Exploring the mechanism of action bitter melon in the treatment ofbreast cancer by network pharmacology

BACKGROUND Bitter melon has been used to stop the growth of breast cancer(BRCA)cells.However,the underlying mechanism is still unclear.AIM To predict the therapeutic effect of bitter melon against BRCA using network pharmacology and to explore the underlying pharmacological mechanisms.METHODS The active ingredients of bitter melon and the related protein targets were taken from the Indian Medicinal Plants,Phytochemistry and Therapeutics and SuperPred databases,respectively.The GeneCards database has been searched for BRCA-related targets.Through an intersection of the drug’s targets and the disease’s objectives,prospective bitter melon anti-BRCA targets were discovered.Gene ontology and kyoto encyclopedia of genes and genomes enrichment analyses were carried out to comprehend the biological roles of the target proteins.The binding relationship between bitter melon’s active ingredients and the suggested target proteins was verified using molecular docking techniques.RESULTS Three key substances,momordicoside K,kaempferol,and quercetin,were identified as being important in mediating the putative anti-BRCA effects of bitter melon through the active ingredient-anti-BRCA target network study.Heat shock protein 90 AA,proto-oncogene tyrosine-protein kinase,and signal transducer and activator of transcription 3 were found to be the top three proteins in the proteinprotein interaction network study.The several pathways implicated in the anti-BRCA strategy for an active component include phosphatidylinositol 3-kinase/protein kinase B signaling,transcriptional dysregulation,axon guidance,calcium signaling,focal adhesion,janus kinase-signal transducer and activator of transcription signaling,cyclic adenosine monophosphate signaling,mammalian CONCLUSION Overall,the integration of network pharmacology,molecular docking,and functional enrichment analyses shed light on potential mechanisms underlying bitter melon’s ability to fight BRCA,implicating active ingredients and protein targets,as well as highlighting the major signaling pathways that may be altered by this natural product for therapeutic benefit.

Field Evaluation on Agronomic Characteristics of Newly Introduced Bitter Gourd(Momordica charantia L.)Accessions in Thua Thien Hue Province,Vietnam

The main objective of this study was to evaluate ability of growth and yield of introduced bitter gourd accessions in winter-spring 2016-2017 in Thua Thien Hue province.A total of seven accessions were used in this study.Of these,six accessions were provided by the World Vegetable Center(AVRDC),namely,AVRDC 1329,AVRDC 1330,AVRDC 1331,AVRDC 1333,AVRDC 1334,AVRDC 1335 and one Vietnam local accession(control)from Dien Hai commune,Phong Dien district,Thua Thien Hue province.The results showed that experiment accessions can grow under Thua Thien Hue conditions.AVRDC 1329,AVRDC 1330,and AVRDC 1335 were considered as displaying good growth and development ability.Of those,AVRDC 1330 was the most suitable to consumer as regards to appearance and bitter taste.AVRDC 1330,the control check and AVRDC 1331 had the high actual yield with 16.57,10.65 and 7.88 tons/ha,respectively,and these two introduced accessions can be used for breeding and cultivation under local condition.

The Use of HPLC in Determination of Endogenous Hormones in Anthers of Bitter Melon

A simple, rapid and perfect extraction and determination method for the endogenous hormones in anthers of bitter melon with reversed-phase high performance liquid chromatography （HPLC） has been developed. The HPLC system consisted of Hypersil ODS C 18 reverse phase column （150 mm × 4.6 mm, 5 μm） with a methanol gradient in 0.6% acetic acid and UV detector set at 254 nm. The results showed that the method was accurate and efficient.

Effects of Light-Emitting Diode (LED) Red and Blue Light on the Growth and Photosynthetic Characteristics of <i>Momordica charantia</i>L.

