Advances in toxicological studies of Radix Phytolaccae

Radix Phytolaccae is the dried root of Phytolacca acinosa Roxb or P.ameri-cana L,which is commonly used as a traditional Chinese medicine to treat diseases like cirrhotic ascites,hepatitis B,nephrotic syndrome,psoriasis,etc.However,there is no exact basis for its clinical application safety.In this paper,the toxic effects and mechanism of Saponin A(EsA),the main component of Radix Phytolaccae,were summarized by searching the results and reports of toxicology related to the plant from 1991 to 2023 on CNKI and pubmed,aiming to provide reference for the toxicological research and future research direction of Radix Phytolaccae,so that Radix Phytolaccae can be safely and effectively used in clinical practice.

商陆(Phytolacca acinosa Roxb.)的锰耐性和超积累

超积累植物的确证对成功实施重金属污染环境植物修复是必不可少的。通过野外调查和营养液培养试验 ,研究超积累植物商陆 (PhytolaccaacinosaRoxb )的锰富集特性 ,结果表明 ,商陆对生长介质中的Mn具有很强的耐性和累积能力。商陆在锰含量高达 114× 10 3 mgkg-1的尾矿废弃地上依然生长良好 ,叶锰含量最高达 19 3× 10 3 mgkg-1。温室培养条件下 ,当生长介质中Mn浓度为 8 0 0 0mmolL-1时 ,虽然其生物量与对照相比有所降低 ,但植株仍能生长。随着生长介质中Mn浓度的升高 ,商陆叶和茎的Mn含量逐渐增加 ,生物富集系数则逐渐降低 ,但是地上部分锰积累量则先增加后减少。当Mn浓度为 5 0 0 0mmolL-1时 ,商陆地上部分锰积累量达到最大值 2 5 8 2mgplant-1;当Mn浓度为 12 0 0 0mmolL-1时 ,商陆仍能完成整个生命周期 ,叶锰含量达到最大值 36 4× 10 3 mgkg-1,生物富集系数为 5 5。不同锰供应水平下 ,商陆吸收的锰有87%～ 95 %被转移到地上部分。这进一步验证商陆的锰超积累特性 ,为利用超积累植物对大面积污染土壤实施植物修复提供了有力证据 。

锰超富集植物垂序商陆(Phytolacca americana L．)的认定

针对目前国内锰超富集植物商陆属(Phytolacca)植物名称混乱的状态,在重新审查锰超富集植物——商陆(Phytolacca acinosa Roxb.)的野外标本和温室培养植物的基础上,比对中国科学院昆明植物所标本馆的腊叶标本,并查阅相关文献资料,以期正确认定锰超富集累植物的学名。结果表明,湘潭锰矿尾矿废弃地原生的锰超富集植物实为垂序商陆(Phytolacca americana L.),国内外相关研究论文中出现的商陆和美洲商陆实为垂序商陆的同物异名。这一植物名称的认定,将对避免锰超富集植物研究重复进行、保证相关研究正常有序开展具有重要意义。

锰胁迫对商陆(Phytolacca acinosa)保护酶的影响

对室内栽培的商陆(Phytolacca acinosa)幼苗分别经4、12、36 mmol/L的锰(Mn2+)溶液处理30d后,用聚丙烯酰胺凝胶电泳(PAGE)分离了根和叶片过氧化物酶(POD)同工酶和超氧化物歧化酶(SOD)同工酶,并测定了其酶活性。结果表明:(1)经不同浓度的Mn2+溶液处理,根POD同工酶(RP1)消失,而12、36 mmol/L Mn2+溶液处理根新的POD同工酶(RP5,RP8)出现,根POD酶活性出现先降低后升高的变化趋势;叶片POD同工酶(LP)有3种,与对照的相同,但其酶活性随Mn2+浓度增加而随之升高。(2)12、36 mmol/L Mn2+溶液处理,根新的SOD同工酶(RS5-RS7)出现,SOD酶活性上升;36 mmol/L Mn2+溶液处理,叶片SOD同工酶(LS2-LS5)消失,叶片SOD酶活性下降。推断,商陆根和叶片同工酶的表达调控的机制不同。认为,商陆在Mn2+胁迫中,POD发挥更重要的作用。

