Pyrrolizidine alkaloids-induced hepatic sinusoidal obstruction syndrome: Pathogenesis, clinical manifestations, diagnosis,treatment, and outcomes

Hepatic sinusoidal obstruction syndrome (HSOS) can be caused by the intake of pyrrolizidine alkaloids (PAs). To date, PAs-induced HSOS has not been extensively studied. In view of the difference in etiology of HSOS between the West and China, clinical profiles, imaging findings, treatment, and outcomes of HSOS associated with hematopoietic stem cell transplantation or oxaliplatin might be hardly extrapolated to PAs-induced HSOS. Reactive metabolites derived from PAs form pyrrole-protein adducts that result in toxic destruction of hepatic sinusoidal endothelial cells. PAs-induced HSOS typically manifests as painful hepatomegaly, ascites, and jaundice. Laboratory tests revealed abnormal liver function tests were observed in most of the patients with PAs-induced HSOS. In addition, contrast computed tomography and magnetic resonance imaging scan show that patients with PAs-induced HSOS have distinct imaging features, which reveal that radiological imaging provides an effective noninvasive method for the diagnosis of PAs-induced HSOS. Liver biopsy and histological examination showed that PAs-induced HSOS displayed distinct features in acute and chronic stages. Therapeutic strategies for PAs-induced HSOS include rigorous fluid management, anticoagulant therapy, glucocorticoids, transjugular intrahepatic portosystemic shunt, liver transplantation, etc. The aim of this review is to describe the pathogenesis, clinical profiles, diagnostic criteria, treatment, and outcomes of PAs-induced HSOS.

Genotoxic and tumorigenic pyrrolizidine alkaloids in Chinese herbal plants

Pyrrolizidine alkaloids are a class of hepatotoxic and tumorigenic compounds detected in Chinese herbal plants, contaminated foods, and dietary supplements. In this review, the sources, toxicity, genotoxicity, tumorigenicity, and the metabolic pathways, particular the activation pathways leading to hepatotoxicity and tumorigenicity, of pyrrolizidine alkaloids are briefly discussed, with a focus on the most recent important findings concerning the genotoxic mechanism by which riddelliine liver tumors. This mechanism involves the formation of 6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP)-derived DNA adducts and may be general to most carcinogenic pyrrolizidine alkaloids.

Recent advances on the synthesis of natural pyrrolizidine alkaloids: alexine, and its stereoisomers

Natural products have attracted the interest of the scientific community due to their importance and application.Alexine is a naturally polyhydroxylated pyrrolizidine alkaloid that is broadly found in plant sources and isolated from Alexa leiopetala.The biological properties such as glycosidase inhibitors,anti-virus,and anti-HIV activities,makes it interesting target for synthetical studies.This review reports different approaches and methodologies to the synthesis of alexine,and its stereoisomers as the target compounds in numerous studies.

Transjugular intrahepatic portosystemic shunt for pyrrolizidine alkaloid-related hepatic sinusoidal obstruction syndrome

BACKGROUND Treatments for hepatic sinusoidal obstruction syndrome(HSOS)are limited.AIM To evaluate transjugular intrahepatic portosystemic shunting(TIPS)as a treatment for pyrrolidine alkaloid-related HSOS(PA-HSOS).METHODS This retrospective analysis included patients with PA-HSOS admitted to the First Affiliated Hospital of the University of Science and Technology of China(June 2015 to January 2019).Baseline clinical characteristics and follow-up data were extracted from the medical records.All patients included in this study experienced failure of initial therapy.Patients were divided into the TIPS and conservative treatment groups according to the therapy they received.Liver function,maximal ascites depth,imaging characteristics,pathology findings,and survival were compared between groups.RESULTS The TIPS group included 37 patients(28 males),and the conservative treatment group included 17 patients(11 males).Baseline characteristics were similar between groups.There were two deaths in the TIPS group and seven deaths in the conservative treatment group during follow-up(3-48 mo).The 3-,6-,12-and 24-mo survival rates were 94.6%,94.6%,94.6%and 94.6%,respectively,in the TIPS group and 70.6%,57.8%,57.8%and 57.8%,respectively,in the conservative treatment group.Kaplan-Meier analysis revealed significantly longer survival for the TIPS group than for the conservative treatment group(P=0.001).Compared with the pre-treatment value,maximal ascites depth was significantly lower at 1 wk,2 wk,1 mo,and 3 mo for the TIPS group(all P<0.05)but not in the conservative treatment group.Contrast-enhanced computed tomography demonstrated the disappearance of patchy liver enhancement after TIPS.Pathology showed that liver congestion and hepatocyte swelling improved with time after TIPS placement.CONCLUSION TIPS may achieve better outcomes than conventional symptomatic treatment in patients with PA-HSOS.

