Silswal Neerupma, Reis Julia, Qureshi Asaf A, Papasian Christopher, Qureshi Nilofer
*Department of Basic Medical Science, School of Medicine, and Shock/Trauma Research Center, School of Medicine, Kansas City, Missouri †Department of Pharmacology and Toxicology, School of Pharmacy, University of Missouri Kansas City, Kansas City, Missouri.
Shock. 2017 Apr;47(4):445-454. doi: 10.1097/SHK.0000000000000743.
The molecular basis responsible for tolerance following inflammatory response to lipopolysaccharide (LPS) is not well understood. We hypothesized that inflammation/tolerance in monocytes/ macrophages is dependent on the proteases of proteasome. To test our hypothesis, first, we examined the expression of different proteasome subunits in different human and mouse monocytes/macrophages. Secondly, we investigated the effect of proteasome subunits/ proteases on LPS-induced expression of tumor necrosis factor-α (TNF-α) and nitric oxide (NO) during inflammation and tolerance using mouse RAW 264.7 macrophages, THP1 cells, and cluster of differentiation 14 positive (CD14) human monocytes. We found that RAW 264.7 cells (XYZ), mouse peritoneal resident, thioglycollate-elicited macrophages, primed RAW 264.7 (XYZ, LMP), and human monocytes (LMP) expressed different types of proteasome subunits/activities. Cells containing predominantly either LMP subunits (such as THP-1 and human monocytes), or only X, Y, Z subunits (RAW 264.7 cells not primed) could only induce TNF-α, but not NO, while cells containing all five to six subunits (XYZ, LMP) of the proteasome could induce both mediators in response to LPS. Distinct states of inflammation/tolerance in LPS treated cells, strongly correlated with an upregulation or downregulation of proteasome's subunits (proteases), respectively. Moreover, interferon-γ treatment of tolerant cells caused robust induction of proteasome's subunit expression in mouse macrophages and human monocytes, and cells regained their ability to respond to LPS. These studies are vital for understanding function of proteasome's subunits during inflammation/tolerance in mouse and human cells, and for design of therapeutic strategies for all diseases based on inflammation.
对脂多糖(LPS)炎症反应后耐受性的分子基础尚未完全了解。我们假设单核细胞/巨噬细胞中的炎症/耐受性取决于蛋白酶体的蛋白酶。为了验证我们的假设,首先,我们检测了不同人类和小鼠单核细胞/巨噬细胞中不同蛋白酶体亚基的表达。其次,我们使用小鼠RAW 264.7巨噬细胞、THP1细胞和分化簇14阳性(CD14)人类单核细胞,研究了蛋白酶体亚基/蛋白酶对LPS诱导的炎症和耐受性期间肿瘤坏死因子-α(TNF-α)和一氧化氮(NO)表达的影响。我们发现RAW 264.7细胞(XYZ)、小鼠腹腔驻留的巯基乙酸诱导的巨噬细胞、预处理的RAW 264.7(XYZ,LMP)和人类单核细胞(LMP)表达不同类型的蛋白酶体亚基/活性。主要含有LMP亚基(如THP-1和人类单核细胞)或仅含有X、Y、Z亚基(未预处理的RAW 264.7细胞)的细胞只能诱导TNF-α,而不能诱导NO,而含有蛋白酶体所有五到六个亚基(XYZ,LMP)的细胞对LPS反应可诱导两种介质。LPS处理细胞中不同的炎症/耐受状态,分别与蛋白酶体亚基(蛋白酶)的上调或下调密切相关。此外,用干扰素-γ处理耐受细胞可导致小鼠巨噬细胞和人类单核细胞中蛋白酶体亚基表达的强烈诱导,并且细胞恢复了对LPS的反应能力。这些研究对于理解蛋白酶体亚基在小鼠和人类细胞炎症/耐受性中的功能,以及基于炎症设计所有疾病的治疗策略至关重要。