Inhibitory Effect of Flavonoid Glycosides from Chlorophytum comosum on Nasopharyngeal Carcinoma 5-8F Cells and Its Mechanism

[Objectives]To study the inhibitory activity of two flavonoid glycosides isolated from Chlorophytum comosum Laxum R.Br on human nasopharyngeal carcinoma(NPC)cell line 5-8F in vitro and its mechanism.[Methods]The flavonoid glycosides were isolated and purified from the ethanol alcoholic extract of the roots of Liliaceae plant Chlorophytum comosum by silica gel column chromatography,macroporous resin column chromatography,Sephadex LH-20,and reverse column chromatography(ODS).The inhibitory activity of flavonoid glycosides on human nasopharyngeal carcinoma cells was analyzed by CCK-8 method,and the potential mechanism was preliminarily analyzed by molecular docking.[Results]Two flavonoid glycosides were identified as isovitexin 2″-0-rhamnoside and 7-2″-di-O-β-glucopyranosylisovitexin.Two flavonoid glycosides showed promising inhibitory effect on human nasopharyngeal carcinoma cell line 5-8F,with IC_(50) values of 24.8 and 27.5μmol/L,respectively.Molecular docking results showed that the potential targets of two flavonoid glycosides include CyclinD1,Bcl-2β-Catenin,ILK,TGF-β,in addition,two glycosides showed higher predicted binding affinity towards CyclinD1,which verifies the cytotoxicity of the two compounds on human nasopharyngeal carcinoma cell line 5-8F in vitro.[Conclusions]Two flavonoid glycosides are the active molecules in Chlorophytum comosum that can inhibit the proliferation of human nasopharyngeal carcinoma cells,and have the potential to be used in the research and development of anti nasopharyngeal carcinoma drugs.

吊兰属植物Chlorophytum tuberosum的化学成分及其对血小板聚集的作用

[目的]研究Chlorophytum tuberosum50%乙醇提取物中的化学成分及其对血小板聚集的作用。[方法]运用各种柱色谱及制备型高效液相等对化学成分进行分离纯化,综合运用NMR、LC-MS等波谱技术鉴定化合物结构;以ADP为激动剂,阿司匹林为阳性对照,初步测定了不同极性洗脱物及部分化合物对血小板聚集的影响。[结果]共分离得到11个化学成分,分别为:新替告皂苷元(1)、(25S)-曼诺皂苷元(2)、新海柯皂苷元(3)、新海柯皂苷元-3-O-β-D-葡萄糖-(1→2)-[β-D-葡萄糖-(1→3)]-β-D-葡萄糖-(1→4)-β-D-半乳糖苷(4)、新吉托皂苷元-3-O-β-D-葡萄糖-(1→2)-[β-D-葡萄糖-(1→3)]-β-D-葡萄糖-(1→4)-β-D-半乳糖苷(5)、新吉托皂苷元-3-O-β-D-葡萄糖-(1→2)-[β-D-葡萄糖-(1→4)]-β-D-葡萄糖-(1→4)-β-D-半乳糖苷(6)、新吉托皂苷元-3-O-β-D-葡萄糖-(1→2)-[β-D-木糖-(1→3)]-β-D-葡萄糖-(1→4)-β-D-半乳糖苷(7)、大叶吊兰苷C(8)、大叶吊兰苷E(9)、β-谷甾醇(10)和胡萝卜苷(11)。[结论]化合物1为吊兰属首次分离,化合物2为百合科首次分离,其他化合物均为首次从该植物中分离得到;70%乙醇洗脱物及化合物6、7有诱导血小板聚集的活性,而95%乙醇洗脱物有抑制血小板聚集的活性。

Chlorophytoside A, a New Labdane Diterpene Glycoside from Chlorophytum laxum

A new labdane-type diterpene glycoside 1 , chlorophytoside A, had been isolated from Chlorophytum laxum R.Br. The structure had been elucidated as (10S)-6α-hydroxy- labda-8,13-dien-15,16-olide 3R-O-β-D-glucopyranoside on the basis of chemical and spectroscopic data.

