Zhang Jing-Xian, Li Shang-Rong, Yao Shuai, Bi Qi-Rui, Hou Jin-Jun, Cai Lu-Ying, Han Su-Mei, Wu Wan-Ying, Guo De-An
National Engineering Laboratory for TCM Standardization Technology, Shanghai Research Center for Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, China.
National Engineering Laboratory for TCM Standardization Technology, Shanghai Research Center for Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, China.
J Ethnopharmacol. 2016 Apr 2;181:229-35. doi: 10.1016/j.jep.2016.01.039. Epub 2016 Jan 27.
Pearl and nacre are valuable traditional medicines to treat palpitations, convulsions or epilepsy in China for thousands of years. However, the active ingredients are not clear till now.
The main purpose of the current investigation was to assess the anticonvulsant and sedative-hypnotic activity of pearl powder and nacre powder, including their corresponding 6 protein extracts.
Determination of the amino acid composition of the obtained protein was carried out by ultra-performance liquid chromatography (UPLC) combined with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) pre-column derivatisation. The influence of the tested drugs on locomotor activity and convulsions latency was recorded. The contents of 5-Hydroxytryptamine (5-HT) and γ-aminobutyric acid (GABA) in brain were detected by enzyme-linked immunesorbent assay (ELISA) kits. In addition, immunohistochemistry was carried out to evaluate the changes of 5-HT3 and GABAB. In parallel, the expressions of them were demonstrated by western blot.
The obtained data suggested that pearl original powder (1.1g/kg), pearl water-soluble protein (0.2g/kg), pearl acid-soluble protein (0.275g/kg), pearl conchiolin protein (1.1g/kg), nacre original powder (1.1g/kg), nacre water-soluble protein (0.2g/kg), nacre acid-soluble protein (0.7g/kg) and nacre conchiolin protein (1.1g/kg) could down-regulate the expression of 5-HT3 and up-regulate the level of GABAB to varying degrees compared with the control group. Besides, drug administration also reduced the locomotor activity and increased convulsions latency with a certain mortality.
These findings correlated with the traditional use of pearl and nacre as sedation and tranquilization agents, thus making them interesting sources for further drug development and also providing critical important evidence for the selection of quality control markers.
在中国,珍珠和珍珠母作为治疗心悸、惊厥或癫痫的珍贵传统药物已有数千年历史。然而,其活性成分至今仍不明确。
本研究的主要目的是评估珍珠粉和珍珠母粉及其相应的6种蛋白质提取物的抗惊厥和镇静催眠活性。
采用超高效液相色谱(UPLC)结合6-氨基喹啉-N-羟基琥珀酰亚胺基氨基甲酸酯(AQC)柱前衍生化法测定所得蛋白质的氨基酸组成。记录受试药物对运动活性和惊厥潜伏期的影响。采用酶联免疫吸附测定(ELISA)试剂盒检测脑中5-羟色胺(5-HT)和γ-氨基丁酸(GABA)的含量。此外,进行免疫组织化学以评估5-HT3和GABAB的变化。同时,通过蛋白质印迹法检测它们的表达。
所得数据表明,与对照组相比,珍珠原粉(1.1g/kg)、珍珠水溶性蛋白(0.2g/kg)、珍珠酸溶性蛋白(0.275g/kg)、珍珠壳硬蛋白(1.1g/kg)、珍珠母原粉(1.1g/kg)、珍珠母水溶性蛋白(0.2g/kg)、珍珠母酸溶性蛋白(0.7g/kg)和珍珠母壳硬蛋白(1.1g/kg)均可不同程度地下调5-HT3的表达并上调GABAB的水平。此外,给药还降低了运动活性,延长了惊厥潜伏期,并有一定的死亡率。
这些发现与珍珠和珍珠母作为镇静安神药物的传统用途相关,因此使其成为进一步药物开发的有趣来源,也为质量控制标志物的选择提供了至关重要的证据。