滇紫草中肝毒性吡咯里西啶生物碱的含量分析

目的:建立液质联用法分析滇紫草中印美定、石松胺、蓝蓟定及其氮氧化物的含量。方法:采用沃特世HSS T3(长度100 mm、直径2.1 mm,粒径1.8μm)为色谱柱,以各含0.05%甲酸的乙腈水系统为流动相,梯度洗脱,流速为0.3 mL·min^(-1);进样量为1μL;电喷雾离子源,正离子多反应监测模式测定。结果:滇紫草中4种成分分别在0.972~97.2 ng·mL^(-1)、0.988~98.8 ng·mL^(-1)、1.024~102.4 ng·mL^(-1)、0.996~99.6 ng·mL^(-1)范围内线性关系良好(r>0.9990),回收率分别为93.5%、89.2%、88.1%和85.4%,RSD分别为3.5%、4.2%、3.2%和4.3%。结论:该方法专属性强,准确性好,可用于滇紫草中吡咯里西啶生物碱类成分的检测。

QuEChERS结合高效液相色谱-串联质谱法检测甘草中吡咯里西啶生物碱与风险分析

目的建立QuEChERS和高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)测定甘草中吡咯里西啶生物碱(pyrrolizidine alkaloids,PAs)的方法。方法甘草样品经0.05mol/L硫酸水QuEChERS自动样品制备系统振荡提取后,用混合型阳离子交换固相萃取小柱(PCX SPE)进行净化,以5 mmol/L甲酸铵水(含0.1%甲酸)-5 mmol/L甲酸铵甲醇为流动相,Agilent Proshell 120EC-C18柱进行分离,利用三重四极杆检测器在多反应监测模式下进行检测。结果利用建立的前处理方法对甘草中34种PAs做添加回收实验,在10、20和50μg/kg 3个浓度下的回收率为71.3%~112.0%,相对标准偏差为0.29%~8.18%,满足检测要求。利用建立的提取和检测方法对172个甘草样品中34种PAs进行监测,阳性样品检出率为22.1%,PAs总含量在9.5~118.0μg/kg之间。其中石松胺、石松胺N-氧化物、天芥菜碱和大尾摇碱N-氧化物的检出率最高。结论本研究开发的甘草中PAs的提取和检测方法快速、准确,可同时测定甘草中34种PAs阳性。所有监测的甘草样品中PAs总量低于欧盟限量标准。

九里明抗菌镇痛有效部位中PAs的研究

采用试管分项提取预试法、系统提取预试法和圆形纸层析预试法,进行了九里明植物化学成分预试;利用硅胶G薄板层析法,Ehrlich试剂喷雾显色,对九里明全草及其抗菌镇痛有效部位中双稠吡咯啶生物碱进行了检查。结果发现,九里明全草含有黄酮及其甙类、蒽醌及其甙类、酚类、强心甙、氨基酸、鞣质、皂甙、有机酸和生物碱,不含氰甙、多肽、蛋白质、内酯、油脂和挥发油;九里明全草含有双稠吡咯啶生物碱,但九里明抗菌镇痛有效部位中则未检出双稠吡咯啶生物碱。

