Microstructure, Content and in vitro Release of Brucine and Strychnine in Strychnos Nux-Vomica Powder with Different Particle Sizes

To explore the effect of particle size on the quality uniformity and in vitro release performance of Strychnos nux-vomica powder, seven samples of Strychnos nux-vomica powder with different particle sizes were prepared.Microstructures and particle sizes were analyzed, and high performance liquid chromatography(HPLC) was used to test the contents and in vitro release performances of brucine and strychnine in the samples. Results showed that the contents and the in vitro release rates of brucine(or strychnine) in different samples were different since there are different proportions of endosperms to epidermal cells in Strychnos nux-vomica powder with different particle sizes. Brucine and strychnine in each sample were promptly released in the first ten minutes and their cumulative release rates were higher than 70% after ten minutes. Eighty minutes later, the cumulative release rate tended to be a constant. Considering the quality uniformity and safety of Strychnos nux-vomica powder used as traditional Chinese medicine, it would be better to control the particle size of Strychnos nux-vomica powder between 100 and 140 mesh in which the maximum cumulative release rate in vitro of brucine and strychnine can be relatively low within this range.

The mechanism of hepatotoxicity of Nux Vomica:a network-pharmacology-based study

Objective:To explore the potential mechanism of hepatotoxicity induced by Nux Vomica through network toxicology.Methods:The active components and targets of Nux Vomica were identified and screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform Database(TCMSP),literature research,PubChem Database,Swiss Target Prediction database,etc.Genecards,pharmGKB and OMIM databases were used to collect hepatotoxicity related targets,then,cross them with active component targets to obtain potential targets of hepatotoxicity caused by Nux Vomica.A"Nux Vomica-Potential active components-Potential targets-Hepatotoxicity"network was constructed with Cytoscape 3.8.0 software.The String 11.0 database was used to construct the protein-protein interaction(PPI)network of the targets and to screen out the core targets.In addition,Gene Ontology(GO)function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were conducted by R software,and then the obtained pathways directly related to hepatotoxicity were integrated.Results:In this study,37 active components were screened via TCMSP and literature research,468 targets for the active components of Nux Vomica were obtained.There were 533 hepatotoxicity-related targets,73 potential targets for hepatotoxicity caused by Nux Vomica,and 26 potential active components,among which Ferulic acid,Novacine,Icajine,Simiarenol were the key active components for hepatotoxicity caused by Nux Vomica,and JUN,RELA,and STAT3 were the core target proteins of hepatotoxicity caused by Nux Vomica.There were 1859 GO entries(P-value<0.05),including 1709 entries of Biological Process(BP),39 entries of Cellular Component(CC),and 111 entries of Molecular Function(MF).KEGG enrichment analysis revealed 145 pathways(value<0.05),of which PI3K/AKT signaling pathway,HIF-1 signaling pathway,EGFR tyrosine kinase inhibitor resistance were strongly correlated with the hepatotoxicity caused by Nux Vomica.Conclusion:Through network toxicology analysis,it was found that lots of potential components in Nux Vomica may be involved in the activation of the excessive inflammatory response,oxidative stress,and the LPS response through multiple targets and multiple pathways,resulting in the generation of hepatotoxicity.

马钱子致大鼠体内毒性的研究——“装袋”算法和16S rRNA基因测序技术在毒理学研究中的应用

目的中药马钱子(Strychnos nux-vomica L.,SN)在临床上具有消肿止痛的功效,然而,由于含有生物碱类成分,马钱子具有一定毒性。人们对马钱子毒性所引起的大鼠内源性代谢变化及其对肠道微生物群代谢失调的潜在影响知之甚少,因此,马钱子的毒理学研究对其安全性评价具有重要意义。本研究将代谢组学和16S rRNA基因测序技术相结合来探索马钱子的致毒机制。方法通过急性、蓄积性和亚急性毒性试验,分别确定马钱子的中毒剂量、毒性强度和毒性靶器官。超高效液相色谱-质谱联用技术用于分析大鼠灌胃马钱子后的血清、肝脏和肾脏样本。利用基于装袋算法的决策树和K最近邻(K nearest neighbor,KNN)模型对组学数据进行分类。从大鼠粪便中提取样本后,使用高通量测序平台对细菌的16s rRNA V3-V4区域进行分析。结果装袋算法提高了样本分类的准确率。共鉴定出12个生物标志物,这些生物标志物的代谢失调可能是马钱子致体内毒性的原因。拟杆菌、粪厌氧棒菌、颤螺菌、双茎体菌等与肾肝功能的生理指标密切相关,这表明马钱子引起的肝肾损害可能与这些肠道细菌的代谢紊乱有关。结论本文揭示了马钱子的体内致毒机制,为马钱子临床上的安全合理使用提供了科学依据。