Safety assessment of Chlorophytum alismifolium tuber extract (Liliaceae):Acute and sub-acute toxicity studies in Wistar rats

To explore the toxicological profile of methanol extract of Chlorophytum alismifolium (MECA) tubers in Wistar rats. Methods: MECA was subjected to acute and sub-acute studies which were conducted according to Organization for Economic Co-operation and Development (OECD 425 and 407 guidelines respectively). In the acute toxicity experiment, a limit test (5000 mg/kg) was administered to five rats and monitored for 2 weeks. The sub-acute studies were conducted on 4 groups of rats. The first group served as control, while the 2nd, 3rd and 4th groups received MECA (150, 300 and 600 mg/kg respectively). The treatments were given orally and daily for 4 weeks. At the end of the experiment (29th day), the animals were euthanized to obtain blood samples and organs for haematological, biochemical and histological evaluations. Results: Acute toxicity study showed that the oral median lethal dose was >5000 mg/kg. In the sub-acute studies, the results showed no significant (P>0.05) changes in the haematological, hepatic and renal indices compared to control animals. In the fourth week, a significant (P<0.01) increase in body weight of the rats was observed at 150 mg/kg and 600 mg/kg compared to week one. However, there were no major changes in the organ/body weights of the rats. Histological examination of the kidney showed slight glomerular adhesion and tubular distortion. Moderate hepatic necrosis was observed at 150 mg/kg and 300 mg/kg. Conclusions: The results of this research revealed that the MECA tubers is virtually non-toxic after acute administration and it has low sub-acute toxicity potential in rats.

Effect of Intravital Staining on Leaf Surface Coloring and Plant Carbon and Nitrogen Nutrition in Chlorophytum comosum var. variegatum

Using potted seedlings of Chlorophytum comosum var. variegatum as the experimental materials, the effect of 2.0 mmol/L methyl orange （ Treatment T1 ）, 1.0 mmol/L methyl violet （ Treatment T2 ） and 1.0 mmol/L neutral red （ Treatment T3 ） on the biomass, root-shoot ratio, leaf color indices, plant carbon and nitrogen nutrition were studied. The results showed that the biomass of Treatment T3 was significantly greater than that of treatments T1 and CK. The root-shoot ratio decreased significantly in treatments T1, T2 and T3 , and the decrease in T3 was most obvious. In all the three treatments with coloring agent, a ^＊ , b ^＊ and L ^＊ values were increased gradually, C value were decreased, H0 and CIRG were increased, and the leaves were pink. In addition, the contents of chlorophyll a, chlorophyll b, chlorophyll a ＋ b and carotenoid were significantly decreased. The contents of soluble sugar and starch were also decreased, and the decrease in Treatment T2 was most significant. The contents of soluble protein and total nitrogen were increased, and the increase was most dramatic in Treatment T3. The carbon to nitrogen ratio was decreased. The results proved that staining can improve the ornamental value of indoor plants, despite its effects on plant carbon and nitrogen nutrition of C. comosum vat. variegatum, dyeing.

ISSR Analysis of Chlorophytum Treated by Three Kinds of Chemical Mutagen

Chlorophytum leaves were treated with three kinds of chemical mutagen, EMS, DES and NaN3 with different concentration to obtain the variation materials with excellent properties. The results showed that the genetic similarity coefficient of variation plants with EMS treatment was between 0.648 and 0.868, with NaN3 treatment was between 0.598 and 0.859, and with DES treatment was between 0.668 and 0.904, of which the mutagenic effects with 0.8% EMS, 250 mg. L-1 NaN3 and 0.3% DES on chloro- phytum were the best. ISSR molecular marker technique was used to analyze their genetic diversities. Total 392 polymorphic bands were obtained through 18 ISSR primers. Polymorphic ratio was 72.4%, which showed that DNA mutation took place in various degrees

Regeneration and Genetic Fidelity Analysis of <i>Chlorophytum borivilianum</i>Using Flower Stalk as Explant Source