茶叶中吡咯里西啶生物碱残留量检测方法及研究现状

综述了茶叶中吡咯里西啶生物碱(Pyrrolizidine alkaloids,PAs)残留量的检测方法,包括比色法、毛细管电泳法、免疫学方法、色谱分析法等,并总结了国内外茶叶中PAs残留研究现状,分析了茶叶中PAs的毒性作用以及国内外关于PAs的限量标准。我国目前尚缺乏茶叶中PAs的检测标准,迫切需要制定与欧盟规定相符的21种PAs的检测标准,并展开茶叶PAs污染水平监测与风险评估研究,为确保我国茶叶质量安全,积极应对欧盟贸易壁垒提供强有力的技术保障。

一体化QuEChERS净化-超高效液相色谱-串联质谱法测定茶叶中27种吡咯里西啶类生物碱

针对茶叶中吡咯里西啶类生物碱(Pyrrolizidine alkaloids,PAs)的残留风险,并为进一步提高传统QuEChERS的检测效率,研究建立使用一体化QuEChERS净化-超高效液相色谱-串联质谱测定茶叶中27种PAs的检测方法。该方法中的试样经1%甲酸乙腈提取,使用离心管一体盐包盐析和下压式QuEChERS净化,多反应监测模式(MRM)检测,基质匹配外标法定量。结果表明,27种PAs在各自浓度范围内线性关系良好,平均回收率为73.0%~111.8%,相对标准偏差为1.6%~13.8%(n=6)。该方法具有操作简单、检测通量大、灵敏度高、减少传统QuEChERS反复开盖等特点,能够满足茶叶中PAs的监测需求。对实际60批次茶叶样品进行检测,部分茶叶样品分别检出了不同含量的PAs,证明茶叶中有残留该类植物毒素的风险,建议持续跟踪监测。

SPE-UPLC-MS/MS法同时测定花草茶中的7种吡咯里西啶类生物碱及其氮氧化物

该研究建立了同时测定花草茶中7种吡咯里西啶类生物碱及其氮氧化物(野百合碱、野百合碱-N-氧化物、立可沙明碱、立可沙明碱-N-氧化物、天芥菜碱、天芥菜碱-N-氧化物、千里光宁碱)的固相萃取-超高效液相色谱-串联质谱(SPE-UPLC-MS/MS)分析方法。样品用0.1 mol/L盐酸溶液提取,经MCX固相萃取柱净化后,样液通过Poroshell 120 EC-C_(18)色谱柱(150 mm×3.0 mm,2.7μm)分离,电喷雾正离子多反应监测(MRM)模式检测。结果表明,7种吡咯里西啶类生物碱及其氮氧化物在1.0~50μg/L线性范围呈良好的线性关系,相关系数(R^(2))均>0.998;方法检出限(LOD)和定量限(LOQ)分别为0.3μg/kg和1.0μg/kg。平均加标回收率为81.8%~98.8%,精密度试验结果相对标准偏差(RSD)为1.98%~7.08%。对20个花草茶样品进行检测结果发现,其中有2个样品检出千里光宁碱,分别为18.2μg/kg和33.5μg/kg。该方法快速准确,准确度高,精密度好,适用于花草茶中的吡咯里西啶类生物碱及其氮氧化物的同时测定。

固相萃取-超高效液相色谱-串联质谱法测定菊花中15种吡咯里西啶生物碱毒素

目的建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)测定菊花中15种吡咯里西啶生物碱毒素含量的分析方法。方法样品中吡咯里西啶生物碱毒素采用0.1 mol/L硫酸水溶液提取,通过Oasis PRiME MCX固相萃取小柱净化,采用UPLC-MS/MS进行分析。采用Acquity UPLC BEH C18色谱柱分离,以0.1%甲酸水-甲醇溶液为流动相进行梯度洗脱,电喷雾离子源正离子电离,多反应离子监测模式监测,外标法定量。结果15种吡咯里西啶生物碱毒素在一定浓度范围内线性关系良好,相关系数(r2)均大于0.999;定量限为0.50~10.00μg/kg,检出限为0.15~3.00μg/kg;阴性样品的加标回收率为82.8%~92.4%,相对标准偏差为2.1%~4.9%(n=5)。结论该方法准确度和精密度高,耗时短,能同时测定多种菊花样品中的多种吡咯里西啶生物碱毒素含量。

