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.

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 。

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.

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.

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.

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。

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。

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.

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。

探究不同浓度商陆提取液对常见三种霉菌的抑制效果

以安徽菌启健公司的商陆为试验原材料,采用水提法提取商陆浸提液。用商陆水提液开展对常见霉菌抑制作用的实验。实验表明:在10~100 g/L浓度范围内,哈茨木霉的菌落直径由17.27 mm降至11.14 mm;层出镰孢菌的菌落直径由17.07 mm降至11.07 mm;茄腐镰刀菌的菌落直径由16.17 mm降至10.23 mm,其最佳抑制浓度为100 g/L。实验结果发现商陆水提液对哈茨木霉菌、层出镰孢菌、茄腐镰刀菌三种菌有较好的抑菌效果,其抑菌率分别为53.43%(哈茨木霉菌)、53.25%(层出镰孢菌)、59.37%(茄腐镰刀菌)。实验结果表明:一定浓度的商陆水煮提取液能有效抑制哈茨木霉菌、层出镰孢菌、茄腐镰刀菌的生长,在防霉实践中具有潜在应用前景。

螯合剂和有机酸对商陆修复镉砷污染农田的影响

基于利用植物提高Cd和As复合污染农田修复效率的目标,通过大田试验方法探究了乙二胺四乙酸(EDTA)、皂素(SAP)、柠檬酸(CA)和苹果酸(MA)对商陆萃取农田土壤Cd和As的影响。结果表明,施用4种不同螯合剂和有机酸对商陆的生长没有产生不利影响,且可以显著影响商陆对土壤Cd和As的吸收积累。与对照组相比,EDTA、SAP、CA和MA处理60 d后导致商陆叶片Cd含量分别提高116.4%、55.0%、81.5%和109.9%,商陆地上部Cd积累量提高40.4%~106.0%,商陆地上部As积累量提高15.8%~55.1%。商陆各器官Cd和As的富集系数和转运系数也受螯合剂和有机酸施用的影响。施用EDTA、SAP、CA和MA均对商陆根际土壤Cd和As含量产生显著影响,SAP处理对降低商陆根际土壤Cd含量的影响最显著,而MA处理对降低商陆根际土壤As含量的影响最显著。研究表明,EDTA、SAP、CA和MA均可提高商陆萃取农田土壤Cd和As的效率,尤其是EDTA和MA的效果更好。

中国首次发现的锰超积累植物——商陆

自 2 0 0 0年以来 ,对位于湖南省湘潭锰矿污染区的植物和土壤进行了一系列的野外调查 ,以着力寻找锰的超积累植物。结果表明 ,商陆科植物商陆对锰具有明显的富积特性 ,叶片内锰含量最高达 1 92 99mg/ kg。这一发现填补了我国锰超积累植物的空白 。

锰在商陆叶片的细胞分布及化学形态分析

商陆(Phytolacca acinosa Roxb.)是在中国境内首次发现的锰超积累植物,叶片是商陆累积锰的主要器官。本文主要采用化学分析、差速离心及色谱技术对商陆叶片锰的细胞分布和化学形态进行了研究并探讨了商陆超累积和解毒锰的机制。细胞分配的结果表明,锰主要分布在商陆叶片的细胞质可溶性部分(含液泡),含量高达74%-82%,细胞壁结合的锰占总锰的15%-20%,细胞器锰含量最少,仅为3%-6%。不同提取剂的实验结果表明,商陆叶片中的Mn均以水提取态为主,为总Mn含量的83%-91%;不同锰浓度处理下,商陆叶片草酸含量高于500μmol·g-1(干重),达到干重的9%-11%,但不同锰浓度处理下草酸的含量并无明显变化趋势,表明高含量的草酸的累积是商陆本身具有的特性,而非外界锰的诱导产生。

商陆多糖Ⅰ对荷S180小鼠的抑癌,增强免疫和造血保护作用

商陆多糖Ⅰ(PEP Ⅰ)10,20mg·kg^1 ip能显著抑制S180生长,最大抑瘤率可达51.7％,Cy 10mg·kg^1 ip.抑瘤率为55.1％,两者合用抑瘤率为68.6％,PEP-Ⅰ治疗能显著促进脾脏增生,但不影响体重,PEP-Ⅰ治疗后,小鼠的T淋巴细胞转化及白介素2(IL-2)产生能力显著高于对照组,计数外周白细胞,骨髓有核细胞并用[~3H]TdR参入法观察rmGM-CSF刺激骨髓细胞增殖.结果表明PEP-Ⅰ与Cy合用治疗至d 11的外周白细胞数,骨髓有核细胞数和骨髓细胞的增殖能力显著高于单用Cy组,以上实验结果说明EPP-Ⅰ可能通过增强T淋巴细胞功能来抑制移植性肿瘤生长,并具有保护造血功能的作用。

