雷公藤肝毒性机制及配伍减毒研究进展

综合分析国内外雷公藤研究相关文献,对雷公藤临床不良反应及肝毒性表现进行表述和总结,分析肝毒性产生的机制。通过雷公藤配伍单味中药、有效化学成分及中药复方及炮制加工降低肝毒性进行归纳,并分析其降低毒性机制,从而明确雷公藤肝毒性研究现状及不足,并进一步对雷公藤药材及成分降低毒性提高疗效发展提出建议,以期为雷公藤药材研究发展及临床安全有效使用提供参考。     

雷公藤多苷肝毒性血清药物化学的初步研究

目的:通过对雷公藤多苷引起肝毒性入血成分的分析研究,探讨雷公藤潜在的肝毒性物质基础。方法:观察雷公藤多苷临床20倍等效剂量给药大鼠的肝组织病理形态学、血清AST和ALT来评价雷公藤多苷造成肝损伤的客观真实性;采用血清药物化学研究方法,在MSE质谱正负模式下,建立雷公藤多苷UPLC-Q/TOF-MS指纹图谱和其大鼠血清指纹图谱,比较确定雷公藤多苷的入血原型成分。结果:正常组与给药组大鼠肝组织病理切片指标、血清AST和ALT差异有统计学意义,血清指纹图谱的研究共鉴定Othosphenic Acid、Triptotriterpenic Acid、Demethylregelin、Wilforlide、Cangoronine 5个原型成分。结论:初步确定了雷公藤多苷引起肝毒性的入血成分,为其肝毒性物质基础的阐明奠定了基础。     

近20年来雷公藤肝毒性研究概述

雷公藤为治疗风湿病常用中药,疗效突出,然而其毒副作用,尤其肝毒性居单味肝损伤中药的首位,是困扰其临床运用的关键,也是阻碍其走向世界的绊脚石。基于此,该文对近20年来关于雷公藤肝毒性相关文献进行查阅,并从肝毒性表现、肝损伤的因素、毒性机制、减毒应对措施等方面对雷公藤的肝毒性研究进行总结,发现:①动物实验中雷公藤的肝毒性在生理状态下较病理状态下更为严重;②雷公藤的肝毒性是其所含有的多种有毒成分综合作用的结果,但生物碱类对肝脏毒性最大;用药剂量过大和累积服药量过大是雷公藤产生肝毒性的主要原因;③氧化应激学说仍是雷公藤肝毒性的重要机制,且Nrf2作为氧化应激的关键调控酶,已成为肝保护药物拮抗雷公藤肝毒性的作用靶点之一;线粒体自噬和肝超敏状态,是雷公藤诱导肝损伤的新机制;④控制用药量、药物配伍、炮制或改变剂型等措施有助于减轻雷公藤的肝毒性。该文旨在明确雷公藤肝毒性的研究现状和不足,以期提出新的研究策略,为合理评价雷公藤的安全性及临床合理用药提供思路。     

基于“异类相制”理论探讨雷公藤肝毒性配伍减毒的作用

雷公藤作为治疗类风湿性关节炎等自身免疫性疾病的首选药,其所含化学成分复杂,具有抗炎、抗肿瘤、免疫调节及抗生育等作用,是目前国内外研究的热点天然药物之一。但在使用中雷公藤所引起的肝损伤报道频繁发生,大大限制了其临床应用。依据"异类相制"的中药配伍理论,对雷公藤肝毒性减毒方面的有关文献进行归类分析,提出雷公藤肝毒性配伍减毒的组方原则,以期为提高雷公藤临床应用的安全性提供参考。     

雷公藤甲素肝毒性作用机制研究进展

雷公藤甲素是雷公藤的主要有效成分之一,近年关于其肝毒性的报道较多,笔者对近10年来关于雷公藤甲素所致肝毒性的作用机制的研究进行综述,以期为临床应用和科学研究提供参考依据。参考文献20篇。     

麒麟丸治疗雷公藤总苷引起的生殖毒性的理论探讨

雷公藤总苷是常用的免疫抑制剂,该药具有抑制免疫,抑制肾小球系膜细胞增殖,抑制炎症反应的作用,并且可以改善肾小球滤过膜的通透性,防止蛋白的丢失;然而雷公藤的副作用主要表现为性腺抑制,肝毒性和白细胞减少等,大大限制了临床的应用,文章通过探讨麒麟丸治疗雷公藤引起的生殖毒性的不良反应,为临床应用提供依据。     