With andromonoecious Momordica charantia L. (bitter gourd) as material, three light qualities (50 μmol·m-2·s-1) including white LED light (WL), blue monochromatic light (B, 465 nm), and red monochromatic light (R, 650 nm) were carried out to investigate their effects on seed germination, physiological and biochemical parameters, sex differentiation and photosynthetic characteristics of bitter gourd. The results showed that compared to the WL treatment, the R treatment significantly promoted seed germination, seedling height elongation and soluble sugar content, the B treatment significantly increased seedling stem diameter, reducing sugar content and soluble protein content, the R and B treatments both significantly reduced sucrose content, but their POD activity showed no significant difference. Compared with the R treatment, the B treatment significantly increased the total female flower number and female flower nod ratio in 30 nods of main stems. The study of photosynthetic characteristics found that the R and B treatments could effectively increase the stomatal conductance (GS) of leaves, significantly improved the net photosynthetic rate (Pn) compared to the WL treatment, and the effect of the B treatment was better. Compared to the R and WL treatments, the B treatment increased the maximum photosynthetic rate (Pmax), apparent quantum efficiency (AQE) and light saturation point (LSP), and reduced the dark respiration rate (Rd) and light compensation point (LCP) of the leaves. Fit light response curves showed that the adaptability and utilization of weak light in bitter gourd were middle or below, but it showed higher adaptability and utilization of strong light. Thus, it suggests that Momordica charantia is a typical sun plan with lower Rd. In summary, it is concluded that blue light has a positive effect on the seed germination, seedling growth, sex differentiation and improving the photosynthetic performance, and this will lay the foundation for artificially regulating optimum photosynthesis using specific LEDs wavelength, and help to elucidate the relationship how light quality influences the sex differentiation of plant.

苦瓜种质资源耐热性评价及其鉴定指标的筛选

本研究以27个苦瓜自交系4叶1心的幼苗为材料,预培养1 d后,置于39℃高温培养,观察其高温胁迫72 h后的热害情况,通过计算热害指数(TDG)筛选评价出高、中、低耐热性的苦瓜种质;从中选取6个不同耐热性的自交系,测定39℃胁迫0 h和48 h的株高(PH)、茎粗(SD)、鲜物质量(SFW)和干物质量(SDW)等形态学指标以及叶片相对叶绿素含量(SPAD)、电导率(REL)、过氧化物酶活性(POD)、过氧化氢酶活性(CAT)、超氧化物歧化酶活性(SOD)、丙二醛含量(MDA)、脯氨酸含量(Pro)和可溶性糖含量(SS)等生理指标;主成分分析上述12个指标的耐热系数(α),综合评价6个自交系材料的耐热性;利用逐步回归分析法建立耐热性评价数学模型获得苦瓜耐热性鉴定指标体系。结果表明:以TDG为依据,27个苦瓜自交系中有耐热型3个,中等耐热型18个,热敏型6个;6个不同耐热性材料的综合评价结果与热害指数鉴定结果基本一致;耐热性的评价数学模型为:Y=-0.426+0.049X1-0.011X2-0.009X3-0.018X4,利用建立的最优回归方程预测供试材料的耐热性,预测值与Y值基本一致。说明SFW(X1)、POD活性(X2)、CAT活性(X3)和SS含量(X4)这4个指标的耐热系数可以快速准确地评价苦瓜耐热性差异,可用作苦瓜耐热性鉴定指标。本研究筛选出苦瓜耐热种质并构建苦瓜耐热性评价体系,为海南夏季耐热苦瓜新品种选育提供依据。

苦瓜枯萎病生防细菌ZB36的分离鉴定及其定殖特性研究

【目的】筛选并鉴定对苦瓜枯萎病具有稳定拮抗活性的生防菌株,明确其在苦瓜植株上的定殖特性。【方法】从山东省淄博市耕作地采集土壤样品,以苦瓜枯萎病菌为靶标,通过梯度稀释涂布平板法分离、纯化,获得1株具有稳定拮抗活性的纯培养细菌,命名为ZB36。根据菌株形态学特征、生理生化特性,以及16S rRNA基因和gyrA保守基因序列分析进行菌株鉴定。通过抗生素抗性标记法获得遗传特性和拮抗活性稳定的利福平标记菌株ZB36^(R),并对其在苦瓜上的定殖特性进行探究。【结果】菌株鉴定结果表明,ZB36为贝莱斯芽孢杆菌(Ba-cillus velezensis)。盆栽试验结果表明,ZB36不仅对苦瓜枯萎病具有显著的防治效果,防效高达74.76%,而且对苦瓜植株也具有显著促生作用,接种ZB36后苦瓜地上部鲜重和株高分别提高了28.65%和22.48%。定殖试验结果表明,ZB36在苦瓜根、茎和叶中均能定殖,且在接种后28 d仍能在这些组织中稳定定殖。【结论】贝莱斯芽孢杆菌ZB36不仅对苦瓜枯萎病具有良好的防治和促生效果,同时也具有较强的定殖能力,具备作为苦瓜枯萎病生防细菌资源开发利用的潜力。

