Efficiency of Coleus aromaticus extract in modifying cyclophosphamide and mitomycin-C induced clastogenicity in mouse bone marrow cells

The anticlastogenic potency of the ethanolic extract of a medicinal plant, C. aromaticus was investigated by taking bone marrow chromosomal aberration assay and micronucleus (MN) test as the test parameters. Swiss albino mice were fed orally with different doses (10,15, 25, 50 and 100 mg/kg body weight) of ethanolic extract for 7 days and on the 7th day, two doses each of anticancer drugs cyclophosphamide (CP; 25 and 50 mg/kg body weight) and mitomycin-C (MMC; 4 and 8 mg/kg body weight) were injected, ip, to different groups of animals. Bone marrow MN preparations were made at 24 and 48 hr time intervals. Coleus extract reduced CP and MMC induced MN and lower doses of the extract were found to be more effective than higher doses. The effective doses of extract in MN test were selected to study the anticlastogenic effects against CP (25 and 50 mg/kg body weight) and MMC (2 and 4 mg/kg body weight) induced chromosomal aberrations. The results indicate the protective effect of C. aromaticus against CP and MMC induced cytogenetic damage.     

Modifying effect of iron on lead-induced clastogenicity in mouse bone marrow cells

Essential trace elements such as iron (Fe) are known to interact with nonessential metals like lead (Pb), influencing its metabolism. Ferric chloride and lead nitrate were administered intraperitoneally to Swiss albino miceMus musculus singly and successively, with or without a time gap of 1 h, to study the degree of protection, if any, afforded by iron against the clastogenic effects induced by Pb in bone marrow cells. A decrease in the frequency of lead-induced chromosomal aberrations was observed when Fe was given together with or prior to Pb administration.     

Protection against radiation clastogenecity in mouse bone marrow by Phyllanthus niruri

The effects of aqueous (PnAq) and alcoholic (PnA1 extract (50-250 mg/kg) of P. niruri on in vivo gamma radiation induced chromosome aberration and in vitro antioxidant activity (50-500 microg/ml) were studied. The antioxidant activity was studied by measuring inhibition of hydroxyl radicals generated by the fenton reaction along with pro-oxidant and iron chelating ability. PnA1 showed highly significant in vitro free radical scavenging ability when compared to DMSO above 250 microg/ml concentration. PnAq showed significant pro-oxidant activity while PnA1 was devoid of it at the tested concentrations. Exposure to gamma radiation (4 Gy) caused 29.10 % increase in the frequency of chromosomal aberrations. Administration of PnA1 (250 mg/kg) showed highly significant decrease in chromosomal aberrations compared to radiation treated group. Radioprotective potential of alcoholic extract was found to be more effective than the aqueous extract. Qualitative phytochemical investigation of PnAq and PnA1 revealed the presence of sugars, flavonoids, alkaloid, lignans, polyphenols, tannins, coumarins and saponins. Higher radioprotective effect of the alcoholic extract may be attributed to rich presence of antioxidant polyphenolic compounds.     

Lithium attenuates lead induced toxicity on mouse non-adherent bone marrow cells

Lead is a poisonous heavy metal that occurs in all parts of environment and causes serious health problems in humans. The aim of the present study was to investigate the possible protective effect of lithium against lead nitrate induced toxicity in non-adherent bone marrow stem cells. Trypan blue and MTT assays represented that exposure of the cells to different concentrations of lead nitrate decreased viability in a dose dependent manner, whereas, pretreatment of the cells with lithium protected the cells against lead toxicity. Lead reduced the number and differentiation status of bone marrow-derived precursors when cultured in the presence of colony stimulating factor (CSF), while the effect was attenuated by lithium. The cells treated with lead nitrate exhibited cell shrinkage, DNA fragmentation, anion superoxide production, but lithium prevented lead action. Moreover, apoptotic indexes such as PARP cleavage and release of HMGB1 induced by lead, were protected by lithium, suggesting anti-apoptotic effect of lithium. Immunoblot analysis of histone H3K9 acetylation indicated that lithium overcame lead effect on acetylation. In conclusion, lithium efficiently reduces lead toxicity suggesting new insight into lithium action which may contribute to increased cell survival. It also provides a potentially new therapeutic strategy for lithium and a cost-effective approach to minimize destructive effects of lead on bone marrow stem cells.     

