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.     

Radiosensitizing effects of neem (Azadirachta indica) oil.

Radiosensitization by neem oil was studied using Balbc/3T3 cells and SCID cells. Neem oil enhanced the radiosensitivity of the cells when applied both during and after x-irradiation under aerobic conditions. Neem oil completely inhibited the repair of sublethal damage and potentially lethal damage repair in Balbc/3T3 cells. The cytofluorimeter data show that neem oil treatment before and after x-irradiation reduced the G(2) + M phase, thus inhibiting the expression of the radiation induced arrest of cells in the G(2) phase of the cell cycle. However, SCIK cells (derived from the SCID mouse), deficient in DSB repair, treated with neem oil did not show any enhancement in the radiosensitivity. There was no effect of neem oil on SLD repair or its inhibition in SCIK cells. These results suggest that neem oil enhanced the radiosensitivity of cells by interacting with residual damage after x-irradiation, thereby converting the sublethal damage or potentially lethal damage into lethal damage, inhibiting the double-strand break repair or reducing the G(2) phase of the cell cycle.     

Ultrastructural changes in Leydig cells and cauda epididymal spermatozoa induced by Azadirachta indica leaves in albino rats

The effects of Azadirachta indica leaves (500 mg/kg body weight, orally/day), testosterone (0.25 mg/kg body weight/day, intramuscularly) and a combination of these two were investigated as to whether Azadirachta indica leaves affect the morphology of Leydig cells and epididymal sperm and their organelles at the ultrastructural level. Azadirachta indica treated rats showed atrophic seminiferous tubules with widening intercellular spaces. Leydig cells exhibited characteristics of degeneration such as indented nuclei; the commencement of degeneration was evident from less chromatin; the reduced amount of a marked decrease in organelle content and scarcity of other cell types of the interstitium in highly vacuolated cytoplasm, which were seem from both light and electron microscopic observations. The number of Leydig cells and their nuclear diameter were reduced significantly. Pathological changes in the spermatozoa of the cauda epididymis were observed and the spermatozoa retained cytoplasmic droplets. It is suggested that regression of Leydig cells and the absence of interstitium cell types indicates an androgen deficiency, which affects the spermatozoa in the epididymis by disturbing the internal epididymal milieu. The antiandrogenic and antispermatogenic properties of Azadirachta indica leaves appear to lead to a reduced fertilizing ability of the sperm.     

不同产地了哥王药材中总黄酮的含量测定

目的：对不同产地了哥王药材中总黄酮的含量测定并进行比较。方法：样品制备后加5%亚硝酸钠溶液1mL,放置6min后,加10%硝酸铝溶液1mL,放置6min后,加4%氢氧化钠溶液10mL,15min后,甲醇定容至刻度线,摇匀,在波长λ=510nm下测定吸光度,以芦丁做对照。结果：江西、广西、湖南3个产地的了哥王药材中的总黄酮平均含量分别为2.19%、2.52%、1.72%,广西产4批药材中含量相差较小。结论：可见不同产地,因不同的生长环境、温度、湿度等因素对了哥王中总黄酮的含量影响较大。     

Azadirachta indica modulates carcinogen biotransformation and reduced glutathione at peri-initiation phase of benzo(a)pyrene induced murine forestomach tumorigenesis

The present study evaluated the effects of aqueous Azadirachta indica leaf extract (AAILE) on the activities of certain phase I (cytochrome P450, cytochrome b(5) and aryl hydrocarbon hydroxylase) as well as phase II (glutathione-S-transferase and UDP-glucuronosyl transferase) biotransformation enzymes; and reduced glutathione (GSH) (in forestomach and hepatic tissues) during/after intra-gastric instillations of B(a)P in murine forestomach tumorigenesis bioassay protocol. The activities of phase I biotransformation enzymes were found to increase, whereas a decrease in GSH content as well as glutathione-S-transferase was observed in mice receiving only B(a)P during as well as 2 weeks after B(a)P instillations. The activity of UDP-glucuronosyltransferase decreased during B(a)P instillations, whereas after the latter, the activity increased when compared with the control mice. However, in mice that received AAILE along with B(a)P instillations, a decrease in phase I enzymes was accompanied by an increase in phase II enzymes as well as GSH contents. Only AAILE treatment reduced the activities of phase I biotransformation enzymes and enhanced the GSH contents as well as the activities of phase II enzymes. Observations of the present study seem to be quite significant and (when taken together with our earlier findings) provides evidence for A. indica mediated modulation of the peri-initiation phase of the process of forestomach tumorigenesis.     