固相萃取结合超高效液相色谱-串联质谱法快速测定乳粉及液体乳中26种吡咯里西啶生物碱的含量

以固相萃取结合超高效液相色谱-串联质谱技术,建立同时测定乳粉及液体乳中26种吡咯里西啶生物碱(pyrrolizidine alkaloids,PAs)含量的方法。样品采用5%三氯乙酸溶液进行提取、阳离子交换固相萃取柱净化后,经ACQUITY HSS T3色谱柱分离,以甲醇-0.1%甲酸溶液为流动相进行梯度洗脱,在电喷雾正离子源多反应监测模式下进行测定,采用外标法定量。在最优条件下进行方法验证,26种PAs化合物在1.0~40 ng/mL范围内线性关系良好,决定系数(R~2)均大于0.992,方法检出限(信噪比为3)为0.01~0.33μg/kg,定量限(信噪比为10)为0.03~1.0μg/kg。以空白乳粉和牛乳基质进行1、2、10倍定量限的加标回收实验(n=6),26种PAs化合物回收率在73.1%~111.6%之间,相对标准偏差为0.6%~9.8%。综上,该方法简单高效、灵敏度高、准确度好,适用于乳粉及液体乳中26种PAs的测定。

Validation of the Nanjing Criteria for Diagnosing Pyrrolizidine Alkaloids-induced Hepatic Sinusoidal Obstruction Syndrome

Background and Aims:Hepatic sinusoidal obstruction syndrome(HSOS)is caused by toxic injury to sinusoidal endothelial cells in the liver.The intake of pyrrolizidine alkaloids(PAs)in some Chinese herbal remedies/plants remains the major etiology for HSOS in China.Recently,new diagnostic criteria for PA-induced HSOS(i.e.PA-HSOS)have been developed;however,the efficacy has not been clinically validated.This study aimed to assess the performance of the Nanjing criteria for PA-HSOS.Methods:Data obtained from consecutive patients in multiple hospitals,which included 86 PA-HSOS patients and 327 patients with other liver diseases,were retrospectively analyzed.Then,the diagnostic performance of the Nanjing criteria and simplified Nanjing criteria were evaluated and validated.The study is registered in www.chictr.org.cn(ID:ChiCTR1900020784).Results:The Nanjing criteria have a sensitivity and specificity of 95.35%and 100%,respectively,while the simplified Nanjing criteria have a sensitivity and specificity of 96.51%and 96.33%,respectively,for the diagnosis of PA-HSOS.Notably,a proportion of patients with Budd-Chiari syndrome(11/49)was misdiagnosed as PA-HSOS on the basis of the simplified Nanjing criteria,and this was mainly due to the overlapping features in the enhanced computed tomography/magnetic resonance imaging examinations.Furthermore,most of these patients(10/11)had occlusion or thrombosis of the hepatic vein,and communicating vessels in the liver were found in 8/11 patients,which were absent in PA-HSOS patients.Conclusions:The Nanjing criteria and simplified Nanjing criteria exhibit excellent performance in diagnosing PA-HSOS.Thus,both could be valuable diagnostic tools in clinical practice.

Three new pyrrolizidine alkaloids derivatives from Liparis nervosa

Three new pyrrolizidine alkaloids, nervosineⅦ（1）, nervosine Ⅷ（2） and nervosine IX （3） were isolated from the whole plant extract of Liparis nervosa. Their structures were elucidated by extensive spectroscopic analyses （including 1 D, 2D NMR, and HR-ESI-MS） and chemical methods. Compounds 1-3 were evaluated for their cytotoxic activity against A549, MCF-7and H460 human cancer cell lines.