Phyllanthus emblica fruit extract attenuates lipid metabolism in 3T3-L1 adipocytes via activating apoptosis mediated cell death

BackgroundPhyllanthus emblica L. (Indian gooseberry) is widely used in the Ayurveda for thousands of years to treat health complications including disorders of the immune system, diabetes, and obesity.PurposeFor the first time, our study aims to demonstrate the molecular mechanisms of the fruit extract of Phyllanthus emblica (PEFE) involved in the promotion of fat cell apoptosis and alleviation of adipogenesis.MethodsThe active constituents from PEFE were identified using high performance liquid chromatography-mass spectrometry (HPLC-MS). We carried out the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay to evaluate the cytotoxic effects of PEFE using 3T3-L1 pre-adipocytes. The colonogenic assay was carried out to determine the inhibitory effect of 3T3-L1 adipocytes after PEFE treatment. In addition, inhibition of pancreatic lipase activity was performed and the lipolytic activity of PEFE and digallic acid was compared with the well-known standard drug orlistat. Besides, the molecular interaction and ligand optimization between digallic and adipogenesis/apoptosis markers were also carried out. Furthermore, to confirm fat cell apoptosis we have used several detection methods that includes Hoechst staining, PI staining, Oil staining and qPCR respectively.ResultsDigallic acid was identified as a major component in the PEFE. The IC₅₀ values of digallic acid and PEFE were found to be 3.82 µg/ml and 21.85 µg/ml respectively. PEFE and digallic acid showed significant anti-lipolytic activity compared to the standard drug orlistat. In the mature adipocytes, PEFE significantly decreased triglyceride accumulation by downregulating adiponectin, PPARγ, cEBPα, and FABP4 respectively. We further analyzed the expression of apoptosis related genes upon PEFE treatment. Apoptotic process initiated through upregulation of BAX and downregulation of BCL2 resulting in an increased caspase-3 activity. In addition, we have also confirmed the apoptosis and DNA fragmentation in 3T3-L1 cells using Hoechst, PI and TUNEL assays.ConclusionPEFE negatively regulates adipogenesis by initiating fat cell apoptosis and therefore it can be considered as a potential herbal medicinal product for treating obesity.     

Histamine production by alloantigen-activated mouse bone marrow cells

Histamine's contribution to the manifestations associated with graft-versus-host disease (GVHD) and/or hybrid resistance is unknown. Thus, we initiated studies to see whether or not mouse bone marrow cells could produce histamine upon alloantigen stimulation. Irradiated allogeneic spleen cells were shown to stimulate bone marrow cells to produce and secrete high levels of histamine. During 7 days of culture there was only a marginal increase in cell-associated histamine while the amount of histamine in the supernatant increased 10- to 20-fold. Optimal histamine production was dependent upon Lyt 1+2+ T cells resident in the bone marrow. Further, bone marrow cells from Nude mice failed to produce high levels of histamine following alloantigen stimulation. Soluble factors produced by alloantigen-stimulated bone marrow cells or by Con A-stimulated rat spleen cells induced high levels of histamine production in bone marrow cells in the absence of alloantigen. We suggest that histamine production by alloantigen-activated bone marrow cells may modulate immune functions following bone marrow transplantation.     

