Neoline, fuziline, songorine and 10-OH mesaconitine are potential quality markers of Fuzi: In vitro and in vivo explorations as well as pharmacokinetics, efficacy and toxicity evaluations
Neoline, fuziline, songorine and 10-OH mesaconitine are potential quality markers of Fuzi: In vitro and in vivo explorations as well as pharmacokinetics, efficacy and toxicity evaluations
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Neoline、fuziline、songorine 和 10-OH mesaconitine 是附子的潜在质量标志物：体外和体内探索以及药代动力学、功效和毒性评价
DOI：
10.1016/j.jep.2022.115879
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发表时间：
2022-11-28
影响因子：
5.4
通讯作者：
Zhu，Lijun
中科院分区：
文献类型：
--
作者：
Li，Xiaocui;Hou，Weiqing;Zhu，Lijun
附子是附子的侧根，在治疗阳虚寒凝证方面具有不可替代的作用。然而，附子的安全边际很小，因为它的药理成分乌头生物碱具有潜在的心脏毒性和神经毒性。目前控制附子药效和毒性的质量标记物(Q标记物)是3个单酯二萜生物碱，即苯甲酰乌头碱(BAC)、苯甲酰次乌头碱(BMA)和3个二酯二萜生物碱(AC)、次乌头碱(MA)和新乌头碱(MA)。然而，越来越多的证据表明，目前的6个Q标记对附子来说可能还不够有效或毒性特异性。本研究的目的是探索和评价附子的有效性或毒性特异性潜在质量标记(PQ标记)。材料和方法通过对30个小鼠暴露的药材和生物碱进行分析，探索PQ标记。测定了PQ标志物在C57BL/6J小鼠体内的药代动力学。采用λ-角叉菜胶足肿胀模型和脂多糖诱导的RAW264.7细胞炎症模型评价PQ标记物的抗炎作用，采用醋酸致痛模型和Hargreaves试验评价PQ标记物的镇痛作用。结果经体外和体内实验，共发现7种PQ标记物，分别为Neoline(NE)、Fuziline(FE)、Songorine(SE)、10-OH新乌头碱(10-OHMA)、他拉西明、异他拉替丁和16β-OH。在中草药中，NE、FE、SE和10-OHMA的含量高于其他生物碱。附子样品中NE、FE和SE的含量均远高于BAC，且有56.67%的样品中10-羟基-MA含量高于AC。在小鼠血浆和组织中，NE、FE、SE、他拉西明、异塔拉西定和16β-OH的含量高于其他生物碱，包括6种当前的Q标记。对NE、FE、SE和10-OHMA的药代动力学、疗效和毒性进行了进一步评价。NE(63.82%)、FE(18.14%)和SE(49.51%)的平均生物利用度高于BMA(3.05%)。NE、FE和SE具有剂量依赖性的抗炎和镇痛作用，且作用强于BMA。同时，NE、FE和SE对小鼠的心脏毒性和神经毒性均低于BMA，而BMA可显著提高小鼠的肌酸激酶活性和基质金属蛋白酶9水平。10-OH-MA的平均口服生物利用度(7.02%)高于MA(1.88%)。小鼠静脉注射10-羟基-MA的半数致死量(LD50)为0.11 mg/kg，与MA(0.13 mg/kg)相近。10-OHMA具有明显的心脏毒性和神经毒性，显著的抗炎、镇痛作用，与MA相当。结论附子在体外和体内实验中发现7个PQ标志物。其中，NE、FE和SE的药效特异性强于BMA，10-羟基MA的毒性特异性与MA相同。
Ethnopharmacological relevanceFuzi, the lateral roots of Aconitum carmichaelii Debx, plays an irreplaceable role in treatingYangdeficiency and cold coagulation syndromes. However, Fuzi has a narrow margin of safety since its pharmacological constituents,Aconitumalkaloids, have potential cardiotoxicity and neurotoxicity. The current quality markers (Q-markers) for the control of Fuzi's efficacy and toxicity are 3 monoester-diterpenoid alkaloids, namely, benzoylaconine (BAC), benzoylhypaconine and benzoylmesaconine (BMA) and 3 diester-diterpenoid alkaloids, namely, aconitine (AC), hypaconitine and mesaconitine (MA). However, mounting evidence indicates that the current 6 Q-markers may not be efficacy- or toxicity-specific enough for Fuzi.Aim of the studyThe aim of this study was to explore and evaluate efficacy- or toxicity-specific potential quality markers (PQ-markers) of Fuzi.Materials and methodsPQ-markers were explored by analyzing 30 medicinal samples and alkaloids exposed in mouse. Pharmacokinetics of PQ-markers on C57BL/6J mice were determined. Anti-inflammatory effects of PQ-markers were evaluated by λ-carrageenan-induced paw edema model and lipopolysaccharide-induced RAW264.7 cell inflammatory model, while analgesic effects were assessed by acetic acid-induced pain model and Hargreaves test. Cardiotoxicity and neurotoxicity of PQ-markers were assessed by histological and biochemical analyses, while acute toxicity was evaluated by modified Kirschner method.ResultsAfterin vitroandin vivoexplorations, 7 PQ-markers, namely, neoline (NE), fuziline (FE), songorine (SE), 10-OH mesaconitine (10-OH MA), talatizamine, isotalatizidine and 16β-OH cardiopetalline, were found. In the herbal medicines, NE, FE, SE and 10-OH MA were found in greater abundance than many other alkaloids. Specifically, the amounts of NE, FE and SE in the Fuzi samples were all far higher than that of BAC, and the contents of 10-OH MA in 56.67% of the samples were higher than that of AC. In mouse plasma and tissues, NE, FE, SE, talatizamine, isotalatizidine and 16β-OH cardiopetalline had higher contents than the other alkaloids, including the 6 current Q-markers. The pharmacokinetics, efficacy and toxicity of NE, FE, SE and 10-OH MA were further evaluated. The average oral bioavailabilities of NE (63.82%), FE (18.14%) and SE (49.51%) were higher than that of BMA (3.05%). Additionally, NE, FE and SE produced dose-dependent anti-inflammatory and analgesic effects, and their actions were greater than those of BMA. Concurrently, the toxicities of NE, FE and SE were lower than those of BMA, since no cardiotoxicity or neurotoxicity was found in mice after NE, FE and SE treatment, while BMA treatment notably increased the creatine kinase activity and matrix metalloproteinase 9 level in mice. The average oral bioavailability of 10-OH MA (7.02%) was higher than that of MA (1.88%). The median lethal dose (LD50) of 10-OH MA in mice (0.11 mg/kg) after intravenous injection was close to that of MA (0.13 mg/kg). Moreover, 10-OH MA produced significant cardiotoxicity and neurotoxicity, and notable anti-inflammatory and analgesic effects that were comparable to those of MA.ConclusionsSeven PQ-markers of Fuzi were found afterin vitroandin vivoexplorations. Among them, NE, FE and SE were found to be more efficacy-specific than BMA, and 10-OH MA was as toxicity-specific as MA.