Increased production of plumbagin in Plumbago indica root cultures by gamma ray irradiation

Abstract

Context: Plumbagin is a major active constituent of Plumbago indica L. (Plumbaginaceae). It possesses various pharmacological activities that have been shown to assist in the treatment of various diseases.

Objectives: This work is focused on increasing the production of plumbagin in P. indica root cultures using low doses of gamma ray irradiation as an elicitor.

Materials and methods: The effect of low doses of gamma ray irradiation (0, 5, 10, 15, 20, 25?Gy) and ages of the root cultures (0, 5, 10, 15, 20 days) for elicitation of plumbagin production was determined. The stability of the elicited root cultures to produce plumbagin was also determined during three cycles of subculture.

Results and discussion: Treatment of the root cultures with a low dose of gamma ray at 20?Gy gave the highest level of plumbagin production (1.04?mg/g DW) when compared to all other treated groups. The appropriate age of the root cultures for maximum production of plumbagin was found to be 10 days. However, treatment of 5-day-old root cultures resulted in a significant increase of dried root biomass that also had a high plumbagin production. Based on the total biomass per culture flask, the amounts of plumbagin produced by the 5- and 10-day-old treated roots were 0.59 and 0.37?mg/250?mL flask, respectively, which were 4.2- and 2.6-fold higher than the level in the control. Subculturing the root cultures until the third generation still showed an increase in plumbagin production without any effects on their growth.     

RP-HPLC测定民族药材白花丹不同药用部位中白花丹醌的含量

目的：建立白花丹药材中白花丹醌的含量测定方法并考察白花丹不同药用部位中的含量。方法：色谱条件：Kromasil C₁₈柱(4.6 mm×200 mm,5 μm),柱温30 ℃。流动相甲醇-水（65∶35）,检测波长213 nm,流速1 mL·min^-1。结果：白花丹醌在0.020 8～0.104 μg,峰面积与进样量具有良好的线性关系,回归方程Y=-14.29+9 138.9 X,r= 0.999 9,平均回收率为98.7%。白花丹各部位中白花丹醌的含量分别为：根0.394%,茎0.050%,叶0.031%。结论：本法简便、准确、重复性好,可用于控制白花丹药材质量。     

白花丹化学成分的研究

目的:研究白花丹Plumbago zeylanicaLinn.地上部分的化学成分。方法:对白花丹95%乙醇提取物的乙酸乙酯部分进行色谱分离,通过理化性质和波谱分析鉴定化合物的结构。结果:从白花丹中分离得到9个化合物,分别鉴定为白花丹醌(Ⅰ),isoshinanolone(Ⅱ),白花丹酸(Ⅲ),β-谷甾醇(Ⅳ),对羟基苯甲醛(Ⅴ),反式桂皮酸(Ⅵ),香兰子酸(Ⅶ),2,5-二甲基-7-羟基-色原酮(Ⅷ),3-吲哚甲醛(Ⅸ)。结论:化合物Ⅴ,Ⅶ,Ⅷ和Ⅸ为首次从白花丹属植物中分离得到。     

多枝雾水葛不同提取部位的抗炎镇痛作用研究

目的 研究多枝雾水葛不同提取物的抗炎镇痛作用。方法 采用二甲苯致小鼠耳肿胀、醋酸致小鼠扭体反应和小鼠热板法观察多枝雾水葛灌胃、外搽使用以及不同提取部位的抗炎、镇痛作用。结果 多枝雾水葛外搽使用的抗炎镇痛效果不明显,灌胃给药具有较强的抗炎镇痛作用,能显著减少醋酸所致的小鼠扭体次数和抑制二甲苯所致的小鼠耳廓肿胀,且其水提物比醇提物的作用更强。对多枝雾水葛水提取物的活性部位研究表明,多枝雾水葛的氯仿部位和水部位都具有较好的镇痛效果,抗炎的有效部位则是氯仿部位和正丁醇部位。结论 多枝雾水葛水提取物灌胃给药具有抗炎镇痛作用,其有效部位是氯仿部位、正丁醇部位和水提取部位。     

