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.

镉胁迫对商陆生理指标的影响

[目的]研究不同浓度的镉胁迫对商陆生理指标的变化。[方法]以商陆(Phytolacca acinosa Roxb.)为材料,通过温室盆栽的方式,研究其在0、10、25、50和100 mg/kg镉处理下部分生理指标的变化。[结果]在浓度小于10 mg/kg时,商陆叶片中叶绿素a含量和可溶性蛋白的含量、CAT和POD活性随浓度的增加而增大;而当浓度大于10 mg/kg时,商陆叶片中叶绿素a含量和可溶性蛋白的含量、CAT和POD活性随浓度的增加而降低;商陆叶片中MDA含量随镉浓度的升高而增大;叶绿素b含量、可溶性糖的含量则随着镉浓度的升高而下降。[结论]商陆能够通过调节体内抗氧化酶系统的活性,并保持其较高的水平以抵御镉胁迫造成的危害,从而保护了细胞膜的稳定性和完整性,显示了其超积累植物的特性。

Mn超富集植物商陆的多倍体诱导与修复性能

以Mn超富集植物商陆(Phytolacca acinosa Roxb)幼苗为试验材料,研究了不同秋水仙素浓度和处理时间对商陆多倍体诱导的影响,并对诱导的多倍体与二倍体进行了形态、染色体、生物量和修复性能的鉴定比较。结果表明,0.5 g/L的秋水仙素处理24 h诱导效果最好,变异率达到46.7%;多倍体植株的叶宽、叶厚、花直径和气孔面积等形态参数均显著大于二倍体;根尖细胞的压片镜检表明,变异材料的染色体数(2n=4x=72)为二倍体对照(2n=2x=36)的两倍;不同Mn浓度处理时多倍体商陆根、茎和叶的生物量(干重)均比二倍体增加,且多倍体平均单株Mn富集量达到14.85 mg,远高于二倍体的7.73 mg。

几种孕妇禁忌中药对早孕绒毛的组织化学及超微结构的影响

本文从形态学及组织化学角度,观察了孕妇禁忌中药商陆、附子及蒲黄对器官培养的早孕绒毛的影响。结果表明:商陆对绒毛的损伤最为严重,光镜观察可见绒毛萎缩变小,滋养层与间质分离,细胞核固缩、溶解、碎裂;胞质减少且嗜酸性增强;电镜下可见,核染色质及核仁溶解,核膜皱缩不规则;各种细胞器损伤较为严重,形态结构难以分辨;胞浆中可见大量的自噬泡和髓样体;组织化学显示,ACP反应增强;ALP反应减弱,3β-HSD、SDH及TPPase反应极弱,甚至阴性。附子对绒毛也有一定程度的损伤,但较商陆为轻。蒲黄对绒毛未见明显损害。

Protective Effect of phytolacca Acinosa Polysaccharides Ⅰ(PAP-Ⅰ)on Hematopoiesis in Cyclophosphamide-treated and 60￣Co-irradiated Mice

The protective effect of a kind of purified polysaccharides extracted from Radix of Phytolacca acinosa Roxb,with a molecular weight of 10 KDa,on hematopoiesis was investigated.Average survival time of mice treated with cyclophosphamide (CY) 300 mg/kg once alone was 13.3 ± 7.2d(n=7) however,average survival time of mice treated with CY 300 mg/kg in com-bination with PAP-1 10 mg/kg,3 times/wk was 36.7± 16.4d(n=7,P<0.01).PAP-1,ip had benefi-cial effect on the recovery of the CY induced decrease of peripheral leukocyte number,and the nu-cleated bone marrow cell(BMC)number and[3￣H]TdR uptaken by BMC induced by rmGM-CSF in S180 bearing mice treated with CY,In mice,after the first ip treatment with CY 100 mg/kg on d7,the peripheral leukocyte number decreased on d9 and recovered to normal level about d13 to d15. Such recovery was accelerated by administrating PAP-1,10mg/kg, 3 times/wk.A significant in-crease of the activity to form colony in spleen(colony-forming unit in spleen, CFU-S_8, CUF-S12) in mice irradiated with 550 rad 6O￣Co γ-rays and an enhancement of proliferative response of BMC to rmGM-CSF treated with PAP-1,10mg/kg,3 times/wk, ip were observed.After PAP-1,10 mg/kg,ip once,a significant increase in the number of peripheral blood leukocytes and a rise in the serum of colony stimulating factor(CSF) were also confirmed.The types of CSF in serum were M-CSF and other hematopoietic growth factors,which were confirmed by using McAb of IL-3, GM-CSF and PcAb of M-CSF. These beneficial effects of PAP-1 on hematopoiesis may be related to its activityinducing CSFs and other hematopoietic growth factors and warrant further evaluation of its use-fulness.

