白细胞介素-1β诱导未处理和吗啡脱敏的 U87 MG 人星形细胞瘤细胞中阿片受体的上调。
Interleukin-1 beta-induced up-regulation of opioid receptors in the untreated and morphine-desensitized U87 MG human astrocytoma cells.
机构信息
Institute of NeuroImmune Pharmacology, Seton Hall University, 400 South Orange Ave, South Orange, NJ 07079, USA.
出版信息
J Neuroinflammation. 2012 Nov 20;9:252. doi: 10.1186/1742-2094-9-252.
BACKGROUND
Interleukin-1beta (IL-1β) is a pro-inflammatory cytokine that can be produced in the central nervous system during inflammatory conditions. We have previously shown that IL-1β expression is altered in the rat brain during a morphine tolerant state, indicating that this cytokine may serve as a convergent point between the immune challenge and opiate mediated biological pathways. We hypothesized that IL-1β up-regulates opioid receptors in human astrocytes in both untreated and morphine-desensitized states.
METHODS
To test this hypothesis, we compared the basal expression of the mu (MOR), delta (DOR), and kappa (KOR) opioid receptors in the human U87 MG astrocytic cell line to SH-SY5Y neuronal and HL-60 immune cells using absolute quantitative real time RT-PCR (AQ-rt-RT-PCR). To demonstrate that IL-1β induced up-regulation of the MOR, DOR and KOR, U87 MG cells (2 x 105 cells/well) were treated with IL-1β (20 ng/mL or 40 ng/mL), followed by co-treatment with interleukin-1 receptor antagonist protein (IL-1RAP) (400 ng/mL or 400 ng/mL). The above experiment was repeated in the cells desensitized with morphine, where U87 MG cells were pre-treated with 100 nM morphine. The functionality of the MOR in U87 MG cells was then demonstrated using morphine inhibition of forksolin-induced intracellular cAMP, as determined by radioimmunoassay.
RESULTS
U87 MG cells treated with IL-1β for 12 h showed a significant up-regulation of MOR and KOR. DOR expression was also elevated, although not significantly. Treatment with IL-1β also showed a significant up-regulation of the MOR in U87 MG cells desensitized with morphine. Co-treatment with IL-1β and interleukin-1 receptor antagonist protein (IL-1RAP) resulted in a significant decrease in IL-1β-mediated MOR up-regulation.
CONCLUSION
Our results indicate that the pro-inflammatory cytokine, IL-1β, affects opiate-dependent pathways by up-regulating the expression of the MOR in both untreated and morphine-desensitized U87 MG.
背景
白细胞介素-1β（IL-1β）是一种促炎细胞因子，可在炎症状态下在中枢神经系统中产生。我们之前已经表明，在吗啡耐受状态下，大鼠大脑中的 IL-1β表达发生改变，这表明该细胞因子可能是免疫挑战和阿片类药物介导的生物学途径之间的汇聚点。我们假设 IL-1β在未经处理和吗啡脱敏状态下上调人星形胶质细胞中的阿片受体。
方法
为了验证这一假设，我们使用绝对定量实时 RT-PCR（AQ-rt-RT-PCR）比较了人 U87 MG 星形胶质细胞系中 μ（MOR）、δ（DOR）和 κ（KOR）阿片受体的基础表达与 SH-SY5Y 神经元和 HL-60 免疫细胞。为了证明 IL-1β诱导 MOR、DOR 和 KOR 的上调，用 IL-1β（20ng/mL 或 40ng/mL）处理 U87 MG 细胞（2×105 个细胞/孔），然后用白细胞介素-1 受体拮抗剂蛋白（IL-1RAP）（400ng/mL 或 400ng/mL）共同处理。在用吗啡脱敏的细胞中重复了上述实验，其中 U87 MG 细胞用 100nM 吗啡预处理。然后使用吗啡抑制叉头框蛋白诱导的细胞内 cAMP 来证明 U87 MG 细胞中 MOR 的功能，通过放射免疫测定法确定。
结果
用 IL-1β 处理 12 小时的 U87 MG 细胞显示 MOR 和 KOR 的表达显著上调。DOR 的表达也升高，尽管不显著。用 IL-1β 处理还显示吗啡脱敏的 U87 MG 细胞中 MOR 的表达显著上调。IL-1β 和白细胞介素-1 受体拮抗剂蛋白（IL-1RAP）的共同处理导致 IL-1β 介导的 MOR 上调显著降低。
结论
我们的结果表明，促炎细胞因子 IL-1β 通过上调未处理和吗啡脱敏的 U87 MG 中 MOR 的表达来影响阿片类药物依赖途径。
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