基于“络以通为用”探讨马钱子在特发性肺纤维化治疗中的应用

特发性肺纤维化属于“肺痹”“肺痿”范畴,其发病核心病机为顽痰瘀血阻于肺络,导致肺络痹阻不通。叶天士提出“络脉以通为用”,在特发性肺纤维化急性发作期应用通络药物可疏通肺络,减轻急性发作。马钱子走窜通络,善剔除沉积于肺络中痰瘀之邪。李光熙认为,马钱子虽有毒性,但其疏通络脉之力迅猛,临床多用于治疗痰瘀阻络证,在安全剂量内应用马钱子治疗特发性肺纤维化急性发作可明显改善患者临床症状,降低发作频率。从“络以通为用”论述特发性肺纤维化急性发作的病机,并从中西医角度讨论马钱子对于特发性肺纤维化急性发作的作用机理及临床应用马钱子的注意事项,以其为临床治疗特发性肺纤维化提供新的思路。

Cytotoxicities of alkaloids from processed and unprocessed seeds of Strychnos nux-vomica

目的：测定６种中药马钱子生品及不同炮制品的总生物碱以及１３个单体生物碱的细胞毒性．方法：采用细胞培养法观察马钱子生物碱对Ｖｅｒｏ细胞增殖抑制和［３Ｈ］胸腺嘧啶核苷酸参入法对宿主细胞内ＤＮＡ合成的影响．结果：砂炒炮制品的ＩＣ５０值分别是生品的１５５％和２１２％．１３个化合物的ＩＣ５０值分别是０．４５－０．８０ｍｍｏｌ·Ｌ－１和０．５０－１２ｍｍｏｌ·Ｌ－１．马钱子中异型生物碱及氮氧化合物在砂炒炮制品中含量最高，但这些生物碱显示出较小的细胞毒性．其中异马钱子碱氮氧化物毒性最低．砂炒炮制品与其它传统炮制品及生品比较具有更宽的安全范围．结论：马钱子的炮制对其毒性起关键性作用．

Zebrafish model for assessing induced organ toxicity by Strychnos nux-vomica

OBJECTIVE:To assess the acute organ toxicity of Strychnos nux-vomica with zebrafish model visually.METHODS:To assess acute toxicity,we initially determined the lethal concentration after Strychnos nux-vomica treatment for 24 h.Zebrafish was treated with five concentrations Strychnos nux-≦vo LC10 for 24 h,and the effects ofmica on morphology,function of heart,central nervous system,liver,kidney and organ-specific cell death were assessed.Next,we assessed the reversibility of toxic effect.RESULTS:Strychnos nux-vomica has an effect on the different organs of zebrafish,including heart,central nervous system,liver,and kidney,and cadiotoxicity induced by Strychnos nux-vomica was reversible to some extent.CONCLUSION:Zebrafish model is suitable for confirming the toxic target organs for Chinese traditional medicine.

Effect Evaluation of Strychnos nux-vomica L.with Integrative Methods for Bortezomib-Induced Peripheral Neuropathy in Multiple Myeloma Patients:A Self-Controlled Clinical Trial

Objective:To explore the clinical effect and adverse reactions of Strychnos nux-vomica in bortezomib-induced peripheral neuropathy(BIPN)of patients with multiple myeloma(MM).Methods:A total of 19 MM patients with BIPN were enrolled and Nux Vomica Capsule(NVC,0.4 g,thrice daily)were orally administrated for 30 days.Comparative analysis on parameters between pre-and post-therapy,including peripheral neuropathy(PN)grade,neurotoxicity score,Chinese medicine(CM)syndrome score,total neuropathy score(TNS),coagulation function,and serum nerve growth factor(NGF)levels were conducted.The adverse events were monitored.Results:In BIPN of MM patients who received NVC,PN grade was lowered,neurotoxicity score was obviously decreased(P 0.01),and both CM syndrome score and TNS were remarkably decreased(P<0.01).After the therapy,activated partial thromboplastin time was prolonged(P<0.01)and fibrinogen was declined(P<0.05),showing improvement in the hypercoagulable state of patients.No significant difference of NGF recovery degrees was detected between pre-and post-therapy(P>0.05).No evident adverse reactions were observed during the course of treatment.Conclusion:Strychnos nux-vomica L.has significantly effect with a good safety in treatment of BIPN in MM patients.

