Dinesh Palani, Rasool MahaboobKhan
Immunopathology Lab, School of Bio Sciences and Technology, VIT University, Vellore 632 014, Tamil Nadu, India.
Immunopathology Lab, School of Bio Sciences and Technology, VIT University, Vellore 632 014, Tamil Nadu, India.
Int Immunopharmacol. 2017 Mar;44:26-37. doi: 10.1016/j.intimp.2016.12.031. Epub 2017 Jan 7.
The current study was designed to investigate the therapeutic potential of berberine on monosodium urate (MSU) crystal stimulated RAW 264.7 macrophages and in MSU crystal induced rats. Our results indicate that berberine (25, 50 and 75μM) suppressed the levels of pro-inflammatory cytokines (interleukin-1beta (IL-1β) and tumor necrosis factor alpha (TNFα)) and intracellular reactive oxygen species in MSU crystal stimulated RAW 264.7 macrophages. The mRNA expression levels of IL-1β, caspase 1, nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 (NLRP3), thioredoxin interacting protein (TXNIP) and kelch-like ECH-associated protein 1 (Keap1) were found downregulated with the upregulation of nuclear factor erythroid-2-related factor 2 (Nrf2) transcription factor and its associated anti-oxidant enzymes: Heme oxygenase I (HO-1), superoxide dismutase (SOD1), glutathione peroxidase (GPx), NADPH quinone oxidoreductase-1 (NQO1) and catalase (CAT) in MSU crystal stimulated RAW 264.7 macrophages upon berberine treatment. Subsequently, western blot analysis revealed that berberine decreased the protein expression of IL-1β and caspase 1 and increased Nrf2 expression in RAW 264.7 macrophages. Immunofluorescence analysis also explored increased expression of Nrf2 in MSU crystal stimulated RAW 264.7 macrophages by berberine treatment. In addition, the paw edema, pain score, pro-inflammatory cytokines (IL-1β and TNFα) and articular elastase activity were found significantly reduced in berberine (50mg/kgb·wt) administered MSU crystal-induced rats. Conclusively, our current findings suggest that berberine may represent as a potential candidate for the treatment of gouty arthritis by suppressing inflammatory mediators and activating Nrf2 anti-oxidant pathway.
本研究旨在探讨小檗碱对尿酸钠（MSU）晶体刺激的RAW 264.7巨噬细胞以及MSU晶体诱导的大鼠的治疗潜力。我们的结果表明，小檗碱（25、50和75μM）可抑制MSU晶体刺激的RAW 264.7巨噬细胞中促炎细胞因子（白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNFα））的水平以及细胞内活性氧的水平。研究发现，在小檗碱处理后，MSU晶体刺激的RAW 264.7巨噬细胞中IL-1β、半胱天冬酶1、含pyrin结构域的核苷酸结合寡聚化结构域样受体3（NLRP3）、硫氧还蛋白相互作用蛋白（TXNIP）和kelch样ECH相关蛋白1（Keap1）的mRNA表达水平下调，而核因子红细胞2相关因子2（Nrf2）转录因子及其相关抗氧化酶血红素加氧酶I（HO-1）、超氧化物歧化酶（SOD1）、谷胱甘肽过氧化物酶（GPx）、NADPH醌氧化还原酶-1（NQO1）和过氧化氢酶（CAT）的表达上调。随后，蛋白质印迹分析显示，小檗碱可降低RAW 264.7巨噬细胞中IL-1β和半胱天冬酶1的蛋白表达，并增加Nrf2的表达。免疫荧光分析还发现，小檗碱处理可使MSU晶体刺激的RAW 264.7巨噬细胞中Nrf2的表达增加。此外，在给予小檗碱（50mg/kg体重）的MSU晶体诱导大鼠中，发现爪部水肿、疼痛评分、促炎细胞因子（IL-1β和TNFα）以及关节弹性蛋白酶活性均显著降低。总之，我们目前的研究结果表明，小檗碱可能通过抑制炎症介质和激活Nrf2抗氧化途径，成为治疗痛风性关节炎的潜在候选药物。