Chlorophytum borivilianum is a critically endangered plant well known for its medicinal properties for diabetes mellitus, diarrhea, arthritis, sterility, and erectile dysfunction, etc. Due to low viability and long dormancy of seeds, in vitro regeneration is required for large scale cultivation of this plant. In the present study, direct plant regeneration was optimized using flower stalk as explant. Nodal segments of flower stalk were sterilized and kept for direct regeneration on different combinations of BAP and KIN supplemented media. The highest, 15.27 ± 1.14 number of shoots were produced on medium containing BAP (2 mg/L) per nodal segment. The multiple shoot clumps regenerated from flower stalk were separated carefully and kept on rooting media. A maximum of 16.87 ± 1.53 roots per plant was observed in MS media having 0.5 mg/L of NAA. The rooted plantlets were shifted into the pot containing soilrite for hardening and acclimatization. The genetic stability of hardened plants was confirmed by start codon targeted, and inter simple sequence repeats molecular markers. All the 18 randomly selected plantlets showed similar genetic homogeneity to the mother plant. It is the first report on in vitro regeneration along with the genetic fidelity analysis of the regenerated plantlets from flower Stalk of C. borivilianum. As the standardized method of regeneration and mass multiplication is quite efficient and genetically stable, the protocol will be useful for the large-scale production of C. borivilianum to meet the market demand.

Effect of Chlorophytum Borivilianum Santapau and Fernandes on Sexual Dysfunction in Hyperglycemic Male Rats

Objective:To investigate the effects of Chlorophytum borivilianum(CB) on sexual dysfunction, loss of body weight,and lack of libido in hyperglycemic rats induced with streptozotocin or alloxan.Methods: Wistar strain male albino rats were divided into five groups of six animals each:the control group(2% polyvinylpyrollidone solution),the streptozotocin control group(50 mg/kg),the alloxan control group(100 mg/kg), the streptozotocin + CB treated group(200 mg/kg),and the alloxan + CB treated group(200 mg/kg).O...

Negative Phototropism of Chlorophytum comosum Roots and Their Mechanisms

The aerial roots of Chlorophytum comosum were grown hydroponically,allowing us to study the performance and mechanism of negative phototropism. The results of this study were as follows. All the adventitious roots and their branch roots bent away from light with a maximum curvature of approximately 88.5°. Blue-violet light prominently induced negative phototropism while red light had no effect. The root cap was the site of photo perception. Roots with shaded or divested root caps exposed to unilateral light showed no negative phototropism,but resumed their original characteristics when the shade was removed or when new root caps grew. The curvature increased when the light intensity ranged 0–110 μmol·m^(-2)·s^(-1). The negative phototropism curvature could be promoted by exogenous CaCl_2 but was inhibited by exogenous LaCl_3; exogenous CaCl_2 could reduce the inhibitory effect of LaCl_3. Unilateral light induced the horizontal transport of IAA from the irradiated side to the shaded side,resulting in an unequal distribution of IAA in both the sides,leading to negative phototropism. The horizontal transport of IAA was promoted by exogenous Ca^(2+) but inhibited by exogenous La^(3+).

Saponins isolated from roots of Chlorophytum borivilianum reduce acute and chronic inflammation and histone deacetylase

OBJECTIVE： The roots of Chlorophytum borivilanum are used in traditional medicine for the treatment of arthritis and inflammation. The aim of the work was to evaluate the anti-inflammatory activity of isolated saponins from Chlorophytum borivilianum （ISCB）. METHODS： The ISCB was screened using the carrageenan-induced paw edema, histamine-induced paw edema, cotton pellet-induced granuloma, and Freund＇s adjuvant-induced arthritis in rats at orally administered doses of 3, 10, and 30 mg/kg. Effect of ISCB on histone deacetylase （HDAC） level was measured by the HDAC assay at the highest dose （30 mg/kg）. RESULTS： The results showed that the ISCB significantly reduced carrageenan-induced inflammation, histamine-induced inflammation, cotton pellet-induced granuloma and Freund＇s adjuvant-induced arthritis in rats. The ISCB at a dose of 30 mg/kg significantly inhibited HDAC level in rat paw tissue. CONCLUSION： It is concluded that saponins isolated from roots of C. borivilianum possess anti- inflammatory and anti-arthritic properties. ISCB may act by inhibiting histamine, prostaglandin and HDAC. This suggests that ISCBs have potential for therapeutic use in the treatment of inflammation and arthritis