美洲商陆(Phytolacca americana L.)——一种新的Mn积累植物

通过野外调查和水培试验,研究了锰在美州商陆体内积累情况及对其生长发育的影响,发现并鉴定出美洲商陆(Phytolacca am ericana L.)是一种新的M n积累植物。调查结果发现,美洲商陆对土壤中高含量的锰有很强的忍耐、吸收和积累能力,叶片中M n含量为5160￣8000m g·kg-1,平均为6490m g·kg-1。营养液培养试验证明,美洲商陆对生长介质中的M n有很强的忍耐能力,当生长介质中M n浓度高达10m m ol·L-1,叶片中M n含量达到11.76g·kg-1,植株仍生长正常。地上部M n含量及其积累量均随生长介质中M n浓度的增加而增加,当生长介质中M n浓度为50m m ol·L-1时,叶片中M n含量达到47.06g·kg-1。结果表明,美洲商陆是我国发现的一种新的M n积累植物,为今后探明植物积累M n的机理和M n污染土壤的植物修复提供了一种新的种质资源。

An investigation of cellular distribution of manganese in hyperaccumlator plant Phytolacca acinosa Roxb. using SRXRF analysis

Phytolacca acinosa Roxb. （P acinosa） is a recently discovered manganese hyperaccumulator plant from southern China. It is a good candidate for phytoremediation of manganese（Mn） polluted soil for its high biomass and fast growth. Knowledge of the tissue localization and identification of heavy metals can provide essential information on metal toxicity and bioaccumulation mechanisms. Synchrotron radiation X-ray fluorescence spectroscopy （SRXRF） microprobe was used in this study to investigate the cellular distributions of Mn and other elements in root, stem, leaf, petiole and midrib of P. acinosa. The highest Mn content was found in the vascular tissues of root, stem, petiole and midrib. Cortex in root played a key role in Mn absorption and Mn was limited in the vascular bundle during the process of transportation in stem. Moreover, Mn content in leaf epidermis was higher than that in mesophyll, which suggested that the sequestration of Mn in leaf epidermis might be one of the detoxification mechanisms of P. acinosa. The significance of other elemental （such as P, S, K, Ca, Fe, Zn and Cu） distribution patterns and the correlation with Mn were also discussed.

Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation

OBJECTIVE： Preventive measures against skin melanoma like chemotherapy are useful but suffer from chronic side effects and drug resistance. Ethanolic extract of Phytolacca decandra （PD）, used in homeopathy for the treatment of various ailments like chronic rheumatism, regular conjunctivitis, psoriasis, and in some skin diseases was tested for its possible anticancer potential. METHODS： Cytotoxicity of the drug was tested by conducting 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide assay on both normal （peripheral blood mononuclear cells） and A375 cells. Fluorescence microscopic study of 4＇,6-diamidino-2-phenylindole dihydrochloride-stained cells was conducted for DNA fragmentation assay, and changes in cellular morphology, if any, were also recorded. Lactate dehydrogenase activity assay was done to evaluate the percentages of apoptosis and necrosis. Reactive oxygen species （ROS） accumulation, if any, and expression study of apoptotic genes also were evaluated to pin-point the actual events of apoptosis. RESULTS： Results showed that PD administration caused a remarkable reduction in proliferation of A375 cells, without showing much cytotoxicity on peripheral blood mononuclear cells. Generation of ROS and DNA damage, which made the cancer cells prone to apoptosis, were found to be enhanced in PD-treated cells. These results were duly supported by the analytical data on expression of different cellular and nuclear proteins, as for example, by down- regulation of Akt and Bcl-2, up-regulation of p53, Bax and caspase 3, and an increase in number of cell deaths by apoptosis in A375 cells. CONCLUSION： Overall results demonstrate anticancer potentials of PD on A375 cells through activation of caspase-mediated signaling and ROS generation.