Radioprotective effects of Dragon's blood and its extract against gamma irradiation in mouse bone marrow cells

PurposeThe radioprotective effects of Dragon's blood (DB) and its extracts (DBE) were investigated using the chromosomal aberrant test, micronucleus and oxidative stress assay for anti-clastogenic and anti-oxidative activity.Materials and methodsAdult BALB/C mice were exposed to the whole body irradiation with 4 Gy ⁶⁰Co γ-rays. DB and DBE were administered orally once a day from 5 days prior to irradiation treatment to 1 day after irradiation. The mice were sacrificed on 24 h after irradiation. The cells of bone marrow were measured by counting different types of chromosomal aberrations and the frequency of micronuclei. Oxidative stress response was carried out by analysis of serum from blood.ResultsDB and DBE significantly decreased the number of bone marrow cells with chromosome aberrations after irradiation with respect to irradiated alone group. The administration of DB and DBE also significantly reduced the frequencies of micronucleated polychromatic erythrocytes (MPCE) and micronucleated normochromatic erythrocytes (MNCE). In addition, DB and DBE markedly increased the activity of antioxidant enzymes and the level of antioxidant molecular. Malondialdehyde (MDA) and nitric oxide (NO) levels in serum were significantly reduced by DB and DBE treatment.ConclusionsOur data suggested that DB and DBE have potential radioprotective properties in mouse bone marrow after ⁶⁰Co γ-ray exposure, which support their candidature as a potential radioprotective agent.     

Immunoglobulin-positive cells in mouse bone marrow]

The content of Ig-bearing lymphocytes and their precursors in the mouse bone marrow was investigated 6 and 36 hours after the hydroxyurea treatment. Some increase of the B-cell content takes place in the trated bone marrow. Dividing and non-dividing B-cell precursors, except the stem cells, were practically absent.     

Identifying superoxide-sensitive progenitor cells in mouse bone marrow

Macrophage progenitor cells in bone marrow, that develop into attached colonies in liquid culture medium, contain a fraction of cells sensitive to photochemically generated superoxide radicals. This fraction varies from one animal to another. Populations of cells containing the superoxide-sensitive fraction show a greater sensitivity to X-rays than do populations in which this fraction has been photochemically inactivated. The change in radiosensitivity was proportional to the superoxide-sensitive fraction.     

余甘子采后软腐病拮抗菌的筛选、鉴定及其防效

【背景】由青霉菌引起的软腐病是余甘子采后贮藏期的主要病害,造成了经济损失严重。【目的】筛选对余甘子软腐病菌具有良好拮抗效果的放线菌菌株,并初步评价拮抗菌无菌发酵液的防治效果。【方法】以PenicilliumchoerospondiatisDQ23为指示菌,采用琼脂块法结合生长速率法进行拮抗菌株的筛选,并采用形态学特征、生理生化特性结合16S rRNA基因序列进行分类鉴定,并采用浸果法初步测定其无菌发酵液对余甘子软腐病发生及果实贮藏品质的影响。【结果】菌株SC-15对青霉P.choerospondiatis DQ23有较强的拮抗作用,拮抗圈直径为18.70 mm,20%无菌发酵液对软腐病菌的抑制率可达87.8%。根据菌株形态学和生理生化特性,结合16SrRNA基因序列分析将菌株SC-15鉴定为维及尼链霉菌(Streptomycesvirginae)。菌株SC-15无菌发酵液能降低余甘子的腐烂率和腐烂指数,延缓可溶性固形物和可滴定酸的降低,其中75%发酵液浸果效果最好,贮藏8d后,果实腐烂率为53.3%,果实腐烂指数为35.2%,可溶性固形物为4.8%,可滴定酸为6.25mg/g。【结论】...     

Stimulation by normal and leukemic mouse sera of colony formation in vitro by mouse bone marrow cells

Using a modification of the agar gel method for bone marrow culture, serum from various strains of mice has been tested for colony stimulating activity. Ninety percent of sera from AKR mice with spontaneous or transplanted lymphoid leukemia and 40–50% of sera from normal or preleukemic AKR mice stimulated colony formation by C57B1 bone marrow cells. Sera from 6% of C3H and 30% of C57B1 mice stimulated similar colony formation. The incidence of sera with colony stimulating activity rose with increasing age. All colonies were initially mainly granulocytic in nature but later became pure populations of mononuclear cells. Bone marrow cells exhibited considerable variation in their responsiveness to stimulation by mouse serum. Increasing the serum dose increased the number and size of bone marrow cell colonies and with optimal serum doses, 1 in 1000 bone marrow cells formed a cell colony. Preincubation of cells with active serum did not stimulate colony formation by washed bone marrow cells. The active factor in serum was filterable, non-dialysable and heat and ether labile.     