Antihyperglycaemic activity of ethylacetate extract of Chlorophytum alismifolium in type 2 diabetes:The involvement of peroxisome proliferator-activated receptor-γand dipeptidyl peptidase-4

Objective:This research is to investigate the antihyperglycaemic activity and the underlying mechanisms of action of the ethylacetate extract of Chlorophytum alismifolium(EACA)tubers in a type 2 diabetes model.Methods:The tubers were processed and sequentially extracted in hexane followed by ethylacetate,using a Soxhlet apparatus,and subjected to gas chromatography-mass spectrometry(GC–MS).The acute toxicity of EACA was investigated in albino Wistar rats.An antihyperglycaemic study was carried out using high-fat diet(pelletized diet and margarine in the ratio of 10:1 and 20%fructose solution)and streptozotocin-induced hyperglycaemic Wistar rats.The effects of the extract(150,300 and 600 mg/kg)on blood glucose level,insulin,peroxisome proliferator-activated receptor-c(PPAR-c)and dipeptidyl peptidase-4(DPP-4)were evaluated using enzyme-linked immunosorbent assay.Results:The oral median lethal dose in Wistar rats was estimated to be>5000 mg/kg.Treatment with EACA at all doses significantly reduced the fasting blood glucose levels,compared to the hyperglycaemic control,and over time.Administration of EACA increased the serum insulin and PPAR-c levels while decreasing DPP-4 levels.GC–MS analysis revealed the presence of 13 compounds,with isothiazole and isoxazolidine covering total area of 24.6%and 22.69%,respectively.Conclusion:The findings from this study showed that EACA has important compounds with beneficial effect in type 2 diabetes and acts by increasing insulin secretion and PPAR-c level and decreasing DPP-4 activity.

Antioxidant and modulatory role of Chlorophytum borivilianum against arsenic induced testicular impairment

Arsenic has a suppressive influence on spermatogenesis and induces impairment in male reproductive system due to oxidative stress. The present study was aimed to test the arsenic induced toxicity and protection by Chlorophytum borivilianum. The effect of sodium arsenite （4 mg/（kg body weight （bw）.day）） via double distilled water without or with C. borivilianum （800 mg/（kg bw.day）） was evaluated in Swiss albino mice for 30 days. The radical scavenging activity of the aqueous C. borivilianum root extract was measured using DPPH （1,1-diphenyl-2-picryl hydrayzyl） radical. Qualitative assessment of various cell types in the testis, sperm count and motility, testicular activity of lipid peroxidation （LPO）, reduced glutathione （GSH）, acid and alkaline phosphatase, cholesterol and serum testosterone were monitored. Arsenic treatment showed a significant increase in LPO, acid and alkaline phosphatase, cholesterol and decrease in sperm count, sperm motility, GSH and serum testosterone. Combined treatment showed significant decrease in LPO, acid and alkaline phosphatase, cholesterol and elevation in sperm count, sperm motility, GSH and serum testosterone. Testicular histopathology showed that C. borivilianum had reduced degeneration of germ cell in the seminiferous tubules and loss of sperms induced by arsenic intoxication. The results thus led us to conclude that administration of C. borivilianum root extract is found to be protective against arsenic induced toxicity.

Structure and Activity of a New Sapogenin from Chlorophytum laxum R. Br.