Evaluation of berries of Phytolacca dodecandra for growth inhibition of Histoplasma capsulatum var.farciminosum and treatment of cases of epizootic lymphangitis in Ethiopia

Objective:To evaluate the berries of Phytolacca dodecundra(P.dodecandra) for its effect on Histoplasma cupsulatum var.farciminosum(HCF) and for the treatment of cases of epizootic lymphangitis(ELi.Methods:Samples were collected from un-ruptured nodules of cases of EL at Debre Zeit and Akaki(central Ethiopia).Mycologieal culture and isolation of HCF were performed at the Akliln Lemma Institute of Pathobiology.Phytochemical screening was done for n-butanol extract of P.dodecandra to delect alkaloids,saponins,phenolic compounds and flavonoids.The minimum inhibitory concentrations(MICs) and minimum fungicidal concentrations(MFCs) ol aqueous and n-butanol extracts of P.dodecandra against FICF were determined by agar dilution assay.For the in vivo trial.5%simple ointment was prepared from n-butanol extract and applied topically to 24(twelve early and twelve moderate) cases of F.L.Results:Phytochemical screening showed that n-butanol extract ol P.dodecandra was positive lor alkaloids.saponins and phenolic compounds but negative for flavonoids.The MFCs of n-butanol and aqueous extracts of P.dodecandra were(0.039%-0.078%) and(0.625%-1.250%),respectively.The MFCs of n-butanol and aqueous extracts of P.dodecandra were(0.078%t-0.156%)and(1.250%-2.500%),respectively.The MIC and MFC of ketoconazole(positive control) was(1.200×10^(-5)%-2.500×10^(-5)%) and(5.000× 10^(-5)%-1.000×10^(-4)%),respectively while growth was observed on free medium(negative control).From the total of 24 treated cases of EL,14(58.3%) responded lo treatment;however,10(41.7%) did not respond to treatment.There was no significant difference in the degree of response to treatment between early and moderate cases(χ~2=0.086:P=0.408.Conclusions:It can be concluded that n-butanol extract of P.dodecandra demonstrates antifungal effects while the aqueous extract shows no antifungal activity.

Study on Induction of Phytolacca acinosa Roxb. Polyploids with Colchicine

[ Objective ] This study aimed to obtain Phytolacca acinosa Roxb. polyploids and provide high-efficiency raw materials for the preparation of biological pesticides. [Method] Terminal bud growing points of Phytolacca acinosa Roxb. seedlings were treated with 0.1%, 0.2%, 0.3% colchicine and 1% agar solution for 24, 48 and 72 h, respectively, while Phytolacca acinosa Roxb. without treatment was adopted as the control. The mutagenic effect of various concentrations of colchicine was calculated. Phytolacca acinosa Roxb. mutants were identified using morphological, anatomical and cytological methods. [ Result] Induction with 0. 2% colchicine for 48 h led to the highest mntagenic effect. Compared with the control, leaf width and leaf thickness of mutants were enhanced by 40. 15% and 33. 25%, respectively; vertical and horizontal diameter of stomata of mutants were enhanced by 30.25% and 60.82%, respectively ; the stomata density of mutants was improved by 150%. Identification of meiotic chromosome of pollen mother cells using compression method showed that the chromosome number of microspore cells of Phytolacca acinosa Roxb. mutants was n = 2 x = 36, while that of the control was n = x = 18, indicating that the mutants were tetraploids. [ Conclusion ] Phytolacca acinosa Roxb. polyploids were induced using the traditional liquid droplet method, which was easy and direct with simple operation and good effect to acquire a large number of polyploid materials within a short time.

