Mićović Tijana, Katanić Stanković Jelena S, Bauer Rudolf, Nöst Xuehong, Marković Zoran, Milenković Dejan, Jakovljević Vladimir, Tomović Marina, Bradić Jovana, Stešević Danijela, Stojanović Danilo, Maksimović Zoran
Institute for Medicines and Medical Devices of Montenegro, Bulevar Ivana Crnojevića 64a, 81000, Podgorica, Montenegro.
Institute for Information Technologies Kragujevac, Department of Science, University of Kragujevac, Jovana Cvijića bb, 34000, Kragujevac, Serbia.
J Ethnopharmacol. 2022 Jul 15;293:115201. doi: 10.1016/j.jep.2022.115201. Epub 2022 Mar 28.
Medicinal properties of hyssop have been used in traditional medicine since ancient times, inter alia, in diseases/conditions with an inherent inflammatory process.
Accordingly, the aim of this study was to investigate the anti-inflammatory properties of hyssop herb preparations (essential oil and methanol extracts) in vivo, in vitro and in silico.
For in vitro testing of essential oils and extracts of hyssop herb, the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzyme assays were used. In vivo anti-inflammatory potential of the extracts (at doses of 50, 100 and 200 mg/kg) was assessed using the carrageenan-induced rat paw edema test. Molecular docking and dynamics were used for in silico testing of the inhibitory activity of chlorogenic (CA) and rosmarinic (RA) acids, as the dominant compounds in the tested methanol extracts against COX-1 and COX-2 enzymes.
Significant inhibitory activity was shown in the COX-2 test regarding extracts (essential oils did not exhibit any significant activity). Namely, all analyzed extracts, at a concentration of 20 μg/mL, showed a percentage of inhibition of COX-2 enzyme (54.04-63.04%), which did not indicate a statistically significant difference from the positive control of celecoxib (61.60%) at a concentration of 8.8 μM. In vivo testing showed that all methanol extracts of hyssop herb, at the highest test dose of 200 mg/kg in the third and fourth hours, after carrageenan administration, exhibited a statistically significant (p < 0.05) inhibitory effect on the increase in rat paw edema in relation to control. This activity is comparable or higher in relation to the reference substance, indomethacin, at a concentration of 8 mg/kg. The preliminary in silico results suggest that investigated compounds (RA and CA) showed better inhibitory activity against COX-1 and COX-2 than standard non-steroidal anti-inflammatory drug (NSAID), ibuprofen, as evident from the free binding energy (ΔG in kJ mol). The binding energies of the docked compounds to COX-1 and -2 were found to be in the range between -47.4 and -49.2 kJ mol. Ibuprofen, as the one NSAID, for the same receptors targets, showed remarkably higher binding energy (ΔG = -31.3 kJ mol to COX-1, and ΔG = -30.9 kJ mol to COX-2).
The results obtained not only support the traditional use of hyssop herb in inflammatory conditions in folk medicine, but also open the door to and the need for further in vivo testing of extracts in order to examine the molecular mechanism of anti-inflammatory activity in living systems and possibly develop a new anti-inflammatory drug or supplement.
自古以来，牛膝草的药用特性就在传统医学中有所应用，尤其用于治疗具有内在炎症过程的疾病/病症。
因此，本研究的目的是在体内、体外和计算机模拟条件下研究牛膝草制剂（精油和甲醇提取物）的抗炎特性。
为了对牛膝草的精油和提取物进行体外测试，采用了环氧合酶-1（COX-1）和环氧合酶-2（COX-2）酶检测法。使用角叉菜胶诱导的大鼠足趾肿胀试验评估提取物（剂量为50、100和200mg/kg）的体内抗炎潜力。分子对接和动力学用于对绿原酸（CA）和迷迭香酸（RA）的抑制活性进行计算机模拟测试，这两种酸是测试甲醇提取物中针对COX-1和COX-2酶的主要化合物。
在COX-2检测中，提取物显示出显著的抑制活性（精油未表现出任何显著活性）。具体而言，所有分析的提取物在浓度为20μg/mL时，对COX-2酶的抑制率为54.04%-63.04%，与浓度为8.8μM的塞来昔布阳性对照（61.60%）相比，差异无统计学意义。体内测试表明，牛膝草的所有甲醇提取物在角叉菜胶给药后的第三和第四小时，以最高测试剂量200mg/kg时，与对照组相比，对大鼠足趾肿胀的增加表现出统计学显著（p<0.05）的抑制作用。与浓度为8mg/kg的参比物质吲哚美辛相比，该活性相当或更高。初步的计算机模拟结果表明，所研究的化合物（RA和CA）对COX-1和COX-2的抑制活性优于标准非甾体抗炎药布洛芬，从自由结合能（ΔG，单位为kJ/mol）可以明显看出。对接化合物与COX-1和-2的结合能在-47.4至-49.2kJ/mol之间。作为一种非甾体抗炎药，布洛芬针对相同的受体靶点，显示出明显更高的结合能（对COX-1为ΔG=-31.3kJ/mol，对COX-2为ΔG=-30.9kJ/mol）。
所获得的结果不仅支持了民间医学中牛膝草在炎症病症中的传统应用，也为进一步进行提取物的体内测试打开了大门并提出了需求，以便研究其在生物系统中的抗炎活性分子机制，并有可能开发出一种新的抗炎药物或补充剂。