雷公藤肝毒性作用机制研究进展

雷公藤系卫矛科藤本植物,具有抗炎、免疫调节等作用,临床应用广泛且疗效显著。雷公藤副作用涉及多器官多系统,尤以肝毒性报道最多,极大限制了其临床应用。从病理学基础、分子机制等方面对雷公藤致肝毒性的作用机制进行了阐述,以期为临床应用及研究提供理论依据。     

基于肝毒性的雷公藤中药复方配伍减毒的研究进展

随着中药毒理学研究的不断深入,雷公藤导致肝毒性的作用机制逐渐被揭示。中药配伍作为中医用药的特色之一,在减毒方面具有合理性和科学性。基于肝毒性,笔者对现阶段雷公藤毒性物质基础及机制的研究进行总结与展望,对配伍中药(制剂)以减轻雷公藤毒性的作用进行归纳,以期为雷公藤临床用药的合理性与安全性提供参考,从而减轻肝脏不良反应。     

雷公藤的肝毒性研究及ADME/Tox评价思路

雷公藤Triptergium wilfordii为卫矛科植物,具有多种药理作用,作为一种传统常用中药,其临床应用广泛,疗效显著,但使用中引起的肝损伤报道频繁发生.作为一种有毒中药,雷公藤本身所含化学成分多而复杂,且化合物多存在同物异名现象.现就雷公藤所含成分进行总结,并从相关临床病例报道出发,对雷公藤所致肝损害的特点及其可能的作用机制进行综述,提出基于ADME/Tox评价中药肝毒性的研究新思路,在原有的病理表征等传统研究方法的基础上,开展更为细致的中药毒性物质基础的研究.     

基于谱-效相关分析的雷公藤致肝毒性物质基础的初步研究

基于谱-效相关分析方法,筛选雷公藤致肝毒性的主要成分,为提升雷公藤类药材质量控制方法提供参考。该研究以中药雷公藤为研究对象,采用LC-Q-TOF-MS技术表征不同产地雷公藤样品的化学成分,并结合文献初步指认其主要成分;以正常人肝细胞(LO2细胞系)为模型,对乙酰氨基酚为阳性药,细胞毒价为评价指标,采用简单相关分析及多元线性相关分析方法,筛选雷公藤致肝毒性的主要成分。该研究共指认出雷公藤药材中的10种主要成分,基于谱-效相关分析发现雷公藤甲素可能是毒性成分,与传统文献报道一致,多元线性相关分析还发现雷公藤红素,demethylzeylasteral可能对肝毒性贡献较大;此外研究中还发现不同品种、不同产地雷公藤样品质量差异较大,产于西南地区的雷公藤药材肝毒性相对较低,产于湖南、安徽的2批雷公藤肝毒性相对较高。该研究可为今后雷公藤药材的合理利用及雷公藤致肝细胞毒性研究提供数据支持。     

维药芳香新塔花水提物的抗炎镇痛作用及其急性毒性

目的:研究维药芳香新塔花水提物的抗炎、镇痛作用及其急性毒性.方法:小鼠分别ig给予低、中、高剂量(0.5,1,2 g·kg-1)的芳香新塔花水提物,连续7d,1次/d.采用二甲苯致小鼠耳廓肿胀及甲醛致小鼠足跖肿胀的方法,观察其抗炎作用;以冰醋酸致小鼠扭体反应,观察其镇痛作用.3组小鼠分别ig给予7.5,15,30 g·kg-1的芳香新塔花水提物1次,进行急性毒性试验.结果:芳香新塔花水提物0.5,1,2 g·kg-1对二甲苯致小鼠耳廓肿胀、甲醛致小鼠足肿胀及醋酸致小鼠扭体均有显著抑制作用(P＜0.05);30 g·kg-1及以下剂量单次ig给药后小鼠全部存活,观察期内未见明显毒性反应.结论:芳香新塔花具有明显的抗炎镇痛作用.     

