Department of Rheumatology, Allergy and Immunology, Chang Gung Memorial Hospital (at Lin-Kou), Tao-Yuan, Taiwan, ROC; Department of Medicine, Chang Gung University, Tao-Yuan, Taiwan, ROC.
Department of Rheumatology, Allergy and Immunology, Chang Gung Memorial Hospital (at Lin-Kou), Tao-Yuan, Taiwan, ROC.
J Microbiol Immunol Infect. 2020 Feb;53(1):23-32. doi: 10.1016/j.jmii.2017.12.004. Epub 2018 Jan 31.
Renal disease is prevalent in gouty patients and monosodium urate (MSU) crystal deposition in the kidney can be detected in some gouty nephropathy patients. MSU crystals can induce inflammatory events, we investigated the MSU-induced expression of intercellular adhesion molecule (ICAM)-1 on human renal mesangial cells (HRMCs) and the involved signal transduction mechanisms.
The HRMCs cell line was purchased from ScienCell Research Laboratories. MSU crystals were made by dissolving uric acid in sodium hydroxide (NaOH) solution. The involvement of MAPKs, apoptosis-associated speck-like protein containing a CARD domain (ASC), and Toll-like receptor (TLR) was investigated using pharmacological inhibitors, transfection with short hairpin RNA (shRNA), or monoclonal antibodies. Protein expression was evaluated by Western blotting. The functional activity of ICAM-1 was evaluated with cell-cell adhesion assay and immunofluorescence analysis.
MSU stimulation increased expression of ICAM-1 and adhesion between HRMCs and human monocytic THP-1 cells. The interaction between HRMCs and THP-1 was suppressed by ICAM-1 neutralizing antibodies. MSU stimulation induced activation of mitogen-activated protein kinases, including c-Jun N-terminal kinase (JNK), p38, and extracellular signal-regulated kinase (ERK), but only p38 was responsible for MSU-induced expression of ICAM-1 and cell-cell adhesion. ASC also play a role in MSU-induced effects. Pretreatment with monoclonal antibodies against toll-like receptor (TLR)2 or TLR4 reduced MSU-induced ICAM-1 expression, cell-cell adhesion, p38 phosphorylation but the reduction of ASC activation is insignificant.
The MSU induced ICAM-1 expression on HRMCs and cell-cell adhesion involved TLR2/4-p38-ICAM1 pathway and TLR2/4 independent ASC-p38-ICAM1 axis. These findings might partly explain the mechanisms underlying gouty nephropathy.
肾脏疾病在痛风患者中很常见,在一些痛风肾病患者的肾脏中可以检测到单钠尿酸盐(MSU)晶体沉积。MSU 晶体可诱导炎症事件,我们研究了 MSU 诱导的人肾小球系膜细胞(HRMC)细胞间黏附分子(ICAM)-1的表达及其涉及的信号转导机制。
HRMC 细胞系购自 ScienCell 研究实验室。MSU 晶体通过将尿酸溶解在氢氧化钠(NaOH)溶液中制成。使用药理学抑制剂、短发夹 RNA(shRNA)转染或单克隆抗体研究 MAPKs、凋亡相关斑点样蛋白包含 CARD 结构域(ASC)和 Toll 样受体(TLR)的参与情况。通过 Western 印迹评估蛋白质表达。通过细胞-细胞黏附测定和免疫荧光分析评估 ICAM-1 的功能活性。
MSU 刺激增加了 ICAM-1 的表达以及 HRMC 与人类单核细胞 THP-1 细胞之间的黏附。ICAM-1 中和抗体抑制了 HRMC 与 THP-1 之间的相互作用。MSU 刺激诱导丝裂原活化蛋白激酶(包括 c-Jun N 末端激酶(JNK)、p38 和细胞外信号调节激酶(ERK))的激活,但只有 p38 负责 MSU 诱导的 ICAM-1 表达和细胞-细胞黏附。ASC 也在 MSU 诱导的作用中发挥作用。用针对 Toll 样受体(TLR)2 或 TLR4 的单克隆抗体预处理可降低 MSU 诱导的 ICAM-1 表达、细胞-细胞黏附、p38 磷酸化,但 ASC 激活的减少并不明显。
MSU 诱导 HRMC 上的 ICAM-1 表达和细胞-细胞黏附涉及 TLR2/4-p38-ICAM1 途径和 TLR2/4 非依赖性 ASC-p38-ICAM1 轴。这些发现可能部分解释了痛风肾病的发病机制。