A new steroidal sapogotin named 25-R-spirosta-3,5-dien-12β-ol（1） was isolated from the dried roots of Chlorophytum laxum R. Br. along with five known compounds, namely, diosgenin（2）, stigmasterol（3）, β-sitosterols（4）, estigmasterol-3-O-β-D-glicopyranoside（5） and 3-O-β-authemisol（6）. The structure of compound 1 was elucidated by the analysis of IR, HRESI-MS, 1D and 2D NMR spectral data. Compounds 2--5 were isolated from Chlorophytum laxum R. Br. In addition, all the compounds were evaluated for cytotoxicity on the human nasopharyngeal carcinoma cancer cell line 5-8F. Among them, the newly identified 25-R-spirosta-3,5-dien-12β-ol（1） and diosgenin（2） exhibited high cytotoxicity on 5-8F cells, with IC50 values of 24.8 and 41.9 μmol/L, respectively.

铜胁迫下吊兰的生理响应及对铜的吸收特性

采用水培法研究了铜胁迫下吊兰的生理响应及对铜的吸收特性。实验结果表明:短期内低浓度的Cu 2+胁迫(15 mg/L)对吊兰的光合过程存在促进作用,高浓度的Cu 2+胁迫(30 mg/L和60 mg/L)对吊兰光合过程存在明显的抑制;随着Cu 2+胁迫浓度的增大,吊兰根和叶片组织中丙二醛的含量显著增加,并且叶片组织对Cu 2+胁迫作用的响应敏感性高于根系组织;吊兰根系组织中的铜含量水平均明显高于叶片组织,并且随着Cu 2+浓度的增大根组织中铜的含量水平也显著增高,而叶片组织中的铜含量水平则变化不明显。研究结论能为该植物应用于铜污染环境的修复提供理论参考。

盐胁迫对两种吊兰光合特性和气孔形态的影响

选用吊兰(Chlorophytum comosum)和金边吊兰(Chlorophytum comosum cv.variegatum)一年生幼苗进行盆栽试验,设置盐分(NaCl)梯度为0和0.8%,研究了盐胁迫对两种吊兰生长、光合特性及气孔形态的影响。结果表明,在盐胁迫下,两种吊兰的叶片叶绿素含量、净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr)均下降,而细胞间CO_2浓度(Ci)增加,说明非气孔限制因素可能是两种吊兰光合能力下降的主要原因。两种吊兰通过改变根冠比和气孔密度、开放度来适应盐胁迫环境,表现出了一定的耐盐性,但两个品种间的耐盐性并无明显差异。本试验过程中发现两种吊兰都有泌盐的现象,通过扫描电镜技术分析发现,两种吊兰叶片和幼茎表皮气孔内都有残留的盐颗粒存在,说明吊兰气孔不仅仅用于气体和水分的交换,可能还兼有泌盐的功能。

镉胁迫对吊兰生长与土壤酶活性的影响

选用观赏植物吊兰进行盆栽试验,研究了吊兰对Cd的耐性、高浓度Cd胁迫对土壤酶活性以及土壤有效态Cd含量的影响.结果表明,随着Cd浓度的不断提高,脲酶活性显著下降.过氧化氢酶和蔗糖酶活性在Cd浓度为10 mg.kg-1的时候均达到了顶峰,而土壤磷酸酶活性则在Cd浓度为50 mg.kg-1的时候最大.4种土壤酶对重金属的敏感顺序为:脲酶>磷酸酶>蔗糖酶>过氧化氢酶.吊兰对Cd具有很强的耐性,在1500 mg.kg-1Cd胁迫浓度范围内,吊兰对Cd的耐性指数均大于50%.土壤有效态Cd与土壤Cd添加量和土壤酶活性呈显著相关性,可将土壤有效态Cd含量和土壤酶活性这两类指标作为镉污染土壤的评价指标.