苦瓜品质性状主基因+多基因混合遗传效应分析

开展苦瓜(Momordica charantia L.)干物质含量、可溶性蛋白质含量和硝酸盐含量的品质性状研究,分析其遗传机制。以苦瓜雌性系Z-1-4为母本,优质苦瓜高世代自交系2538、5325和5321为父本,构建3个组合的4个世代[P_(1)(亲本)、P_(2)、F_(1)、F_(2)]遗传群体,并结合主基因+多基因混合遗传模型对苦瓜干物质含量、可溶性蛋白质含量和硝酸盐含量进行遗传及相关分析。结果表明,苦瓜干物质含量最适遗传模型为等加性主基因加性+多基因模型,且遗传率较高;苦瓜可溶性蛋白质含量最适遗传模型为负向完全显性主基因+加性-显性多基因模型,遗传率较低,适合在早期世代进行遗传选择。商品瓜硝酸盐含量最适遗传模型为2对加性-显性主基因+加性-显性多基因模型,遗传率也较低。可溶性蛋白质与硝酸盐的遗传率均相对不高,适合在高世代进行表型选择。

Diverse roles of bitter melon(Momordica charantia)in prevention of oral cancer

Oral squamous cell carcinoma(OSCC)is one of the common lethal malignancies which is increasing rapidly in the world.Increasing risks from alcohol and tobacco habits,lack of early detection markers,lack of effective chemotherapeutic agents,recurrence and distant metastasis make the disease more complicated to manage.Laboratory-based studies and epidemiological studies indicate important roles of nutraceuticals to manage different cancers.The plant bitter melon(Momordica charantia)is a good source of nutrients and bio-active phytochemicals such as triterpenoids,triterpene glycosides,phenolic acids,flavonoids,lectins,sterols and proteins.The plant is widely grown in Asia,Africa,and South America.Bitter melon has traditionally been used as a folk medicine and Ayurvedic medicine in Asian culture to treat diseases such as diabetes,since ancient times.The crude extract and some of the isolated pure compounds of bitter melon show potential anticancer effects against different cancers.In this review,we shed light on its effect on OSCC.Bitter melon extract has been found to inhibit cell proliferation and metabolism,induce cell death and enhance the immune defense system in the prevention of OSCC in vitro and in vivo.Thus,bitter melon may be used as an attractive chemopreventive agent in progression towards OSCC clinical study.

苦瓜皂苷对饲喂高糖饲料松浦镜鲤幼鱼生长、消化及肝脏健康的影响

为探讨苦瓜皂苷对饲喂高糖饲料松浦镜鲤(Cyprinus carpio Songpu)幼鱼的生长、消化及肝脏健康的影响,随机选取450尾体质量(6.43±0.02)g的松浦镜鲤幼鱼饲养在室内500 L控温循环水族箱中,分别投喂蛋白30%、脂肪6%而含糖不同的6种饲料:含糖10%的饲料(对照组LC)和含30%饲料(高糖组HC)及含30%糖的高糖中添加100 mg/kg(MCS1)、400 mg/kg(MCS2)、1600 mg/kg(MCS3)及6400 mg/kg(MCS4)]苦瓜皂苷的饲料,在水温28℃~32℃下饲养8周。结果表明,MCS3组松浦镜鲤的终末体质量、特定生长率、饲料效率及摄食率显著高于HC组(P<0.05),且MCS3组的终末体质量和摄食率显著高于LC组(P<0.05)。MCS3组松浦镜鲤的蛋白酶及淀粉酶活性最高,显著高于HC组和MCS4组(P<0.05),而其脂肪酶活性最低,显著低于HC组(P<0.05)。MCS3组肝脏的核因子E2相关因子2、铜锌超氧化物歧化酶、锰超氧化物歧化酶及过氧化氢酶的表达量,超氧化物歧化酶、过氧化氢酶、过氧化物酶、谷胱甘肽、总抗氧化能力活性最高,显著高于HC组(P<0.05)。MCS3组Kelch样环氧氯丙烷相关蛋白1表达量和丙二醛含量最低,显著低于HC组(P<0.05);MCS3组中过氧化氢酶表达量显著高于LC组(P<0.05)。MCS3组改变了HC组的病理表现,肝细胞核更清晰,细胞界限明显且排列整齐,与LC组肝脏组织结构完整度相似,结构较清晰。综上所述,高糖饲料中苦瓜皂苷添加水平应在1600 mg/kg左右,当添加水平达到6400 mg/kg时,不利于松浦镜鲤幼鱼肠道消化及肝脏健康。

