3 / P13K / AKT / NF-kB pathway.
[1] Bollin R, Haller H. Pathophysiology and treatment of IgA nephropathy[J]. Internist, 2018, 59(7): 736-740.
[2] Xue YL, Meng XQ, Ma LJ, et al. Plumbagin exhibits an anti-proliferative effect in human osteosarcoma cells by downregulating FHL2 and interfering with Wnt/β-catenin signalling[J]. Oncol Lett, 2016, 12(2): 1095-1100.
[3] Yong R, Chen XM, Shen S, et al. Plumbagin ameliorates diabetic nephropathy via interruption of pathways that include NOX4 signalling[J]. PloS One, 2013, 8(8): e73428.
[4] Nakahira K, Haspel JA, Rathinam VA, et al. Autophagy proteins regulate innate immune responses by inhibiting the release of mitochondrial DNA mediated by the NPLP3 inflammasome[J]. Nat Immunol, 2011, 12(3): 222-230.
[5] Tsai YL, Hua KF, Chen A, et al. Nlrp3 inflammasome: pathogenic role and potential therapeutic target for IgA nephropathy[J]. Sci Rep, 2017, 7: 41123.
[6] Ying T , Tan-Qi L , Cai-Lian C , et al. Improvement of experimental IgA nephropathy model[J]. J Sun Yat-Sen University(Medical Sciences), 2006, 27(2): 184-187.
[7] 王巍巍, 陈以平. 黄芪牛蒡子不同配伍对STZ大鼠肾组织内活性氧产物含量和核转录因子-kB表达的影响[J]. 中国中西医结合杂志, 2008, 28(10): 917-920.
[8] Chu H, Yu H, Ren D, et al. Plumbagin exerts protective effects in nucleus pulposus cells by attenuating hydrogen peroxide-induced oxidative stress, inflammation and apoptosis through NF-kB and Nrf-2[J]. Int J Mol Med, 2016, 37(6): 1669-1676.
[9] Rauen T, Floege J. Inflammation in IgA nephropathy[J]. Pediatr Nephrol (Berlin, Germany), 2017, 32(12): 2215-2224.
[10]Zheng XY, Mao CY, Qiao H, et al. Plumbagin suppresses chronic periodontitis in rats via down-regulation of TNF-α, IL-1β and IL-6 expression[J]. Acta Pharmacol Sin, 2017, 38(8): 1150-1160.
[11] Messeha SS, Zarmouh NO, Mendonca P, et al. The attenuating effects of plumbagin on pro-inflammatory cytokine expression in LPS-activated BV-2 microglial cells[J]. J Neuroimmunol, 2017, 313: 129-137.
[12]Gupta AC, Mohanty S, Saxena A, et al. Plumbagin, a vitamin K3 analogue ameliorate malaria pathogenesis by inhibiting oxidative stress and inflammation[J]. Inflammopharmacology, 2018, 26(4): 983-991.
[13] Hua KF, Yang SM, Kao TY, et al. Osthole mitigates progressive IgA nephropathy by inhibiting reactive oxygen species generation and NF-kB/NLRP3 pathway[J]. PloS One, 2013, 8(10): e77794.
[14]Wu H, Dai X, Wang E. Plumbagin inhibits cell proliferation and promotes apoptosis in multiple myeloma cells through inhibition of the PI3K/Akt-mTOR pathway[J]. Oncol Lett, 2016, 12(5): 3614-3618.
京ICP备08002354号-3