Antifeedant Activity of Three Plant Extracts Including P. acinosa against Pieris rapae

[Objective] The research aimed to study antifeedant activity of Phytolacca acinosa Roxb., Setaria viridis （L.） Beauv and Viola yedoensis Makino extracts against Pieris rapae. [Method] Activity material was extracted from S. viridis （L.）, P. acinosa and V. yedoensis using acetone cold soak method, and non-selective antifeedant activity of extracts to Pieris rapae larva was determined by using lobular plate addition method. [Result] The results showed that the acetone leaching agent of P. acinosa had most obvious antifeedant effects on Pieris rapae. The antifeedant rate were 74.53% and 82.34% at 24 and 48 h respectively. With the concentration increasing, the antifeedant effect of P. acinosa extracts increased. The antifeedant rate of 0.050 g/ml treatment was the highest, being 74.53% and 82.34% at 24 and 48 h. [Conclusion] P. acinosa could be studied and utilized as potential botanical insecticide.

不同干燥方法对商陆总皂苷和皂苷甲含量的影响

利用硫酸-香草醛比色法测定商陆(Phytolacca americana)总皂苷含量,采用HPLC-UV法测定商陆皂苷甲的含量。结果表明,商陆总皂苷和皂苷甲的含量比较,阴干商陆明显低于其他组,晒干商陆次之,烘干品含量较高;其中低温55℃烘干品的商陆总皂苷含量最高,80℃烘干的商陆皂苷甲含量高于其他组。商陆产地加工中,应及时干燥,避免成分流失,规模化烘干优于传统晒干,采用低温烘干为宜。

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.

HS-SPME-GC-MS分析商陆及其炮制品挥发性成分

利用顶空固相微萃取技术(HS-SPME)与气相色谱-质谱联用技术(GC-MS)对比分析商陆(Phyto鄄lacca acinosa Roxb.)不同炮制品挥发性成分,并通过面积归一化法计算每种成分的相对百分含量。结果表明,从商陆生品中检测出39个峰,鉴定出29种成分,占挥发性成分的76.52%;从醋商陆中检测出33个峰,鉴定出27种成分,占挥发性成分的68.78%。共有挥发性成分有14种,与商陆生品相比,醋商陆减少15种成分,新增13种成分。商陆不同炮制品中挥发性成分的组成和含量均发生了变化。

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.

垂序商陆对土壤中镉的累积动力学及施肥影响

设置不同浓度处理与施肥处理的温室盆栽实验,研究垂序商陆(Phytolacca americana)对土壤中Cd的累积动力学,以及硫酸铵〔(NH 4)2 SO 4〕和尿素(CH 4 N 2 O)作为氮肥、氯化钾(KCl)作为钾肥对垂序商陆生长和吸收累积Cd的影响。结果表明,在土壤Cd含量为10 mg·kg^-1时,垂序商陆表现出最高的累积系数(BCF,10.37)和迁移速率(TF,2.78),在15~30 d时累积速率最高(21.1μg·d^-1),吸收速率常数也较高〔(0.21±0.01)d^-1〕;垂序商陆对Cd的吸收速率常数随着暴露时间的增加而逐渐降低,15、30、60和90 d时分别为(0.35±0.02)、(0.21±0.01)、(0.11±0.01)和(0.09±0.01)d^-1;分别使用(NH 4)2 SO 4和CH 4 N 2 O作为氮肥,当N含量为400 mg·kg^-1时,垂序商陆地上部Cd含量分别增加183.6%和142.3%,生物量分别提高165.3%和150.4%,Cd累积量分别增加478.7%和337.3%;使用KCl作为钾肥,当K含量为600 mg·kg^-1时,垂序商陆地上部Cd含量提高264.4%,但生物量仅提高1.4%,钾肥处理组垂序商陆Cd累积量低于氮肥处理组。

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.