马钱子胶囊治疗缺血性脑卒中后认知功能障碍的疗效

目的观察马钱子胶囊治疗缺血性脑卒中后认知功能障碍的疗效。方法回顾性分析2020年1月至2022年12月90例急性期和恢复期的缺血性脑卒中患者的临床资料,观察组45例予马钱子胶囊联合常规治疗,对照组45例仅予常规治疗,比较急性期和恢复期两组患者治疗前后NIHSS、mRS、ADCS-ADL评分及综合疗效。结果急性期、恢复期两组治疗后NIHSS、mRS、ADCS-ADL评分优于治疗前(P<0.05);观察组NIHSS、mRS、ADCS-ADL评分治疗前后差值优于对照组(P<0.05)。结论观察组能改善缺血性脑卒中后认知功能障碍,但其机制仍待进一步探讨。

基于遗传算法优化绿豆煮马钱子的加压炮制工艺

目的优选绿豆煮马钱子的加压炮制工艺。方法采用最小二乘法建立炮制时间、炮制压力、绿豆用量和加水量4个因素对士的宁、马钱子碱含量的一元模型函数,通过对一元模型函数进行分析提出多元模型假设;基于正交实验,利用遗传算法对模型中的未定系数进行求解,构建基于士的宁、马钱子碱含量的双目标优化模型,求解模型函数得到最佳工艺,并对其进行验证。结果最佳炮制工艺为:马钱子与绿豆在2.393 MPa饱和蒸汽压力下煮5.5 h,捞出;冲洗除去绿豆,并刮去马钱子皮毛,切顶刀片0.6mm厚;每500g马钱子用绿豆180g、水15L。结论优化后的加压炮制工艺稳定、可行,可为绿豆煮马钱子炮制工艺优化提供参考。

基于PI3K/Akt/mTOR通路探究马钱子总生物碱对类风湿性关节炎大鼠的作用及机制

目的:分析马钱子总生物碱对类风湿性关节炎(RA)大鼠的作用及对磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶点(mTOR)通路的影响。方法:50只Wistar大鼠随机分为对照组、RA组、RA+马钱子总生物碱组(100 mg·kg^(-1))、RA+PI3K抑制剂组(20 mg·mL^(-1)LY294002)、RA+马钱子总生物碱+PI3K激活剂组[100 mg·kg^(-1)马钱子总生物碱+20μg·kg^(-1)胰岛素样生长因子1(IGF-1)],除对照组外,其余各组均构建RA模型并根据剂量给药,计算每只大鼠关节评分,测定大鼠血清抗瓜氨酸化蛋白抗体(ACPA)、丙二醛(MDA)水平;苏木精-伊红(HE)染色观察评估大鼠膝关节滑膜组织病理,比较各组Mankin评分、炎细胞浸润数、软骨厚度;测定滑膜组织白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)、核转录因子(NF)-κB受体活化因子配体(RANKL)水平;蛋白免疫印迹(Western blot)测定大鼠滑膜组织PI3K/Akt/mTOR通路相关蛋白表达。结果:1)对照组大鼠关节软骨组织结构正常,表面光滑完整,腔隙内组织有良好的完整软骨细胞;RA组关节表面广泛侵蚀,形态无规则,软骨细胞明显丢失,炎性细胞大量浸润,关节软骨厚度低,细胞间基质显著丢失,滑膜中单核炎性细胞浸润可见;RA+马钱子总生物碱组、RA+PI3K抑制剂组呈现保护性体征,关节表面侵蚀降低,炎性细胞浸润减轻,关节软骨厚度增加;与RA+马钱子总生物碱组相比,RA+马钱子总生物碱+PI3K激活剂组关节软骨损伤加重。2)与对照组相比,RA组大鼠关节炎症评分、血清ACPA及MDA水平、Mankin评分、炎细胞浸润数、滑膜组织IL-1β、IL-6、TNF-α、RANKL升高(P<0.05),软骨厚度降低(P<0.05);与RA组相比,RA+马钱子总生物碱组、RA+PI3K抑制剂组大鼠关节炎症评分、血清ACPA及MDA水平、Mankin评分、炎细胞浸润数、滑膜组织IL-1β、IL-6、TNF-α、RANKL降低(P<0.05),软骨厚度升高(P<0.05);与RA+马钱子总生物碱组相比,RA+马钱子总生物碱+PI3K激活剂组大鼠关节炎症评分、血清ACPA及MDA水平、Mankin评分、炎细胞浸润数、滑膜组织IL-1β、IL-6、TNF-α、RANKL升高(P<0.05),软骨厚度降低(P<0.05)。3)与对照组相比,RA组大鼠滑膜组织磷酸化(p)-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR升高(P<0.05);与RA组相比,RA+马钱子总生物碱组、RA+PI3K抑制剂组大鼠滑膜组织p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR降低(P<0.05);与RA+马钱子总生物碱组相比,RA+马钱子总生物碱+PI3K激活剂组大鼠滑膜组织p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR升高(P<0.05)。结论:马钱子总生物碱可能通过抑制PI3K/Akt/mTOR通路缓解RA大鼠炎症。