Plumbago auriculata leaf extract-mediated AgNPs and its activities as antioxidant,anti-TB and dye degrading agents

In the present work, silver nanoparticles have been biosynthesized by utilizing the alcoholic extract of Plumbago auriculata. The optimization of reaction conditions was carried out by monitoring the reactions with the help of UV–Visible absorption spectroscopy. The characterization of AgNP was carried out by infrared spectroscopy, transmission electron microscopy and X-Ray diffraction (XRD) studies. The biogenic AgNPs were tested against Mycobacterium tuberculosis using Microplate Almar Blue assay (MABA) and their antioxidant activity was also evaluated. The silver nanoparticles were also assessed for their reducing activity against organic dyes. The AgNPs were spherical in shape with size ranging from 15 to 45 nm with face centered cubic geometry as revealed by XRD analysis. The AgNPs possessed good antitubercular activity with MIC value of 1.6 μg/ml and these also exhibited promising antioxidant activity with IC₅₀ value of 28.2. Furthermore, AgNPs also reduced congo red within 2 h and malachite green was degraded within 40 min. The present work demonstrated the utilization of P. auriculata for biosynthesis of AgNP which could be a potential candidate for antitubercular drug development and it could also be used as an antioxidant agent. The application of AgNP in reducing agent can be further extended and evaluated for purification of effluent water from textile industries.     

ICP-OES法测定不同栽培条件下白花丹茎中的无机元素

目的 测定不同栽培条件下白花丹茎样品中无机元素的含量.方法 采用微波肖解和电感耦合等离子体发射光谱法(ICP-OES)同时测定白花丹茎中的无机元素.结果 栽培条件对白花丹茎中无机元素的含量影响较大,且大都降低其无机元素的含量;栽培能大幅降低白花丹茎中As、Pb、Cd、Cr等重金属元素的总含量.结论 从减少重金属元素含量的角度分析,白花丹的栽培条件为:繁殖选用种子苗或扦插苗、植被条件选用全光照、施肥选用钾肥、土壤条件选择黏土.     

Genotoxicity and Interference with Cell Cycle Activities by an Ethanolic Extract from Thai Plumbago indica Roots in Human Lymphocytes in vitro

In Thai traditional medicine, Plumbago indica or Jetamul-Pleung-Dang in Thai is known to have healthbenefit especially for anti-inflammatory, antibacterial, and antitumor activities. However, the mechanisms ofits action are still uncertain. One of which might be genotoxic effects. In the present study, we investigated thegenotoxicity of an ethanolic extract of Plumbago indica root (EEPIR) by sister chromatid exchange (SCE) assayin human lymphocytes. Results have shown that all treatments with EEPIR (12.5-100 μg/ml) could induce cellcycle delay as shown by significant increase in the number of metaphase cells in the first cell cycle but neither inthe second nor the third cell cycle. Only at concentrations of 25, 50, and 100 μg/ml were SCE levels significantlyincreased above that of the control (p<0.05) . EEPIR at a concentration of 500 μg/ml induced cell death as fewmitotic cells were shown. Accordingly, EEPIR (25-100 μg/ml) is genotoxic in human lymphocytes and cytotoxic atconcentrations of ≥500 μg/ml in vitro. Therefore, these activities of the EEPIR could serve its potential therapeuticeffects, especially as an anticancer agent. Further study of EEPIR in vivo is now needed to support this in vitroevidence.     

