Kim Chung Su, Ko Justin Sangwook, Lee Ae Ryoung, Shin Byung Seop, Choi Soo Joo, Lee Jeong Jin, Kim Hyun Soo, Lee Sangmin Maria
Department of Anesthesiology and Pain Medicine, Samsung Medical Center, College of Medicine, Sungkyunkwan University, Seoul, Korea.
Curr Ther Res Clin Exp. 2011 Feb;72(1):23-35. doi: 10.1016/j.curtheres.2011.02.004.
The effect of opioids on inflammation and immune responses is an important subject of investigation because immunoregulatory cytokines are produced in the central nervous system and opioid receptors are widespread in these cells.
The aim of this study was to evaluate the immunomodulatory effect of morphine on the C3 expression (both constitutive and proinflammatory cytokine-induced C3 expression) in primary rat astrocytes.
Primary rat astrocytes were untreated or treated with morphine in different concentrations (10(-6) to 10(-2) M) before incubation without or with 5 U/mL tumor necrosis factor-α (TNF-α), and C3 protein and mRNA expressions were measured. Similarly, astrocytes were treated with 10(-3) M morphine and stimulated with other proinflammatory cytokines, including 10 ng/mL interleukin-8 (IL-8) and 5 U/mL IL-1β. Astrocytes were exposed to 10(-5) M naloxone for 2 hours before adding morphine, and TNF-α and C3 protein was measured. Tumor growth factor-β (TGF-β) was measured from the supernatants of each proinflammatory cytokine.
All results are expressed as mean percentages of C3 production by normalizing C3 without morphine or any cytokine treatment as 100%. Constitutive C3 protein production was decreased at morphine 10(-3) M (57.2%) and 10(-2) M (30.1%). Pretreatment with morphine suppressed induction of C3 expression at both the protein and mRNA levels in astrocytes stimulated with TNF-α, IL-8, and IL-1β (P < 0.05) in a dose-dependent manner. The inhibition of C3 protein production by morphine (10(-3) M; 33%) was partially attenuated by naloxone (52.0%) (P < 0.05). The pretreatment of astrocytes with morphine (10(-3) M) before stimulation with TNF-α, IL-8, and IL-1β increased by 33% (P < 0.05), decreased by 15.2% (P < 0.05), and did not change the production of TGF-β protein, respectively.
Morphine downregulated both constitutive and proinflammatory cytokine-induced C3 expression of astrocytes at the transcriptional level, but not in a cytokine-specific manner.
阿片类药物对炎症和免疫反应的影响是一个重要的研究课题，因为免疫调节细胞因子在中枢神经系统中产生，且阿片受体广泛存在于这些细胞中。
本研究旨在评估吗啡对原代大鼠星形胶质细胞中C3表达（组成型和促炎细胞因子诱导的C3表达）的免疫调节作用。
原代大鼠星形胶质细胞在不添加或添加5 U/mL肿瘤坏死因子-α（TNF-α）孵育前，不处理或用不同浓度（10⁻⁶至10⁻² M）的吗啡处理，然后检测C3蛋白和mRNA表达。同样，星形胶质细胞用10⁻³ M吗啡处理并用其他促炎细胞因子刺激，包括10 ng/mL白细胞介素-8（IL-8）和5 U/mL白细胞介素-1β（IL-1β）。在添加吗啡前，星形胶质细胞暴露于10⁻⁵ M纳洛酮2小时，然后检测TNF-α和C3蛋白。从每种促炎细胞因子的上清液中检测肿瘤生长因子-β（TGF-β）。
所有结果均以将未用吗啡或任何细胞因子处理时的C3产生量作为100%进行归一化后的C3产生量的平均百分比表示。在吗啡浓度为10⁻³ M（57.2%）和10⁻² M（30.1%）时，组成型C3蛋白产生量降低。用吗啡预处理可抑制TNF-α、IL-8和IL-1β刺激的星形胶质细胞中C3在蛋白和mRNA水平的表达诱导（P<0.05），且呈剂量依赖性。吗啡（10⁻³ M；33%）对C3蛋白产生的抑制作用被纳洛酮部分减弱（52.0%）（P<0.05）。在用TNF-α、IL-8和IL-1β刺激前，用吗啡（10⁻³ M）预处理星形胶质细胞，TGF-β蛋白产生量分别增加33%（P<0.05）、降低15.2%（P<0.05）且无变化。
吗啡在转录水平下调星形胶质细胞组成型和促炎细胞因子诱导的C3表达，但并非以细胞因子特异性方式下调。