Antipyretic effect of Azadirachta indica in bacteria endotoxin induced fever in the rat

Fever was induced in rats with a single i.p. injection of 30 μg/kg E. coli endotoxin. The effect of the alcohol leaf extract of Azadirachta indica was investigated in this model. Pretreatment of rats with the leaf extract (125–375 mg/kg) did not significantly reduce endotoxin-induced fever. The rectal temperature remained significantly high. In contrast, administration of the same doses of the extract during the early phase of fever development (during temperature rise) produced a significant fall in the rectal temperature to near normal. The maximum rise was 1.0°C which later dropped to 0.18°C, and was sustained even beyond the experimental session. The data obtained suggest the beneficial antipyretic effect of the leaf extract of Azadirachta indica in bacteria endotoxin induced fever. © 1998 John Wiley & Sons, Ltd.     

蛇莓乙酸乙酯萃取物的化学成分

目的:研究蛇莓乙酸乙酯萃取物的化学成分.方法:以95%乙醇回流提取,对提取液的乙酸乙酯萃取物采用正相硅胶、反相硅胶、Sephadex LH-20柱色谱和高效液相色谱进行分离纯化,并根据理化性质和波谱数据鉴定化合物的化学结构.结果:分离得到13个化合物,分别鉴定为蔷薇酸(1),arjunic acid(2),对羟基桂皮酸(3).芹菜素(4),山柰酚(5),2α-羟基乌苏酸(6),2α-羟基齐墩果酸(7),委陵菜酸(8),刺梨苷(9),翻白叶苷A(10),野蔷薇苷(11),紫云英苷(12)和异槲皮苷(13).结论:化合物2～5、10、12均为首次从蛇莓属中分离得到.     

复方鬼辣椒根液的长期毒性实验

目的观察复方鬼辣椒根口服液(以下简称复辣液)对小鼠长时间给药可能产生的毒性反应,为临床应用提供毒理学依据。方法取复辣液分别相当于人日用量的60倍、30倍、15倍,经90天小鼠灌胃给药,观察给药过程中、给药末期及停药恢复期动物的一般状况,主要血液学、血液生化学和脏器病理组织变化。结果各实验期动物一般状况良好,体重、脏器主要血液学指标、血液生化学指标与空白对照组基本一致,对性别、体重无影响,解剖主要脏器未见明显内眼可见的病理性改变。结论该品长期服用无明显毒性,停药后未见药物延迟性毒性反应。     

大孔树脂纯化马兰总三萜工艺研究

目的 研究大孔树脂纯化马兰总三萜的工艺。方法 以齐墩果酸为对照品,以吸附量和解吸率作为效果评定指标,筛选出对马兰中总三萜纯化效果最好的大孔树脂,并对上样量、上样液浓度、洗脱速率、洗脱乙醇浓度及用量参数进行考察,筛选出最佳工艺条件。结果 HPD-500的吸附量为0.495 5 mg/mL,解吸率为30.47%,效果最佳;最佳上样液浓度：0.02 g/mL;流速：2 mL/min;洗脱剂：70%乙醇;洗脱体积：2倍柱体积。纯化后马兰三萜含量可达到17.81%。结论 HPD-500大孔树脂在所得工艺条件下可以对马兰中总三萜进行较好的纯化。     

Lack of in vivo embryotoxic and genotoxic activities of orally administered stem bark aqueous extract of Mangifera indica L. (Vimang)

Mango (Mangifera indica L.) stem bark aqueous extract (MSBE) is a new natural product with antioxidant, anti-inflammatory and immunomodulatory effects known by the brand name of its formulations as Vimang^®. Previously, the oral toxicity studies of the extract showed a low toxicity potential up to 2000 mg/kg. This work reports the results about teratogenic and genotoxicologic studies of MSBE. For embryotoxicity study, MSBE (20, 200, or 2000 mg/kg/day) was given to Sprague–Dawley rats by gavage on days 6–15 of gestation. For genotoxicity, MSBE was administered three times during 48 h to NMRI mice. Cyclophosphamide (50 mg/kg) was used as a positive control. No maternal or developmental toxicities were observed when the rats were killed on day 20th. The maternal body-weight gain was not affected. No dose-related effects were observed in implantations, fetal viability or external fetal development. Skeletal and visceral development was similar among fetuses from all groups. No genotoxicity was observed in bone marrow erythrocytes and liver cells after administration. MSBE appears to be neither embryotoxic nor genotoxic as measured by bone marrow cytogenetics in rodents.