Chang Sulie L, Huang Wenfei, Mao Xin, Sarkar Sabroni
Institute of NeuroImmune Pharmacology, Seton Hall University, 400 South Orange Avenue, South Orange, NJ 07079, USA.
Department of Biological Sciences, Seton Hall University, 400 South Orange Avenue, South Orange, NJ 07079, USA.
Brain Sci. 2017 Feb 6;7(2):14. doi: 10.3390/brainsci7020014.
Morphine, an effective but addictive analgesic, can profoundly affect the inflammatory response to pathogens, and long-term use can result in morphine tolerance. Inflammasomes are protein complexes involved in the inflammatory response. The nucleotide-binding oligomerization domain-like receptor (NLR) Family Pyrin Domain Containing (NLRP) 12 (NLRP12) inflammasome has been reported to have anti-inflammatory activity. In this study, we examined the expression of NLRP12 inflammasome related genes in the adult F344 rat brain in response to the bacterial endotoxin lipopolysaccharide (LPS) in the presence and absence of morphine tolerance. Morphine tolerance was elicited using the 2 + 4 morphine-pelleting protocol. On Day 1, the rats were pelleted subcutaneously with 2 pellets of morphine (75 mg/pellet) or a placebo; on Days 2 and 4 pellets were given. On Day 5, the animals were randomly assigned to receive either 250 µg/kg LPS or saline (i.p.). The expression of 84 inflammasome related genes in the rat brain was examined using a Ploymerase Chain Reaction (PCR) array. In response to LPS, there was a significant increase in the expression of the pro-inflammatory cytokine/chemokine genes interleukin-1 beta (Il-1β), interleukin-6 (Il-6), C-C motif chemokine ligand 2 (Ccl2), C-C motif chemokine ligand 7 (Ccl7), C-X-C motif chemokine ligand 1 (Cxcl1), and C-X-C motif chemokine ligand 3 (Cxcl3) and a significant decrease in the anti-inflammatory NLRP12 gene in both morphine-tolerant and placebo-control rats compared to saline-treated rats, although the changes were greater in the placebo-control animals. The Library of Integrated Network-Based Cellular Signatures' (LINCS) connectivity map was used to analyze the list of affected genes to identify potential targets associated with the interactions of LPS and morphine tolerance. Our data indicate that, in the morphine tolerant state, the expression of NLRP12 and its related genes is altered in response to LPS and that the Vacuolar protein-sorting-associated protein 28 (VPS28), which is involved in the transport and sorting of proteins into sub-cellular vesicles, may be the key regulator of these alterations.
吗啡是一种有效但会上瘾的镇痛药，可深刻影响对病原体的炎症反应，长期使用会导致吗啡耐受性。炎性小体是参与炎症反应的蛋白质复合物。据报道，含核苷酸结合寡聚化结构域样受体（NLR）家族吡啉结构域（NLRP）12（NLRP12）炎性小体具有抗炎活性。在本研究中，我们检测了成年F344大鼠脑中NLRP12炎性小体相关基因在存在和不存在吗啡耐受性的情况下对细菌内毒素脂多糖（LPS）的反应表达。使用2 + 4吗啡植入方案诱导吗啡耐受性。在第1天，给大鼠皮下植入2粒吗啡（75 mg/粒）或安慰剂；在第2天和第4天给予植入物。在第5天，将动物随机分配接受250 µg/kg LPS或生理盐水（腹腔注射）。使用聚合酶链反应（PCR）阵列检测大鼠脑中84个炎性小体相关基因的表达。与生理盐水处理的大鼠相比，在吗啡耐受和安慰剂对照大鼠中，对LPS的反应导致促炎细胞因子/趋化因子基因白细胞介素-1β（Il-1β）、白细胞介素-6（Il-6）、C-C基序趋化因子配体2（Ccl2）、C-C基序趋化因子配体7（Ccl7）、C-X-C基序趋化因子配体1（Cxcl1）和C-X-C基序趋化因子配体3（Cxcl3）的表达显著增加，抗炎NLRP12基因显著减少，尽管安慰剂对照动物的变化更大。基于综合网络的细胞特征库（LINCS）连接图谱用于分析受影响基因列表，以识别与LPS和吗啡耐受性相互作用相关的潜在靶点。我们的数据表明，在吗啡耐受状态下，NLRP12及其相关基因的表达在对LPS的反应中发生改变，并且参与蛋白质运输和分选到亚细胞小泡中的液泡蛋白分选相关蛋白28（VPS28）可能是这些改变的关键调节因子。