Pyrrolizidine alkaloids:An update on their metabolism and hepatotoxicity mechanism

Pyrrolizidine alkaloids(PAs)are among the most hepatotoxic natural compounds that are widely distributed throughout the world.Most PAs are metabolically activated to trigger toxicity.Exposure to herbal medicine containing PAs and food supplements contaminated by PAs is considered to be one of the two main causes of hepatic sinusoidal obstruction syndrome(HSOS),which is a rare hepatic vascular disease with a high mortality rate.PAs-induced HSOS cases have been reported worldwide.However,there is no clinically effective therapy for PAs-induced HSOS,which is partially because the toxic mechanism is not fully understood.This review focuses on updating the information on the metabolism and the molecular mechanisms of PAs hepatotoxicity,including oxidative stress,apoptosis,and dysfunction of bile acid metabolism,and their interactions.

Chemical Diversity Investigation of Hepatotoxic Pyrrolizidine Alkaloids in Qianliguang(Senecio scandens)and Related Species by UHPLC-QTOF-MS

Objective: Qianliguang(Senecio scandens) is a common Chinese medicinal herb. Qianliguang-containing herbal proprietary products are registered as over-the-counter remedies in China and exported to Western countries. The presence of hepatotoxic pyrrolizidine alkaloids(PAs)has raised concerns about the safety of using Qianliguang and its products. The present study aims at investigation of different types of PAs present in Qianliguang collected from representative locations in China.Methods: In this study, a simple but specific UHPLC-QTOF-MS method for the determination of toxic PAs was developed, based on the characteristic fragment ions specific to different types of PAs. It was successfully applied for the identification and distinguishing of PAs present in Qianliguang and related Senecio species growing in different locations of China.Results: Significant diversity of the PA types and quantities were revealed among the samples tested. The estimated total amounts of toxic PAs in three of the samples exceed the toxic limits of PA intake restricted by WHO, demonstrating the timely and highly demand for regulating both types and quantities of PAs present in Qianliguang.Conclusions: This study provides the methodology for simultaneous identification and quantification of PAs present in herbs without requiring corresponding standards, which could be further used for more systematic investigations of the PA distribution in Qianliguang and other PA-containing herbs.

苍术麸炒前后吡咯里西啶生物碱含量变化研究

目的:建立苍术中吡咯里西啶生物碱(PAs)的含量测定方法,考察麸炒前后5个PAs的变化规律。方法:采用UPLC-MS/MS法进行含量测定,采用ACQUITY UPLC HSS T3(150 mm×2.1 mm,1.8μm)色谱柱,流动相为乙腈-0.1%甲酸溶液(体积比为11∶89),恒流洗脱,流速0.30 mL·min^(-1),进样体积1μL;电喷雾离子源,正离子多反应监测模式,以标准曲线法计算含量。结果:3批苍术经过麸炒后,PAs总量从188.74~254.64μg·kg^(-1)下降到101.74~135.36μg·kg^(-1);野百合碱从25.35~52.18μg·kg^(-1)下降到23.25~37.41μg·kg^(-1);氮氧化野百合碱从25.12~40.22μg·kg^(-1)下降到6.98~18.23μg·kg^(-1);克氏千里光碱从11.64~36.25μg·kg^(-1)下降到3.64~12.85μg·kg^(-1);蓝蓟定从37.52~85.32μg·kg^(-1)下降到32.41~38.25μg·kg^(-1);氮氧化蓝蓟定从26.96~74.21μg·kg^(-1)下降到11.82~26.37μg·kg^(-1)。结论:该方法可用于苍术中PAs的测定。麸炒后PAs总量和单个含量均有降低,阐明了苍术麸炒解毒的科学内涵。

