Studies in~a new pain control by spinorphin, an endogenous factor : Changes in level of spinorphin and dipeptidyl peptidase III in cerebrospinal fluid from patients with pain

脊髓啡肽（LVVYPWT）是一种从脊髓中纯化的内源性生物活性肽，在大鼠缓激肽诱导的疼痛中显示出伤害性作用，在小鼠急性炎症模型中显示出抑制卡拉胶诱导的中性粒细胞积累的活性。为了进一步明确其在疼痛和炎症中的生理作用，我们建立了一种定量测定人脑脊液（CSE）中spinorphin的方法，并分析了其水平与生理状态的关系。首先，用抗兔spinorphin抗体建立了特异性、敏感性的竞争酶免疫分析法（EIA）。特异性抗体与spinorphin类似物的交叉反应性不显著，表明EIA对spinorphin具有特异性。其次，在除癌性疼痛以外的各种类型疼痛患者的csf中，疼痛组的spinorphin水平高于对照组（无疼痛和非炎症性中枢神经系统疾病患者）（21.4±8.2 ng/ml, n=48 vs 8.61±4.4 ng/ml, n=28，P<0.05）。此外，研究人员还检测了几种调节某些类型疼痛的脊髓啡加工酶的活性。疼痛组csf中二肽基肽酶III （DPPIII）活性低于对照组（1.40±0.65 vs 2.05±0.88 pmol/30 min/100μ1，P<0.05）。有趣的是，spinorphin水平升高与DPPIII水平降低之间存在统计学相关性（r=-0.514，P<0.05）。这些结果表明，spinorphin的水平可能通过酶的调节作用，作为病理生理状态的一个指标。

Spinorphin (LVVYPWT), an endogenous bioactive peptide purified from the spinal cord, demonstrates a nociceptive effect on bradykinin-induced pain in the rat and inhibitory activity against carrageenan-induced neutrophil accumulation in the mouse model of acute inflammation. To further clarify its physiological role in pain and inflammation, we established a method to quantify spinorphin in human cerebrospinal fluid (CSE) and analyzed the relation between its level and the physiological state. First, a specific and sensitive competitive enzyme immunoassay (EIA) was developed with an anti-rabbit spinorphin antibody. Cross-reactivity of the specific antibody with spinorphin analogues was not significant, suggesting that the EIA was specific to spinorphin. Next, levels of spinorphin measured in CSFs derived from patients with various types of pain other than cancer pain were higher in the pain group than in the controls (patients with pain-free and noninflammatory disease of the central nervous system) (21.4±8.2 ng/ml, n=48 vs 8.61±4.4 ng/ml, n=28,P<0.05). Furthermore, the activity of several spinorphin-processing enzymes that regulate some types of pain was examined. The activity of dipeptidyl peptidase III (DPPIII) in CSFs of the pain group was lower than that in the control (1.40±0.65 vs 2.05±0.88 pmol/30 min/100μ1,P<0.05). Interestingly, a statistical correlation was observed between the increase of spinorphin levels and the decrease of DPPIII (r=-0.514,P<0.05). These results indicate that the level of spinorphin may, through the regulatory effect of enzymes, be an indicator of the pathophysiological state.