小叶金花草抗炎镇痛作用

目的:对小叶金花草水提取物和醇提取物的抗炎、镇痛作用及急性毒性进行研究。方法:采用二甲苯致小鼠耳廓肿胀法、醋酸扭体法观察小叶金花草的抗炎、镇痛作用;以小叶金花草水提物1.62 g·mL-1,醇提物2.34 g·mL-1,最大给药容积40 mL·kg-1分别对小鼠ig给药,观察其急性毒性。结果:小叶金花草水提取物和醇提取物10,5,2.5 g·kg-1对二甲苯所致小鼠耳肿胀有明显的抑制作用(P<0.05),抑制率分别为35.9%,32.5%,29.5%和31.0%,30.0%,24.6%;小叶金花草水提取物和醇提取物对0.7%醋酸所致小鼠扭体反应有明显的抑制作用(P<0.05);小叶金花草水提物对小鼠ig给药的最大耐受量为56.5 g·kg-1;醇提物对小鼠ig给药的最大耐受量为85.7 g·kg-1。结论:小叶金花草具有一定的抗炎、镇痛作用;小叶金花草水提物及醇提取物在所试最大耐受剂量未观察到明显毒性反应。     

雷公藤制剂指纹图谱及谱效关系研究

采用高效液相色谱法(HPLC)建立7个厂家雷公藤多苷片醋酸乙酯提取物的指纹图谱,得到11个共有峰,各厂家指纹峰种类和含量差异较大.选择Con A刺激的小鼠脾淋巴细胞模型进行药效学研究,分别给予30,15,7.5,3.75,1.88,0.94mg·L-1的雷公藤多苷片提取物进行处理,48 h后测定各药物对小鼠脾淋巴细胞增殖的作用,发现各厂家的雷公藤多苷片均对小鼠脾淋巴细胞增殖有显著抑制作用(P＜0.05),并呈剂量依赖性,且各厂家雷公藤制剂对小鼠脾淋巴细胞增殖的作用的IC50值差异较大.选择灰色关联度分析法分析指纹图谱与增殖抑制药效数据的关系,发现各指纹峰与药效的关联度均大于0.6,表明雷公藤增殖抑制药效是多成分共同起效的结果,其中5号峰与10号峰对增殖抑制作用贡献最大,各共有峰与药效之间关联序为5＞10＞8＞7＞6＞11 ＞4＞9＞2＞3＞1.该文初步探讨了雷公藤的物质基础,为明确雷公藤的作用机制,建立更加全面可靠的质量控制方法提供理论依据与数据支持.     

Evaluation of toxicity of Calophyllum brasiliense stem bark extract by in vivo and in vitro assays

Ethnopharmacological relevance

Calophyllum brasiliense Camb., Clusiaceae, is commonly known as “guanandi” and its stem bark is used in Brazilian traditional medicine to treat rheumatism, vein problems, hemorrhoids and gastric ulcers. The aim of this study was to evaluate the toxicity of hexane extract of Calophyllum brasiliense stem bark (HECb) using in vitro and in vivo experimental models.

Materials and methods

In vitro toxicity was evaluated by Alamar Blue cytotoxicity assay and micronucleus test, using Chinese hamster ovary (CHO-k1) epithelial cells. in vivo toxicity was evaluated by oral acute and subchronic toxicity assays. In the oral acute toxicity screening, a single dose of HECb was administered to mice at doses ranging from 250 to 1000 mg/kg. In the subchronic study, HECb was administered orally for 30 days to Wistar rats at doses of 100 mg/kg and 500 mg/kg. Phytochemical analyses were performed by HPLC/UV–vis, secondary metabolites were quantified by spectrophotometric methods.

Results

HECb presented IC₅₀=119.94±4.31 µg/mL after a 24 h cytotoxicity test using CHO-k1 cells, showing low cytotoxicity. However, when the cells were exposed to HECb for 72 h, the IC₅₀ value was 8.39±2.00 µg/mL, showing in this case, a pronounced cytotoxic effect. In the oral acute toxicity studies, doses up to 500 mg/kg of HECb did not cause any changes in both male and female mice. At 1000 mg/kg, male mice showed signs typical of depression and stimulation that were reversed at 72 h. Besides, female mice were more sensitive to the toxic effect of HECb at 1000 mg/kg, which initially presented typical agitation signals, followed by depression signals, leading to death of all the animals at 24 h. In subchronic assay with rats, HECb administered orally at doses of 100 and 500 mg/kg did not cause significant changes in all clinical parameters evaluated. Histopathological analyses showed no deleterious effect in the vital organs of rats. Preliminary phytochemical analysis revealed the presence of phenolic compounds, steroids, and volatile coumarins. Analysis by HPLC showed two major peaks characteristic of chromanones.