不同生境美洲商陆(Phytolacca americana L.)根系差异表达蛋白筛选

某铀尾矿区生长着大量美洲商陆（Phytolacca americana L.）,目前还未见美洲商陆耐受及富集铀相关蛋白质组学研究报道。为了找到铀耐受及富集表达相关蛋白,建立了铀尾矿区和对照区2种不同生境美洲商陆根系蛋白质组研究技术。通过全蛋白质制备条件优化、2DE电泳条件摸索以及染色方法选择,得到了重现性好、分离效果和清晰度都很高的美洲商陆根系双向电泳差异凝胶图谱。经软件分析,筛选出了铀耐受及富集相关蛋白24个,上调表达11个、下调表达13个。

Floral Organogenesis and Ring Meristem in <i>Phytolacca</i>

To further study the floral organogenesis and discussing the floral origin of Phytolacca, the procedures of floral organogenesis were observed in Phytolacca esculenta and Phytolacca zhejiangensis. The results showed that the floral organogenesis was consistent in Phytolacca. Their sepals were 2/5 helix, and with counter-clockwise and clockwise, usually the first sepal located at non-median of abaxial side. The first sepal of Phytolacca esculenta was initiated at non-median of adaxial side. There was no evident relationship between sepal and stamen initiating position, and the stamens initiated on ring meristem, they initiated approximately at the same time, and when the androecium member was numerous, they initiated centrifugally, the outer stamen initiated irregularly. Carpel initiated alternately with inner stamens. And the carpels connected by septum, if the septum grew more, the carpel was syncarpous at morphology, otherwise the carpel was apocarpous at morphology. So the syncarpous and the apocarpous have no successively relationship on evolution. Ovule initiated inside the carpel and opposite to carpel. Androecium, carpel and ovule initiated at ring meristem.

Artificial Triterpenoid Fatty Acid Ester Isolated From the Leaves of Phytolacca icosandra L

The methanol extract form the leaves of Phytolacca icosandra L.,afforded the unprecedented artificial triterpenoid fatty acid ester 1 derived from the new natural triterpenoid phytolaccagenic acid 3-O-myristate(1a),along with the three known triterpenoids serjanic,acinosolic and phytolaccagenic acid(2-4).Their structures were stablished by HR-EI-MS,1D and 2D NMR techniques.The possible mechanistic formation of 1 is proposed,and the in vitro toxicity of all compounds was assessed using the brine shrimp lethality assay(BSLA).

Analgesic and Anti-Inflammatory Activity of Total Crude Leaf Extract of <i>Phytolacca dodecandra</i>in Wistar Albino Rats

Background: Pain is a common problem encountered in daily life among humans and animal and it is commonly managed conventionally using modern medicines and traditionally by use of medicinal plants. Phytolacca dodecandra is a common herb in Uganda, used traditionally to alleviate pain and inflammatory conditions. However, its activity in alleviating pain and inflammatory diseases has not been fully scientifically evaluated. Aim of Study: To evaluate the analgesic and anti-inflammatory activity of total crude leaf extracts of P. dodecandra in Wistar albino rats. Materials and Methods: It was a laboratory-based experimental study. Acetic acid-induced writhing test was used to determine the analgesic activity and the histamine-induced paw edema formation for anti-inflammatory activity of the extract. Twenty eight animals were divided into 7 groups each with 4 rats and two (2) sets of data were obtained from each animal per group as per guidance of the refinement rule of 3Rs. For analgesic activity, Group I was dosed with 1.0ml of normal saline (negative control), group II, III, IV, V, VI were dosed with 1000.0, 600.0, 150.0, 40.0 and 5.0 mg/kg bwt of extract respectively. Group VII was dosed 10.0 mg/kg bwt diclofenac sodium (Na) drug (positive control). For anti-inflammatory activity, Group I was dosed with 1.0ml of normal saline (negative control), Group II, III, IV, V, VI were dosed with 1000.0, 600.0, 300.0, 40.0 and 5.0 mg/kg bwt of extract respectively. Group VII was dosed 10.0 mg/kg bwt diclofenac Na. The percentage mean paw volume inhibition and percentage writhing protection were determined. Results: The mean percentage paw volume inhibitions were 29.1%, 74.2% and 32.3% at doses of 5.0, 40.0 mg/kg bwt and 10.0 mg/kg bwt diclofenac Na respectively. The mean percentage writhing protection was 8.9%, 10.4%, 78.5% and 98.7% at doses of 150.0, 600.0, 1000.0 mg/kg of extract and 10.0 mg/kg bwt diclofenac Na respectively. Conclusions: Total crude leaf extracts of P. dodecandra has analgesic and anti-inflammatory activity that may be attributed to the presence of polyphenolic, saponins and many others phytochemicals that are present in the plant.