Identification of parathyroid hormone-regulated proteins in mouse bone marrow cells by proteomics

The ability of parathyroid hormone (PTH) to enhance bone formation has recently been exploited in the treatment of osteoporosis. Several studies have suggested that the activation of bone marrow stromal cells could be preceded to show the anabolic effect of PTH on bone formation, but little is known of PTH-regulated proteins in bone marrow cells. Therefore, protein profiling in the intermittent PTH-treated bone marrow cells was evaluated using proteomics. Daily treatment for 5 days consisting of subcutaneous injection of either 150 microg/kg per day of mouse PTH (1-84) or vehicle (0.9% normal saline) was performed on the ICR mouse. At the end of the treatment period, bone marrow cells were separated and used in proteomics. The expression levels of seven proteins including vimentin were decreased, but those of four proteins including calreticulin and thioredoxin domain containing 7 protein (Txnde7) were increased. Among these, the decrease of vimentin and the increase of both calreticulin Txnde7 in mRNA levels were confirmed by semi-quantitative RT-PCR. In PTH-treated mouse MC3T3-E1 osteoblast cells, mRNA expression levels were not totally consistent with the results observed in proteomics. In conclusion, the differentially expressed proteins in bone marrow cells depending on PTH could be highly linked to the differentiation of osteoprogenitor cells in the bone marrow into preosteoblast cells.     

Formation of eosinophilic-like granulocytic colonies by mouse bone marrow cells in vitro

Formation of granulocytic and macrophage colonies in agar cultures of mouse marrow or spleen cells was stimulated by the addition of medium from pokeweed mitogen-stimulated cultures of mouse spleen cells (PKW-CM). Approximately 5% of the colonies developing were large, dispersed granulocytic colonies (DG-colonies) composed of cells with eosinophilic cytoplasmic granules. The capacity to stimulate DG-colonies was shown by media conditioned by PKW-treated lymphoid and peritoneal cells but not by other cells or organ fragments. Velocity sedimentation studies indicated that cells generating DG-colonies were separable from cells generating regular granulocytic or macrophage colonies. DG-colonies did not survive if transfered to cultures containing other forms of CSF. The active colony stimulating factor in pokeweed mitogen-conditioned medium which stimulates DG-colony formation was antigenically distinct from the factor stimulating granulocytic and macrophage colony formation, was separable electrophoretically from the latter factor and on gel filtration had an apparent molecular weight of 50,000. Although the cells in DG-colonies have not been established to be eosinophils, DG-colonies represent an interesting new system for analysing further aspects of the control of growth and differentiation in hemopoietic populations.     

余甘子总黄酮超声波辅助提取工艺的优化

以余甘子干粉为材料,利用超声波辅助提取余甘子总黄酮。单因素试验分析液料比、乙醇浓度、超声时间、超声功率对余甘子总黄酮提取率的影响,在此基础上,利用响应面分析法对提取工艺进行优化。结果表明,余甘子总黄酮最佳提取条件为：液料比82:1(V/W)、乙醇浓度45%、超声时间41 min、超声功率210 W,在此条件下余甘子总黄酮的实际提取率为6.12%。     