Conclusions

In vitro toxicological tests showed that HECb exhibited cytotoxicity especially after 72 h of exposition, and mutagenicity on the highest tested dose. The in vivo studies demonstrated that HECb produced some toxicity signs at the highest dose tested, particularly, in the acute toxicity test but showed no significant signs of toxicity in the subchronic assay. Based on these and previous pharmacological studies, it is possible to say that HECb did not exhibit significant toxicity at its effective dose. This suggests that HECb is relatively safe in humans at its effective dose.     

Evaluation of safety of the herbal formula Ojeok-san: Acute and sub-chronic toxicity studies in rats

Aim of the study

Ojeok-san (OJS; wuji powder in China and goshaku-san in Japan), a widely used herbal formula in traditional Korean medicine and Japanese herbal medicine (Kampo medicine), has been used to treat common cold and illnesses including fatigue and gastrointestinal disorders, but there is very little information on its safety. To provide information on the safety of OJS, we evaluated its acute and sub-chronic toxicity in rats.

Materials and methods

The single and sub-chronic toxicity of OJS was examined using male and female Sprague-Dawley rats. The rats were treated with the OJS extract orally at the highest dose level of 2000 mg/kg/day body weight. After single administration, signs of toxicity were observed every hour for the first 6 h and every day for two weeks. In the sub-chronic toxicity study, OJS was administered for 13 weeks. Mortality, clinical signs, body weight changes, food and water consumption, ophthalmologic findings, urinalysis, hematological and biochemical parameters, gross findings, organ weights and histological markers were monitored during the study period.

Results

We found no mortality and no abnormality in clinical signs, body weight, and necropsy findings for any of the animals in the acute and sub-chronic toxicity study following oral administration of OJS.

Conclusion

OJS may not have any single dose toxicity. The lethal dose with a 50% mortality rate (LD₅₀) was over 2000 mg/kg. The no-observed adverse effects level (NOAEL) was considered to be 2000 and 1000 mg/kg/day for male and female rats, respectively.     

Safety assessment of aqueous extract from leaf Smallanthus sonchifolius and its main active lactone, enhydrin

Ethnopharmacological Relevance

Leaves of Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson (yacon) have been used since pre-Columbian times in the Andean region to prepare medicinal herbal tea with beneficial health properties. However, there are still disagreements about the safe use. This work was carried out to evaluate the toxicity profile of both, 10% decoction of yacon leaves and their major active lactone, enhydrin.

Materials and methods

In vitro cytotoxicity assays were performed with Hep-G2, COS1, CHO-K1 and Vero cell lines using a test of metabolic competence based upon assessment of mitochondrial performance. In vivo toxicity study was performed in adult Wistar rats. In the acute oral toxicity each group of rats was orally given a single dose of 10% decoction or enhydrin. General condition, behavior and mortality were recorded for up to 14 days post treatment. In subchronic toxicity studies, both products were given orally for 90 days to rats. Body weight and food intakes were observed weekly. Hematological, clinical chemistry parameters and organ weight were determined in all animals at the end of the experimental period.

Results

Cell viability decreased in a concentration dependent fashion when cells were incubated with 2–200 μg of 10% decoction and 0.015–7.5 μg of enhydrin. In acute study in rats, there were no deaths or signs of toxicity observed after oral administration of single doses of 10% decoction or enhydrin at any dose level up to the highest dose tested (14.0 g/kg and 0.32 g/kg, respectively). In subchronic studies in rats, both products administered orally for 90 days at daily doses of 0.07, 0.14 and 0.28 g 10% decoction/kg and 0.4, 0.8 and 8.0 mg enhydrin/kg, did not caused haematological, biochemical and histological alterations.

Conclusions

The results presented in this paper lead us to the conclusion that the use of 10% decoction and enhydrin is safe in rat at doses in which it is demonstrated the hypoglycaemic effect.     

Acute and subchronic oral toxicity of Coriolus versicolor standardized water extract in Sprague-Dawley rats

Ethnopharmacological relevance

Coriolus versicolor, which is known as Yun Zhi, is one of the commonly used Chinese medicinal herbs. Recent studies have demonstrated its antitumor activities on cancer cells which led to its widespread use in cancer patient. However, little toxicological information is available regarding its safety. The present study evaluated the potential toxicity of Coriolus versicolor standardized water extract after acute and subchronic administration in rats.