hERG钾通道介导马钱子碱和士的宁对癌细胞的增殖抑制作用

目的探讨马钱子碱和士的宁通过阻断hERG通道发挥抗肿瘤作用。方法采用脂质体转染及选择性培养基构建hERG转染CHO细胞和载体转染CHO细胞的稳定细胞株;CCK-8检测共培养24、48、72 h后马钱子碱及其结构类似物对癌细胞增殖的影响,以及共培养48 h后马钱子碱及其结构类似物对hERG-CHO细胞增殖的影响,以载体转染CHO细胞作为对照;流式细胞术检测马HT29、A549细胞周期;免疫印迹法检测HT29、PC9、A549细胞中hERG蛋白表达。结果马钱子碱和士的宁呈时间和浓度依赖性抑制HT29、PC9、A549细胞增殖,这种抗增殖作用与癌细胞中hERG蛋白表达正相关;而马钱子碱和士的宁的氮氧化物仅在高浓度对hERG通道有较弱的阻断活性,对肿瘤细胞也仅在高浓度表现出较弱的增殖抑制作用。马钱子碱对稳定表达hERG的CHO细胞的增殖抑制作用显著高于载体转染的CHO细胞。马钱子碱和士的宁将HT29细胞周期阻滞于G0/G1期,对A549细胞周期没有明显影响。马钱子碱不影响癌细胞的hERG蛋白表达。结论马钱子碱和士的宁对癌细胞的增殖抑制作用可能与hERG通道阻断相关,该研究将为hERG通道阻断剂在肿瘤治疗方面的应用提供重要的指导和依据。

马钱子中16个生物碱类化合物13CNMR谱的数据分析

从炮制前后的马钱子中，提取分离和鉴定了１６个生物碱，其中异马钱子碱、异马钱子碱氮氧化物、异番木鳖碱、氮氧化物、２－羟基，３－甲氧基番木鳖碱为新生物碱。经光谱（１３ＣＮＭＲ，１ＨＮＭＲ，ＩＲ，ＵＶ和ＭＳ）解析，确定其结构。通过实验发现，用１３ＣＮＭＲ谱测定马钱子中生物碱的结构，具有简便、快速等优点，且各化合物及其衍生物的共振峰呈现一定的规律。

马钱子不同炮制品中总生物碱的测定及急性毒性试验的比较

通过对中药马钱子不同炮制品中总生物碱的提取、含量测定、生物碱种类鉴别及小鼠急性毒性试验的比较，就炮制方法与毒性大小之间的关系进行了探讨。

马钱子生物碱在大鼠体内的组织分布

目的 研究马钱子生物碱在大鼠体内的组织分布。方法 大鼠静脉注射马钱子总碱后 ,定时用高效液相色谱法测定各组织中士的宁 (strychnine ,S)、马钱子碱 (brucine,B)、士的宁氮氧化物 (strychnineN oxide ,SNO)和马钱子碱氮氧化物(brucineN oxide ,BNO)的含量。结果 S、B、SNO和BNO在脑和脊髓中均有较多的分布。结论 S、B、SNO和BNO均可穿透血脑屏障 ,到达脑和脊髓 。

