College of Pharmacy, Gansu University of Traditional Chinese Medicine (TCM), Lanzhou 730000, PR China; Ankang Inspection and Detection Center of Food and Drug Control, Ankang 725000, PR China.
Key Laboratory of New Animal Drug Project of Gansu Province, Key Laboratory of Veterinary Pharmaceutical Development, Ministry of Agriculture, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences, Lanzhou 730050, PR China.
J Pharm Biomed Anal. 2020 Sep 5;188:113374. doi: 10.1016/j.jpba.2020.113374. Epub 2020 May 31.
As a folk medicine, Aconitum sinomontanum Nakai (Ranunculaceae, Gaowutou, in Chinese) is used by traditional healers to treat many disorders, including pain and inflammatory diseases, but it exhibits the toxic side effects. This study aimed to obtain toxic extract parts from A. sinomontanum roots and to further evaluate the antinociceptive and anti-inflammatory effects of toxic extract parts on mice. This work also aimed to identify various chemical compositions of the toxic and active extract parts and evaluate the safety profile of this plant.
Experimental drugs (petroleum ether, chloroform, ethyl acetate, n-butanol, alcohol and water extracts) were obtained through systematic solvent extraction from 95 % ethanol extract from A. sinomontanum roots. An acute toxicity test was conducted to compare the toxicity of different extracts administered at the maximum dose to screen a highly toxic extract. In pharmacodynamic activity analysis, the antinociceptive activity of the A. sinomontanum toxic extract was assessed using an acetic acid-induced abdominal writhing model and a hot plate test. Anti-inflammatory activity was assessed in terms of xylene-induced inflammation. Ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was performed to establish a chromatographic fingerprint and to identify various chemical components of the toxic and active extract.
Chloroform, water and n-butanol extracts elicited significant toxic effects and had LD of 89.65, 1805.40 and 24409.41 mg/kg, respectively. Antinociceptive and anti-inflammatory activities indicated that the chloroform extract significantly alleviated (p < 0.01) the pain induced by acetic acid with an inhibition rate of 44.7 % (5.9 mg/kg) and 50.4 % (17.7 mg/kg). The chloroform extract also significantly (p < 0.01) increased the latency time during the hot plate test. The latency time at 5.9 and 17.7 mg/kg increased from 15.6 ± 4.1 s to 47.3 ± 6.4 s and from 16.3 ± 3.8 s to 49.8 ± 7.6 s (p < 0.01), respectively, 2 h after treatment. In the inflammatory test, the chloroform extract significantly reduced (p < 0.01) the xylene-induced mouse ear oedema with an inhibition rate of 45.48 % (5.9 mg/kg) and 51.46 % (17.7 mg/kg), respectively. This result indicated that A. sinomontanum chloroform extract was also the active extract part of A. sinomontanum. Phytochemical analysis revealed the presence of alkaloids in the chloroform extract. A total of 30 compounds were detected, and 23 compounds, including lappaconine, ranaconidine, 8-O-acetylexcelsine, sinomontanine H, finaconitine, lappacontine, N-dacetyllappaconitine, ranaconitine and isolappaconitine, were identified.
A. sinomontanum chloroform extract possesses antinociceptive and anti-inflammatory activities and exhibits significant toxic effects. Phytochemical analysis indicated that some alkaloids may be the main bioactive ingredient responsible for the toxicity and efficacy of A. sinomontanum. This work contributes to the determination of the safety of the medicinal use of A. sinomontanum roots.
作为一种民间药物，高山乌头（毛茛科，高头，中文）被传统医生用于治疗多种疾病，包括疼痛和炎症性疾病，但它表现出毒性副作用。本研究旨在从高山乌头的根部分离出有毒提取物部分，并进一步评估有毒提取物部分对小鼠的镇痛和抗炎作用。本工作还旨在鉴定有毒和有效提取物部分的各种化学成分，并评估该植物的安全性概况。
通过系统溶剂从 95%乙醇提取的高山乌头根部分离得到实验药物（石油醚、氯仿、乙酸乙酯、正丁醇、醇和水提取物）。急性毒性试验比较了不同提取物在最大剂量下的毒性，以筛选高毒性提取物。在药效学分析中，采用醋酸诱导的腹部扭体模型和热板试验评估高山乌头有毒提取物的镇痛活性。采用二甲苯诱导的炎症评估抗炎活性。采用超高效液相色谱-四极杆飞行时间质谱（UPLC-Q/TOF-MS）建立色谱指纹图谱，并鉴定有毒和有效提取物的各种化学成分。
氯仿、水和正丁醇提取物表现出显著的毒性作用，LD 分别为 89.65、1805.40 和 24409.41mg/kg。镇痛和抗炎活性表明，氯仿提取物显著缓解（p<0.01）醋酸引起的疼痛，抑制率分别为 44.7%（5.9mg/kg）和 50.4%（17.7mg/kg）。氯仿提取物还显著（p<0.01）增加了热板试验中的潜伏期。5.9 和 17.7mg/kg 处理后 2 小时，潜伏期从 15.6±4.1s 分别增加到 47.3±6.4s 和 49.8±7.6s（p<0.01）。在炎症试验中，氯仿提取物显著抑制（p<0.01）二甲苯诱导的小鼠耳肿胀，抑制率分别为 45.48%（5.9mg/kg）和 51.46%（17.7mg/kg）。这表明高山乌头氯仿提取物也是高山乌头的有效提取物部分。植物化学分析表明，氯仿提取物中存在生物碱。共检测到 30 种化合物，其中 23 种化合物，包括拉帕科宁、拉那可丁、8-O-乙酰高丙氨酸、山乌头碱 H、非那定碱、拉帕可宁、N-乙酰拉帕可宁、拉那可丁和异拉帕可宁被鉴定出来。
高山乌头氯仿提取物具有镇痛和抗炎作用，并表现出显著的毒性作用。植物化学分析表明，一些生物碱可能是高山乌头毒性和功效的主要生物活性成分。这项工作有助于确定高山乌头根的药用安全性。