商陆多糖I对小鼠淋巴细胞DNA多聚酶α活性的影响

本实验在建立ＤＮＡ多聚酶α活性测定方法的基础上观察了商陆多糖Ｉ（ＰＡＰ－Ｉ）对小鼠淋巴细胞增殖及淋巴细胞ＤＮＡ多聚酶α活性的影响。采用［３Ｈ］－ＴｄＲ掺入法和模板活化法（［３Ｈ］－ｄＴＴＰ掺入法）分别检测ＰＡＰ－Ｉ给药组及对照组小鼠淋巴细胞增殖能力和淋巴细胞ＤＮＡ多聚酶α活性。体外实验发现ＰＡＰ－Ｉ显著增强ＣｏｎＡ诱导的小鼠脾淋巴细胞增殖及其ＤＮＡ多聚酶α的活性；小鼠腹腔注射ＰＡＰ－Ｉ（每周１次）对脾淋巴细胞ＤＮＡ多聚酶α活性基本上无影响，但加入ＣｏｎＡ（５μｇ／ｍｌ）刺激后，ＰＡＰ－Ｉ１０ｍｇ／ｋｇ剂量组ＤＮＡ多聚酶α活性显著增强。结果提示ＰＡＰ－Ｉ增强ＤＮＡ多聚酶α活性可能是促进脾淋巴细胞增殖、增强免疫功能的机制之一。

中国商陆抗病毒蛋白基因的克隆及其转化辣椒

根据美洲商陆抗病毒蛋白基因序列设计引物,从中国商陆(Phytolacca acinosa)叶片基因组DNA扩增克隆缺失无毒型中国商陆抗病毒蛋白基因(PacPAP1),构建该基因植物表达载体,采用农杆菌介导的叶盘法转化辣椒,对转基因植株进行分子检测,TMV及CMV人工接种鉴定,25d后统计发病情况。结果表明:克隆得到缺失无毒型PAP基因PacPAP1,大小为714bp,与美洲商陆抗病毒蛋白基因(Pam-PAP)的同源性为99.7%;得到了PacPAP1整合入辣椒基因组中的阳性植株;接种TMV、CMV的阴性对照植株都明显发病,转PacPAP1植株未出现症状,说明转基因辣椒植株可获得抗TMV和CMV材料。

商陆不同极性、根和茎提取物的抑菌性能分析

为探究商陆不同极性、部位提取物的抑菌性能,本实验分别从商陆根和茎中提取石油醚相、乙酸乙酯相、正丁醇相和水相4种不同极性的提取物,再采用滤纸片法测量提取物对大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphyloccocus aureus)、巨大芽孢杆菌(Bacillus megaterium)和副溶血弧菌(Vibrio parahaemolyt-icus)4种细菌的抑制性能。结果表明,部分商陆的提取物有一定的抑菌活性,如根正丁醇相对巨大芽孢杆菌和副溶血弧菌,茎水相对副溶血弧菌的抑菌圈在10mm以上,但总体上商陆的提取物的抑菌性能远不及头孢氨苄好。实验还发现商陆抑菌活性最强的物质大都存在于根中,且抑菌活性最强的物质大都存在于水和正丁醇这种极性较高的溶剂的提取物中,不同提取物对不同细菌的抑菌活性情况不一致。因此本研究结果可以为商陆提取物抑菌性能的进一步探索提供参考。

商陆对镉锌铜胁迫的生理响应与金属积累特性

以水培方法研究了商陆(Phytolacca acinosa)在不同浓度镉、锌和铜处理下的叶绿素含量、超氧化物歧化酶(SOD)活性、生物量和组织重金属含量。结果表明,随着处理浓度增加,叶绿素含量、SOD活性和生物量呈先增加后降低趋势。组织重金属含量在镉、锌、铜为最高浓度处理时达到最高值。根重金属含量高于茎和叶。叶片中重金属含量的最大值分别为镉100.6mg/kg、锌2753mg/kg、铜36.9mg/kg。商陆对锌的蓄积能力接近于超积累植物东南景天,是一种在锰锌复合污染土壤植物修复中具有较大应用潜力的种质资源。

药用植物商陆毛状根培养系统的建立

目的:建立药用植物商陆(Phytolacca acinosa Roxb.)的毛状根离体培养系统。方法:分别用发根农杆菌A4、R1600、C58C13个菌株感染商陆的子叶外植体,获得毛状根,并筛选了优质株系;考察了影响毛状根生长的因素;测定了毛状根的生长曲线;利用PCR和高压纸电泳法对毛状根进行tDNA转化的检测。结果:利用发根农杆菌A4、R1600、C58C13种菌株成功地从商陆中诱导出毛状根;以水解酪蛋白为氮源、以麦芽糖为碳源最有利于商陆毛状根的生长;毛状根的生长曲线呈“S”形,24 d时质量达到最大(增长18倍);PCR及高压纸电泳检测结果表明Ri质粒的tDNA已整合进毛状根中。结论:成功建立了商陆毛状根离体培养系统,为进一步进行药用成分的工业化生产和引入外源基因改良性状奠定了基础。

商陆对镉的富集特征

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。对位于湖南省株洲冶炼厂厂区的植物进行了一系列的野外调查,以着力寻找镉的超积累植物。结果表明,商陆科Phytolaccaceae植物商陆Phytolacca acinosa对镉具有明显的富积特性,商陆在镉质量分数最高达106.0 mg.kg-1厂区土壤中依然生长良好,茎叶中镉质量分数高达63.8 mg.kg-1。盆栽条件下,商陆对生长介质中的镉具有很强的耐性和累积能力。当生长介质中镉质量分数为5.0 mg.kg-1时,其生物量与对照相比有所增加。随着生长介质中镉质量分数的升高,虽然其生物量与对照相比有所降低,但植株仍生长良好。其地上和地下部分镉的质量分数逐渐增加,生物富集系数则逐渐降低。当土壤中镉的质量分数达到200.0 mg.kg-1时,商陆仍能完成整个生命周期。此外,还利用吸收量系数对商陆对镉的去除能力和富集特征进行了评价,进一步验证了商陆的镉超积累特性。