中药马钱子商品中士的宁和马钱子碱的含量测定

目的：对我国１５个省区产的中药马钱子商品及炮制品中的士的宁和马钱子碱进行含量测定。方法：应用反相高效液相色语法。结果：发现部分省市的生马钱子商品的士的宁含量偏高，个别炮制品的士的宁含量也超过了国家药典规定的标准。结论：因士的宁为剧毒成分，应引起足够的重视。

高效液相色谱法同时测定马钱子总生物碱脂质体中士的宁和马钱子碱的含量

目的：采用高效液相色谱法测定马钱子总生物碱脂质体中马钱子碱和士的宁的含量。方法：马钱子总生物碱脂质体与甲醇按1：9混合后离心取上清液用甲醇稀释进样。色谱条件：采用Lichrospher C18柱（4．6mm×250mm，5μm），乙腈-0．01mol·L^-1庚烷磺酸钠与0．02mol·L^-1磷酸二氢钾等量混合（用10％磷酸调pH值2．8）（24：76）为流动相，流速1．0mL·min^-1，检测波长254nm，柱温30℃，结果：马钱子碱、士的宁的线性范围均为1～20mg·L^-1，r=0．9999。低、中、高浓度士的宁平均加样回收率分别为（101．2±1．8）％，（99．2±1．3）％，（99．1±1．2）％（n=3）；低、中、高浓度马钱子碱的加样回收率分别为（103．9±2．8）％，（100．4±0．4）％，（102．7±1．2）％（n=3）。结论：本方法操作简便，是在药典方法基础上的改进，可用于控制马钱子总生物碱脂质体的质量。

马钱子总生物碱脂质体的含量与包封率测定

目的建立测定马钱子总生物碱脂质体含量和包封率的方法,用于处方与工艺评价。方法采用氨性氯仿法提取马钱子总生物碱并分析其成分,采用硫酸铵梯度法制备马钱子总生物碱脂质体。以士的宁为对照品,采用紫外分光光度法,在254nm测定用破膜剂溶解的马钱子总生物碱脂质体的吸光度确定其含量。采用葡聚糖凝胶柱分离脂质体和游离药物,分别测定含量后计算包封率。结果马钱子总生物碱提取物平均得率为3.49%,在总生物碱提取物中士的宁和马钱子碱分别占46.76%和26.51%。士的宁标准曲线的范围是2.4～20μg/mL,加样回收率在96.66～102.62%范围内。葡聚糖凝胶柱层析的回收率为100.85±0.79%(n=3)。结论所建立的方法简便快捷稳定,可以满足马钱子总生物碱脂质体质量控制的需要。

炮制对马钱子中生物碱的影响

应用薄层扫描法测定了按不同方法炮制的中药马钱子中士的宁、马钱子碱、异士的宁、异马钱子碱的含量，并就马钱子中生物碱的含量分布与其炮制方法之间的关系进行了研究。

不同炮制方法对马钱子中4种生物碱的影响

应用薄层扫描法测定了中药马钱子不同炮制品中４－羟基依卡精（ｖｏｍｉｃｉｎｅ）、依卡精（ｉｃａｊｉｎｅ）、士的宁氮氧化物（ｂｒｕｃｉｎｅＮ－ｏｘｉｄｅ）４种生物碱的含量，并就生物碱的含量分布与其炮制方法之间的关系进行了讨论。

爆压法对马钱子总生物碱的影响及急性毒性试验研究

目的 :观察爆压法对马钱子的总生物碱含量和急性毒性的影响。方法 :采用溶剂提取法提取总生物碱并测定其含量 ,用薄层层析对总生物碱中士的宁和马钱子碱进行定性分析 ,并通过小白鼠灌胃给药测定爆压品和生品的LD50 。结果 :马钱子经爆压法和砂烫法炮制后 ,总生物碱的含量下降甚微 ,与生品比较仍达 92 %～ 95 .6 % ,而爆压法与生品急性毒性比较下降了 38.3% (P <0 .0 1)。结论 :爆压法既能显著降低马钱子毒性 ,又能保证其效价不会明显下降。