Clastogenic effects of acrylamide in mouse bone marrow cells

Acrylamide, known to induce dominant-lethal mutations (Shelby et al., 1986; Smith et al., 1986) and heritable translocations (Shelby et al., 1987) in rodent germ cells, was hitherto a questionable clastogen in rodent bone marrow (Shiraishi, 1978). Therefore, it was tested for chromosomal aberrations in mouse bone marrow cells, spermatogonia and by the micronucleus test. The intraperitoneally injected doses ranged from 50 to 150 mg/kg. In the chromosomal bone marrow test and the micronucleus assay positive results were obtained with acrylamide, and in the latter test the effect increased linearly with dose. Chromosomal aberrations were not induced in differentiating spermatogonia by the acute acrylamide treatment. Cisplatin was used as a positive control and gave the expected positive response in all 3 tests. The present results demonstrate that acrylamide is no exception among clastogens. It breaks chromosomes not only in mammalian germ cells but also in somatic cells.     

余甘子采后软腐病菌的分离鉴定及其细胞壁水解酶活性

【背景】药食同源的余甘子果实在采后贮藏过程中极易软腐变质,严重影响其品质和经济价值。【目的】明确引起余甘子果实软腐病的病原菌种类及其生长特性和产细胞壁水解酶活性,为余甘子采后软腐病的控制及延长其贮藏期奠定基础。【方法】采用组织块分离法从采后发病的余甘子果实分离病原菌,按照科赫法则确定分离菌株的致病性;采用形态学特征结合rDNA-ITS序列分析对病原菌进行鉴定,测定病原菌菌丝生长和产孢特性,检测产胞外细胞壁水解酶活性。【结果】从具有软腐症状的余甘子果实中分离得到32株真菌,其中菌株DQ23是余甘子采后软腐病的致病菌,通过形态特征结合rDNA-ITS序列将其鉴定为Penicillium choerospondiatis。其菌丝在酵母膏葡萄糖琼脂培养基(YDA)上生长最快,在马铃薯蔗糖琼脂培养基(PSA)上产孢最多。该菌能有效利用多种碳、氮源,适宜产孢的碳源为蔗糖、葡萄糖,氮源为蛋白胨、牛肉膏、酵母膏。菌丝生长的最适温度和pH范围分别为25°C和3.0-5.0,产孢的最适温度和pH范围分别为25°C和4.0-7.0。光照均利于菌丝生长和产孢。该菌具有分解果胶、纤维素的能力,无分解蛋白质、鞣质的能力。【结论】Penicillium choerospondiatis是余甘子果实软腐病的病原菌,研究结果为该病害的防控奠定了基础。     

Study of pharmacodynamic interaction of Phyllanthus emblica extract with clopidogrel and ecosprin in patients with type II diabetes mellitus

BackgroundDiabetes mellitus is associated with oxidative stress which impairs the platelet function. Phyllanthus emblica extract a rich source of vitamin C plays an important role in scavenging free radicals. The effect of vitamin C on platelet aggregation in healthy and coronary artery disease patients has been demonstrated. The present study attempts to study the pharmacodynamic interactions of P. emblica extract with clopidogrel and ecosprin.Materials and methodsThis was a randomized open label crossover study of 10 type II diabetic patients. The dosage schedules were either single dose of 500 mg P. emblica extract or 75 mg clopidogrel or 75 mg ecosprin or 500 mg P. emblica + 75 mg clopidogrel or 500 mg P. emblica + 75 mg ecosprin. After single dose study and washout period, patients received either 500 mg P. emblica extract twice daily or 75 mg clopidogrel or 75 mg ecosprin once daily or combinations for 10 days. Platelet aggregation was measured at baseline and at 4 h of treatment after single and multiple dose study along with recording of bleeding and clotting time.ResultsAfter single and multiple dose administration of the three treatments and with combinations there was statistically significant decrease of platelet aggregation compared to baseline. Further, the mean percent inhibition of platelet aggregation was significant, when compared between single and multiple doses of P. emblica. The bleeding and clotting time was prolonged with single and multiple dose administration of all treatments compared to baseline. All treatments were well tolerated.ConclusionP. emblica extract demonstrated significant antiplatelet activity with both single and multiple dose administration.