EFFECTS OF PHYTOLACCA ACINOSA POLYSACCHARIDES I ON CYTOTOXICITY OF MACROPHAGES AND ITS PRODUCTION OF TUMOR NECROSIS FACTOR AND INTERLEUKIN 1

The in vivo effects of Phytolacca acinosa poly-saccharides I (PEP-I) on immunologic cytotoxicity of mouse peritoneal macrophages and its production of tumor necrosis factor (TNF) and interleukin 1 (IL-1) were studied. PEP-I 80 or 160 mg kg was given ip twice every 4 day. Both doses were found to have significant enhancing activity on macrophages cytotoxicity against S180 sarcoma cells and malignant transformed fibroblast L929 cells. Peritoneal activated macrophages were incubated with LPS for 2 and 24 hrs to induce TNF and IL-1, respectively. The TNF and IL-1 activities were tested from cytotoxicity against L929 cells in an absorbence assay of enzymatic reaction and proliferation of thymocytes co-stimulated assay separately. The optimal time for TNF production was found on day 8. Significant increases in TNF and IL-1 were observed. In comparison of the effect of PEP-I on TNF with that of known priming agent BCG, there was no difference between them, but PEP-I had a high effect on IL-1. These results suggest that cytotoxicity of macrophages primed by PEP-I is closely related to its TNF and IL-1 production.

中国大陆地区一种归化植物新记录——二十蕊商陆(Phytolacca icosandra L.)

商陆属(Phytolacca L.)主要分布于亚洲、非洲和美洲,全世界约有26种。中国本土分布4种,生长于沟谷、林下、林缘、路边等生境。该研究报道了中国大陆地区1种新归化植物:二十蕊商陆(P.icosandra L.)。该种心皮7—10枚,合生;花梗通常0.1—0.4(—1)cm,花被片5—6枚,不等大或等大,雄蕊14—20枚;果序直立,长20—30 cm,种子种脐一侧明显突出,与中国大陆地区记载的商陆属其他物种区别较明显而容易识别。该种原产于拉丁美洲,2012年归化于中国台湾,在欧洲、南非、澳大利亚、亚洲热带亚热带地区也有归化,因其具有较强环境适应性已成为部分归化地区的恶性杂草。文章介绍了二十蕊商陆的学名、文献引证、异名,然后结合野外观察和该种的凭证标本,对其特征进行了详细描述。通过该种与其相似种的形态学比较,文章编制了实用的中国商陆属植物检索表。此外,基于该种在归化地生长和分布情况的初步观察,文章还评估了该种的经济价值与入侵风险,认为该归化种具有较高入侵潜力。根据其生物学特性和其他地区的入侵表现,文章预测该种将来可能会扩散到中国更多省区。因此,文章提出了多条针对该种的归化管理对策。建议有关部门和人员应对其开展定期监测和相关研究,密切关注其归化种群的发展扩散趋势,防止其对中国生态系统造成危害。

商陆提取物药理作用研究进展

商陆是我国传统峻下逐水中药,收载于历版《中华人民共和国药典》,入药部位为根部,具有利尿泻下、抗炎、抗肿瘤、祛痰镇咳平喘等药理活性。临床多用于治疗肾炎、肝硬化腹水、乳腺增生、银屑病等疾病。通过对商陆提取物和化学成分药理作用进行总结,为今后商陆深入研究及合理开发利用提供支持和参考依据。

商陆(Phytolacca acinosa)微茎尖培养及多倍体诱导

本研究对商陆(Phytolacca acinosa)进行茎尖微茎尖培养与多倍体诱导研究。研究结果表明:适合商陆微茎尖生长的培养基为MS+6-BA 1.0 mg/L+IBA 0.02 mg/L;适合诱导商陆微茎尖生根的培养基为1/2MS+NAA 0.5 mg/L+IBA 1.0 mg/L,诱导率为100%。用0.3%的琼脂溶胶配制成秋水仙素浓度为0.2%的溶胶药剂连续处理微茎尖48 h,获得变异植株,表现为植株健壮,叶片皱缩增大,颜色加深,气孔数目增多、增大。压片法对变异植株根尖进行染色体鉴定,证明变异株为多倍体。