干旱胁迫对金心吊兰叶片活性氧及其清除系统的影响

以金心吊兰叶片边缘的绿色区域与中心黄白色区域为材料,研究了干旱胁迫对其活性氧及其清除系统的影响,结果表明,1)随着胁迫时间的延长,叶片各区域的过氧化氢(H2O2)、超氧阴离子(O2-·)含量和丙二醛(MDA)含量均表现出逐渐上升的趋势,而超氧化物歧化酶(SOD)与过氧化物酶(POD)活性则先上升后下降。2)随着胁迫浓度的增加,叶片中活性氧(H2O2,O2-·)含量、抗氧化酶(SOD,POD)活性和MDA含量表现出上升的趋势。3)采用隶属函数法对吊兰叶片的抗旱性进行了综合评价,其抗旱性表现出位置效应,叶片中心区域抗旱性强于边缘区域。4)吊兰叶片的抗旱性与叶绿素含量呈负相关,叶绿素含量相对高的区域其抗旱性相对较低,金心吊兰抗旱性强于野生型。

吊兰生长对锌污染土壤微生物数量及土壤酶活性的影响

选用观赏植物吊兰进行盆栽试验,通过测定吊兰根际、非根际以及未栽培吊兰的空白组土壤微生物数量、土壤酶活性及化学性质,研究吊兰对重金属锌污染土壤的修复作用。结果表明:土壤微生物数量、土壤酶活性、有机质含量及土壤呼吸作用强度均表现为吊兰根际组>吊兰非根际组>未栽培吊兰的空白组。而土壤锌总量、pH、电导率及氧化还原电位均表现为未栽培吊兰的空白组>吊兰非根际组>吊兰根际组。土壤锌浓度为200mg/kg时,细菌、真菌的数量最多,土壤呼吸作用强度、脲酶及磷酸酶的活性也达到最高值;土壤锌浓度为500mg/kg时,放线菌数量最多,且蔗糖酶活性达到顶峰。三类微生物对锌的敏感性顺序为真菌>放线菌>细菌。土壤酶对重金属锌的敏感性顺序为蔗糖酶>脲酶>磷酸酶>过氧化氢酶。通过微生物数量与土壤酶活性的双变量相关性分析可知,放线菌的变化对土壤酶的影响最大,其次是真菌,细菌影响最小。吊兰生长能够有效改善土壤环境,在重金属锌污染修复方面有广阔的应用前景。

3种百合科植物挥发物成分分析

为了探究百合科Liliaceae植物挥发物(VOCs)成分组成,采用活体植物动态顶空套袋采集法收集吊兰Chloro-phytum comosum,一叶兰Aspidistra elatior和文竹Asparagus plumosus 3种百合科植物VOCs,利用热脱附-气相色谱/质谱联用法分析了这3种植物VOCs的主要成分。结果表明:吊兰释放34种VOCs,包含萜类、酯类、醇类、醛类、酸类等5类化合物,其中二氢香茅醇相对含量最高(28.79%);一叶兰释放VOCs共鉴定出25种,包含醛类、酯类、醇类、萜类、酮类、烃类等6类化合物,其中壬醛(相对含量为12.12%)为其主要成分;文竹释放出38种VOCS,包含萜类醛类、酯类、醇类、醛类等4种化合物,主成分为牻牛儿醇(相对含量为27.90%)。3种百合科植物挥发物中都含有萜类化合物、醇类化合物、醛类化合物、酯类化合物。研究结果为百合科植物的综合开发利用提供了理论依据。

吊兰生长对土壤镉形态分布与含量的影响

选用观赏植物吊兰进行盆栽试验,研究吊兰对Cd的吸收累积情况以及吊兰生长对土壤Cd的化学形态特征及其含量的影响。结果表明,吊兰对Cd具有极强的累积能力,在土壤Cd浓度为10 mg/kg时,吊兰根和地上部分的Cd累积浓度分别为179.6,149.3 mg/kg,达到了超富集植物的标准。吊兰的生长通过促进残留态(RES)Cd向弱结合态Cd的转化,改变Cd的形态分布,活化土壤Cd,影响Cd的有效性,加大了重金属Cd的淋溶作用和植物对Cd的吸收累积,使实验组Cd含量显著降低(p<0.01),与空白组相比下降了13.71%。吊兰的生长降低了土壤Cd的污染程度,有利于Cd污染土壤的修复。