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神经病理性疼痛(NPP)的新型治疗药物以及治疗靶点亟待研究。中国钩吻为马钱科植物胡蔓藤的全草,盛产于闽、粤、桂等地。钩吻的主要有效成分为吲哚类生物碱,含有数十种生物碱单体,其中钩吻素子(koumine,KM)为国产钩吻生物碱单体中含量最高的一种有效成分。民间一直应用钩吻原植物治疗各类疼痛,尤其神经性疼痛与癌性疼痛。有报道,钩吻总碱具有显著的镇痛效果,但治疗指数小,有效量与中毒量较接近。本文拟从国产钩吻分离纯化获取钩吻素子,并报道其抗神经病理性疼痛作用及其机制。选取本地采集的钩吻根和茎,提取钩吻总生物碱;应用高速逆流色谱等技术分离纯化钩吻总碱获得钩吻素子单体,纯度达99%;经紫外光谱、红外光谱、核磁共振谱、质谱分析、元素分析以及熔点测定等确证其化学结构,钩吻素子分子式为C20H22N2O,分子量306.3。钩吻素子原料药对光、温度、湿度稳定,钩吻素子水溶液在pH值3～8以及温度在室温、40℃、60℃条件下稳定。在大小鼠坐骨神经慢性束缚损伤(CCI)模型和脊神经结扎模型上,以动物机械阈值、热阈值和自发痛行为为指标,首次观察到ig或sc钩吻素子具有显著的抗NPP作用,有效剂量远低于中毒剂量(小鼠sc钩吻素子LD50为99 mg·kg-1)。脊髓可能是钩吻素子抗NPP的重要作用部位;脊髓神经活性甾体3α,5α-四氢孕酮(3α,5α-THP)参与调控NPP,钩吻素子可显著提高CCI模型大鼠脊髓腰段内3α,5α-THP的水平;3α-羟化类固醇脱氢酶(3α-HSD)是催化3α,5α-THP合成的关键酶,钩吻素子可显著上调脊髓3α-HSD酶的表达。上述结果表明,钩吻素子具有显著的抗NPP作用;增强脊髓3α-HSD酶的表达,促进脊髓3α,5α-THP的生成,可能是其发挥抗NPP作用的机制之一。
New types of neuropathic pain (NPP) therapeutic drugs and therapeutic targets need to be studied. China hooked kiss for the whole plant Sika money horse plants, rich in Fujian, Guangdong, Guangxi and other places. The main active ingredient of hook kiss is indole alkaloid, which contains dozens of alkaloid monomers, of which koumine (KM) is the highest active ingredient in domestic kouhee alkaloids. The folk have been using the kiss kiss original plant treatment of various types of pain, especially neuropathic pain and cancer pain. It has been reported that hook kisses alkaloids have a significant analgesic effect, but the therapeutic index is small, the effective amount of poisoning and more close. In this paper, we will purify and obtain koukin from Chinese hook kisses and report its anti-neuropathic pain and its mechanism. The root and stem of Gynostemma pentaphyllum collected locally were extracted, and the total alkaloids of Gynostemma pentaphylla were extracted. High-speed countercurrent chromatography and other techniques were used to separate and purify gouache kinin. The purity was 99% .Ultraviolet spectroscopy, infrared spectroscopy, The chemical structure was confirmed by spectroscopy, mass spectrometry, elemental analysis and melting point determination. The molecular formula of CnB was C20H22N2O and the molecular weight was 306.3. Hook kokin raw material medicine on the light, temperature, humidity and stability, kiss koi water solution at pH 3 to 8 and the temperature at room temperature, 40 ℃, 60 ℃ under the conditions of stability. In the chronic constriction injury (CCI) model and spinal nerve ligation model of sciatic nerve in rats and rats, we found that ig or sc-kinkin has a significant anti-NPP effect with animal’s mechanical threshold, thermal threshold and spontaneous pain behavior as the effective dose Lower than the poisoning dose (mice c-kiss kinin LD50 99 mg · kg-1). Spinal cord may be an important site of anti-NPP of cynomolgus kina.Nerpine-active steroid 3α, 5α-tetrahydrogesterone (3α, 5α-THP) is involved in the regulation of NPP and koumissin can significantly improve the spinal cord lumbar 3α, 5α-THP. 3α-hydroxysteroid dehydrogenase (3α-HSD) is the key enzyme in the synthesis of 3α, 5α-THP. Goukonin can significantly up-regulate the expression of 3α-HSD enzyme in the spinal cord. The above results show that koumin has a significant anti-NPP effect. Increasing the expression of 3α-HSD enzyme in the spinal cord and promoting the production of 3α, 5α-THP in the spinal cord may be one of the mechanisms by which it exerts its anti-NPP effect.