多枝雾水葛凝胶膏剂抗皮肤感染的药效学研究

目的：研究多枝雾水葛凝胶膏剂（Pouzolzia zeylanica var.microphylla gel,PZG）对皮肤感染性疾病的治疗作用。方法：建立金黄色葡萄球菌诱导的小鼠皮下脓肿模型和大鼠皮肤创伤性感染溃疡模型,通过检测体质量、皮肤脓肿体积或溃疡面积愈合率、皮肤细菌逆培养和局部皮肤病理形态、羟脯氨酸、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）和白介素-10（interleukin-10,IL-10）含量等指标,考察PZG对感染性皮下脓肿和皮肤溃疡的治疗作用。结果：PZG能显著缩小皮下脓肿体积,提高皮肤溃疡面积愈合率,且有一定的量效关系;PZG可抑制皮肤感染部位的细菌生长,改善局部皮肤病理形态;PZG可降低溃疡部位TNF-α和IL-10水平,增加组织中羟脯氨酸的含量,促进溃疡愈合。结论：中药有效部位制剂PZG具有良好的抗皮肤感染作用,机制可能与其综合的抗菌、抗炎作用有密切关系。     

Protection by Black Tea Extract Against Chromosome Damage Induced by Two Heavy Metals in Mice

The efficacy of infusion of black tea leaf, Camellia sinensis (Linn.) O.Kuntze (Theaceae), in reducing the cytotoxic effects of two heavy metal salts, chromium and arsenic, was tested in bone marrow cells of mice following dietary administration. Mice were given black tea infusion by gavage twice daily, in concentrations simulating human consumption, for 6 days. On day 7 after treatment with tea, separate sets of mice were given single doses of potassium dichromate and sodium arsenite, and then killed after 24 h. Control sets were treated with potassium dichromate or sodium arsenite separately and also observed after 24 hours. The concentration of each salt corresponded to 1/10th of its LD 50 value. Chromosomes were studied from bone marrow cells following the usual colchicine-hypotonic-fixation-air-drying Giemsa schedule. Both metallic salts were highly clastogenic when given alone, inducing a high frequency of chromosomal aberrations as compared with d istilled water. Tea alone, given twice daily for 6 days, was not clastogenic. When mice, given tea twice daily for previous 6 days, were treated with either of the two salts on day 7, the degree of chromosome damage induced was reduced significantly as compared with the salts given alone. This reduction was more significant for sodium arsenite as compared with potassium dichromate. Such protection against arsenic cytotoxicity by prolonged dietary administration of black tea infusion is of importance in view of the widespread exposure of human populations to arsenic damage through drinking water from tubewells in West Bengal, India and adjoining Bangladesh.     

Norlignans from Pouzolzia zeylanica var. microphylla and their nitric oxide inhibitory activity

Five new compounds, pouzolignan F [4-hydroxy-3-(3,5-dihydroxyphenyl)-2-[bis(4-hydroxy-3-methoxyphenyl)methyl]butyl acetate] (1), pouzolignan G [4-hydroxy-3-(3,5-dihydroxyphenyl)-2-[(4-hydroxy-3,5-dimethoxyphenyl)(4-hydroxy-3-methoxyphenyl)methyl]butyl acetate] (2), pouzolignan H [1,4-dihydroxy-3-(3,5-dihydroxyphenyl)-2-[bis(4-hydroxy-3-methoxyphenyl)methyl]butane] (3), pouzolignan I [1,4-dihydroxy-3-(3,5-dihydroxyphenyl)-2-[(4-hydroxy-3,5-dime thoxyphenyl)-(4-hydroxy-3-methoxyphenyl)methyl]butane] (4), and pouzolignan J [1,4-dihydroxy-3-(3,5-dihydroxyphenyl) -2-[(3,4,5-trimethoxyphenyl)(4-hydroxy-3-methoxyphenyl)methyl]butane] (5), along with two known compounds, indolyl-3-carboxylic acid (6) and uracil (7), were isolated from the aerial parts of Pouzolzia zeylanica (L.) Benn. var. microphylla (Wedd.) W.T.Wang. The structures of these compounds were characterized based on spectroscopic methods, including IR, NMR (¹H–¹H COSY, HSQC, HMBC, and NOESY), and HR-ESI/TOF-MS experiments. All the new norlignans were assayed for inhibitory activity against lipopolysaccharide (LPS)-induced nitric oxide (NO) production in mouse peritoneal macrophages.