Materials and methods

In acute toxicity study, Coriolus versicolor water extract was administered by oral gavage to Sprague-Dawley (SD) rats (6 males, 6 females) at single doses of varying concentrations 1250, 2500 and 5000 mg/kg. In subchronic toxicity study, the extract was administered orally at doses of 1250, 2500 and 5000 mg/kg/day for 28 days to male and female SD rats respectively. General behavior, adverse effects and mortality were determined throughout the experimental period. Haematological and biochemical parameters, relative organ weights and histopathological were evaluated at the end of the experiment.

Results

There were no mortality and signs of toxicity in acute and subchronic toxicity studies. In the single dose acute toxicity and repeated dose 28-day subchronic toxicity studies, there were no significant difference in body weight, relative organ weight, haematological parameters, clinical chemistry, gross pathology and histopathology between treatment and control groups.

Conclusions

Coriolus versicolor water extract did not cause remarkable adverse effect in SD rats. The oral lethal dose of Coriolus versicolor water extract is more than 5000 mg/kg and no-observed-adverse-effect level (NOAEL) of the extract for both male and female rats is 5000 mg/kg per day for 28 days.     

Repeated-dose toxicological studies of Tithonia diversifolia (Hemsl.) A. gray and identification of the toxic compounds

Ethnopharmacological relevance

Tithonia diversifolia (Hemsl.) A. Gray has been commonly used in folk medicine to treat abscesses, microbiological infections, snake bites, malaria and diabetes. Both anti-inflammatory and anti-malarial properties have been identified using appropriate assays, but the effective doses have demonstrated toxic effects for the experimental animals. Most of the pharmacological activities have been attributed to sesquiterpene lactones (STLs) and some chlorogenic acid derivatives (CAs) in the leaves of this species. This work aimed to evaluate the repeated-dose toxicity of an aqueous extract (AE) from Tithonia diversifolia leaves and to compare the results with an extract rich in STLs (LRE) and a polar extract (PE) without STLs but rich in CAs. The purpose of this work was to provide insights into the identity of the compounds responsible for the toxic effects of Tithonia diversifolia.

Materials and methods

The major classes of compounds were confirmed in each extract by IR spectra and HPLC-UV-DAD profiling using previously isolated or standard compounds. The toxicity of each extract was evaluated in a repeated-dose toxicity study in Wistar rats for 90 days.

Results

The AE is composed of both STLs and CAs, the LRE is rich in STLs, and the PE is rich in CAs. The AE caused alterations in haematological parameters but few alterations in biochemical parameters and was relatively safe at doses lower than 100 mg/kg. However, the PE and LRE demonstrated several adverse effects by damaging the liver and kidneys, respectively.

Conclusion

STLs and CAs can be toxic in prolonged use at higher doses in extracts prepared from Tithonia diversifolia by affecting the kidneys and liver.     

雷公藤配伍减毒研究进展

雷公藤具有清热解毒、祛风除湿、消肿止痛、杀虫止痒的功用,现代药理研究亦表明其具有抗炎、镇痛、抗肿瘤及免疫调节等作用,临床主要用于治疗免疫性疾病、肾脏性疾病、皮肤性疾病等,对治疗类风湿性关节炎疗效尤为显著。但因其具有严重的毒副反应,尤其是肝肾毒性,限制了雷公藤的临床安全应用,成为了迫切需要解决的难题。虽然目前存在的减毒方式众多,包括传统的净制减毒、炮制减毒、配伍减毒和现代的制剂减毒、结构修饰减毒、生物技术减毒以及其相互间的联合应用等,但并没有本质上解决雷公藤的安全问题。在古代医药文献中药物的毒性常指这种药物的偏性,在中医药理论看来,作为药物都有毒性,关键是正确使用。中药的配伍使用是中医药文化的重要组成部分,从春秋战国时期的配伍雏形,到《黄帝内经》中已有君臣佐使配伍用药的方剂,再到《神农本草经》中配伍理论的形成,到最后《伤寒论》经典名方的传世,记录了中药配伍应用的发展,也记录了中药配伍使用的重要意义,随着中药事业逐渐走向现代化和国际化,中药配伍又有了进一步的发展,开始迈向质量可控、机制清楚的组分配伍,故本文以雷公藤配伍减毒为中心,以配伍形式的发展为主线,对雷公藤配伍减毒进行综述,为雷公藤减毒增效研究提供思路,促进其合理开发和临床安全应用,也为其他有毒中药的合理应用提供参考。