茶叶中21种吡咯里西啶生物碱检测方法研究

基于高效液相色谱-串联质谱建立了同时检测茶叶中21种吡咯里西啶生物碱(PAs)的分析方法。样品经0.05 mol/L硫酸提取,固相萃取法净化,Phenomenex Kinetex F5色谱柱(150 mm×3.0 mm×2.6µm)分离,以甲醇-0.1%甲酸水作为流动相进行梯度洗脱,正离子模式扫描,多反应监测(MRM)模式下进行检测,基质匹配标准曲线外标法定量。结果表明,21种PAs在各自质量浓度范围内呈良好线性关系(r^(2)≥0.9950),方法检出限(LOD)为0.3~1.0µg/kg;在低、中、高3个加标水平下,回收率为72.1%~108%,相对标准偏差(RSD,n=6)为2.0%~9.8%。将该方法用于市售茶叶样品的检测,有5件样品检出PAs,总含量在1.8~85µg/kg之间。所建立的方法高效、稳定,可同时实现21种吡咯里西啶生物碱的检测。

金丝桃苷改善吡咯里西啶生物碱诱导小鼠急性肝损伤的作用及机制研究

目的明确金丝桃苷对吡咯里西啶生物碱(PAs)诱导小鼠急性肝损伤的作用及机制。方法40只C57BL/6雄性小鼠随机分成5组,即空白对照组、模型组以及金丝桃苷低、中、高剂量组,每组8只。除空白对照组外,其余小鼠采用PAs建立急性肝损伤模型,造模6 h、30 h后金丝桃苷低、中、高剂量组小鼠分别灌胃给予20、40、80 mg/kg金丝桃苷各1次,造模48 h后,收集血清、肝脏、回肠、粪便。检测小鼠血清生化指标丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和总胆汁酸(TBA)水平;苏木精-伊红(HE)染色法观察小鼠肝脏组织病理学形态变化;测定小鼠血清、肝脏、回肠、粪便中胆汁酸含量;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法及Western blot法检测小鼠肝脏组织中法尼酯X受体(Fxr)、小异源二聚体伴侣(Shp)、胆盐输出泵(Bsep)、胆固醇-7α-羟化酶(Cyp7a1)、甾醇-12α-羟化酶(Cyp8b1)mRNA与蛋白表达水平。结果与空白对照组比较,模型组血清生化指标ALT、AST、TBA水平显著升高(P<0.05),肝细胞大面积坏死,同时伴有肝窦淤血;与模型组比较,金丝桃苷各剂量组血清生化指标ALT、AST、TBA水平显著降低(P<0.05),金丝桃苷中、高剂量组未见明显肝细胞坏死及肝窦淤血。与空白对照组比较,模型组胆汁酸代谢异常,小鼠肝脏Fxr、Shp、Bsep、Cyp7a1、Cyp8b1 mRNA及蛋白表达水平明显降低(P<0.05);与模型组比较,金丝桃苷中剂量组胆汁酸稳态失衡有所改善,小鼠肝脏Fxr、Shp、Bsep、Cyp7a1、Cyp8b1 mRNA及蛋白表达水平明显升高(P<0.05)。结论金丝桃苷可改善PAs诱导的急性肝损伤,其机制与调控胆汁酸相关基因、维护胆汁酸稳态平衡有关。

食物源吡咯里西啶类生物碱污染水平、检测技术及加工影响的研究现状

吡咯里西啶类生物碱(Pyrrolizidine alkaloids,PAs)是植物次生代谢产生的一类天然毒素,其中大部分不饱和型PAs及其氮氧化物(Pyrrolizidine alkaloid N-oxides,PANOs)具有多种毒性作用,并可通过膳食补充剂或食物链的形式进行传递从而威胁人类健康。PAs已被发现广泛存在于茶叶、蜂蜜等多种食品基质中,其污染成为令人担忧的食品安全问题。鉴于此,本文详尽梳理了不同国家不同食品基质中PAs的污染水平与可能的污染路径,综述了近年来PAs检测的前处理方法和分析检测技术的最新进展及发展趋势,分析了不同食品加工技术或烹饪制备对PAs污染物稳定性的影响,旨在为食品中PAs的检测研究、安全风险评估及控制提供警示和参考依据。