超积累植物垂序商陆(Phytolacca americana L.)吸收锰机制的初步探讨

垂序商陆是中国境内发现的锰超积累植物,了解其对锰的吸收特性及机制可为植物修复提供基础信息.通过人工培养液温室培养方法,探讨了低锰浓度处理下垂序商陆吸收累积锰的特性;采用同步辐射荧光技术(SRXRF)分析了锰在垂序商陆根横截面的分布特征;应用能量代谢抑制剂2,4-二硝基苯酚(DNP)和矾酸钠(Na3VO4)、钙离子通道抑制剂氯化镧(LaCl3)研究了垂序商陆对锰吸收,并探讨了其吸收机制.结果表明,低锰浓度处理下,锰超积累植物垂序商陆仍然具有较高的吸收和累积锰的能力,5μmol·L-1低锰浓度处理下垂序商陆根、茎、叶锰含量(以干重计)仍可高达402、208、601 mg·kg-1.锰在垂序商陆根横截面中柱含量最高,其次为表皮,皮层锰含量最少,锰从皮层进入到中柱是一个从低浓度到高浓度的过程,表明垂序商陆对锰的吸收可能存在主动运输过程.DNP和Na3VO4对垂序商陆锰吸收有一定的抑制作用,表明锰进入细胞是需要消耗能量的,进一步说明垂序商陆对锰的吸收存在主动吸收的过程.钙离子通道抑制剂LaCl3抑制垂序商陆根对锰的吸收高达30%,表明垂序商陆对锰的吸收与钙离子通道密切相关.

Effects of generalist herbivory on resistance and resource allocation by the invasive plant, Phytolacca americana

Successful invasions by exotic plants are often attributed to a loss of co-evolved specialists and a re-allocation of resources from defense to growth and reproduction. However, invasive plants are rarely completely released from insect herbivory because they are frequently attacked by generalists in their introduced ranges. The novel generalist community may also affect the invasive plant＇s defensive strategies and resource allocation. Here, we tested this hypothesis using American pokeweed （Phytolacca americana L.）, a species that has become invasive in China, which is native to North America. We examined resistance, tolerance, growth and reproduction of plant populations from both China and the USA when plants were exposed to natural generalist herbivores in China. We found that leaf damage was greater for invasive populations than for native populations, indicating that plants from invasive ranges had lower resistance to herbivory than those from native ranges. A regression of the percentage of leaf damage against mass showed that there was no significant difference in tolerance between invasive and native populations, even though the shoot, root, fruit and total mass were larger for invasive populations than for native populations. These results suggest that generalist herbivores are important drivers mediating the defensive strategies and resource allocation of the invasive American pokeweed.

Dispersal of invasive Phytolacca americana seeds by birds in an urban garden in China

Although seed dispersal is a key process determining the regeneration and spread of invasive plant populations,few studies have explicitly addressed the link between dispersal vector behavior and seedling recruitment to gain insight into the invasion process within an urban garden context.We evaluated the role of bird vectors in the dispersal of pokeweed(Phytolacca americana),a North American herb that is invasive in urban gardens in China.Fruiting P.americana attracted both generalist and specialist bird species that fed on and dispersed its seeds.The generalist species Pycnonotus sinensis and Urocissa erythrorhyncha were the most frequent dispersers.Seedling numbers of P.americana were strongly associated with the perching behavior of frugivorous birds.If newly recruited bird species use seedling-safe perching sites,the P.americana will regenerate faster,which would enhance its invasive potential.Based on our observations,we conclude that the 2 main bird vectors,P.sinensis and U.erythrorhyncha,provide potential effective dispersal agents for P.americana.Our results highlight the role of native birds in seed dispersal of invasive plants in urban gardens.