Influence of SLYWG on malignant tumor ascites

Objective To search the effect of Chinese traditional medicine "Shang Lu Yu Wang Gao"(SLYWG)on the ascites induced by tumor,and its mechanism.Methods The tumor ascites model and diuretic experiments were introduced to evaluate the effect of SLYWG.Physical characteristics,the tumor cell counting,volume of the ascites,protein content in ascites,the characters of ascites,the life duration of S180 tumor bearing animals as the indexes of evaluation.the diurtic experiments were performed on rats nad rabbits,the osmotic pressure,K+,Na+,Cl-,Ca2+ and pH in urine were determined.Results The inhibitions to ascites of SLYWG were displayed in three dosage(30 g·kg-1,15 g·kg-1 and 7.5 g·kg-1).Ascites caused by tumor was significantly inhibited by the local administration of SLYWG.The increase of mice ascites was slow down,the content of ALB and the TP in ascites were decreased,the surviving time of mice was extended.SLYWG had remarkable diuresis effect on the rats and rabbits,it could reduce the osmotic pressure of urine,decrease the exclude of K+ but had no effect on the Na+,Cl-,Ca2+ and pH in urine.Conclusions Tumor ascites was significantly inhibited by the ventral administration of SLYWG.SLYWG had diuretic effect in rats and rabbits,it reduced the osmotic pressure of urine,decrease the exclude of K+ but had no effect on Na+,Cl-,Ca2+ and pH of urine.

Cloning and Sequencing of the Pokeweed Antiviral Protein Gene and Its Expression in E. coli

The total RNA was isolated from pokeweed (Phytolacca americana) leaves using the method of guanidine isothiocyanite and used as a template to amplify the deleted mutant pokeweed antiviral protein (PAP) gene by RT-PCR and then the gene was cloned into the pGEMR-T vector. The sequencing results showed that the PAP gene consisted of 711nt, which was 99.6% identical to the PAP gene reported by Lin et al (1991). The IPTG-inducible expression vector containing the PAP gene was constructed and transferred into the E. coli strain BL21 (DE3)-plysS. A specific protein was produced after induction with 0.4m mol/L IPTG and its molecular weight was 26ku. The results of the double diffusion on the agar plate and the western blotting test showed that the protein produced in E. coli was highly identical with the PAP extracted by a Frenchman from French pokeweed leaves. These revealed that PAP gene was actually achieved and exactly expressed in E. coli.

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).

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.

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.

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.

Medicinal properties and anti-inflammatory components of Phytolacca(Shanglu)

Phytolacca(Shanglu)is a well-known traditional herbal medi-cine that has been used for thousands of years in China.Phytolacca is also used worldwide in different traditional phyto-medicines and homeopathic preparations.The present work re-viewed advances in the traditional uses,plant origin,chemical constituents,pharmacology,and medicinal properties of Phytolacca.Phytolacca is usually made from the roots of Phytolacca acinosa and P.esculenta,but the invasive plant P.americana(American pokeweed)is also widely used.Different types of medicinal products made with Phytolacca are available,as well as various types of Phytolacca extracts,showing a range of pharmacological activities including antioxidant,anti-inflammat-ory,anti-parasitic,antifungal,anticancer,and insecticidal effects.The marked anti-inflammatory activity of Phytolacca extracts supports its use in treating diseases such as arthritis,nephritis,and rheumatism,as well as in combatting cancer.Several bioact-ive natural products have been identified from Phytolacca,in-cluding glycosylated saponins such as esculentosides and phytolaccosides,as well as a few flavones(cochliophilin A)and phytosterols(α-spinasterol),which contribute to Phytolacca’s anti-inflammatory and/or anticancer effects.A quality evaluation of Phytolacca-based extracts and products is highly recommen-ded;nevertheless,the use of Phytolacca can be encouraged to help regulate cytokine production and combat inflammatory dis-eases.