不同果肉厚度的苦瓜种质细胞大小和形态差异分析

为探讨苦瓜果肉生长规律及细胞大小和形态对果肉发育的影响,以2份不同果肉厚度苦瓜种质(分别为LX13、ZK54)为材料,分析不同时期果肉厚度动态变化以及细胞大小和形态参数,研究不同指标之间的相关性。结果表明:(1)2份苦瓜种质果肉厚度随果实发育不断增大,在花后17、23 d种质间差异显著。(2)LX13细胞面积在花后10、17和23 d差异显著,细胞周长、长度、宽度在4个取样时间差异显著,ZK54细胞面积花后17 d显著大于花后10 d,细胞周长、长度在花后3、10和17 d差异显著,细胞宽度在4个时间差异显著;在花后17、23 d,LX13细胞面积、细胞周长、长度和宽度均显著大于ZK54。(3)LX13细胞纵横比和圆度在花后17、23 d显著大于花后3、10 d,ZK54两个指标在花后17 d显著大于花后3、10 d,但花后23 d这两个指标均显著小于花后17 d;在花后17、23 d,LX13的细胞纵横比和圆度显著大于ZK54。(4)相关分析显示细胞参数与果肉厚度之间均呈显著或极显著正相关,表明细胞发育对苦瓜果实的生长起关键作用。

我国苦瓜新品种选育与保护现状分析

通过分析自2013年苦瓜纳入《中华人民共和国农业植物品种保护名录》之后文献报道的60个苦瓜品种,发现苦瓜瓜形以珍珠瓜为主,主要育种目标包括抗性、产量和成熟期,选育方式以杂种优势育种为主。截至2022年8月,我国苦瓜新品种权申请量130件,其中文献报道的60个品种中仅5个申请了新品种权;新品种权授权量30件,繁殖材料提交不及时或不合格是影响授权审查速度的主要原因,通过提前委托DUS测试能大幅缩短授权时间。

52份苦瓜种质种子低温发芽能力鉴定与评价

为评价苦瓜种子的低温发芽能力,测定了52份苦瓜种质种子在15℃处理下的8个发芽指标,并运用因子分析、相关分析、主成分分析、隶属函数分析、聚类分析、回归分析等方法进行综合性评价。结果表明:(1)52份苦瓜种质种子的发芽率、发芽势、生长势、发芽指数、活力指数、平均发芽时间、平均发芽速度和种子萌发指数均出现不同程度的差异,变异幅度为25.648%~127.182%。(2)发芽率、发芽势、生长势、发芽指数、活力指数、种子萌发指数两两之间呈极显著正相关,平均发芽速度与其他所有的指标均呈极显著负相关。(3)主成分分析共提取到两个主成分,累积贡献率为87.773%,其中主成分1贡献率为71.313%,主成分2贡献率为16.460%。(4)聚类分析将参试材料分成4个类型,基于综合评价值排序发现BG9、BG5、BG15、BG8和BG55的低温发芽能力强。(5)建立低温发芽能力评价方程D=0.162+0.001×X_(1)+0.003×X_(2)+0.137×X_(3)+0.02×X_(5),为挖掘及培育耐冷品种提供科学依据,也为简化生产流程、提高生产效率提供理论参考。

苦瓜籽核糖体失活蛋白的基本性质研究

采用 Affi- Gel亲和层析和 Sephacryl S- 10 0分子筛层析等方法获得苦瓜籽核糖体失活蛋白 (RIP) ,经 SDS- PAGE,PAGE,IEF和 PAS方法分析均表明为单一蛋白着色带或单一糖蛋白着色带 .根据 SDS- PAGE测得表观分子量为 30 0 0 0 ,经 IEF- PAGE结果计算其 PI为 9.0 ,对无细胞系统中蛋白质生物合成抑制活性明显 ,其 IC50 为 5.3× 10 -10 mol/ L左右 .体外生物活性试验结果表明其对人肝癌细胞 ,Vero,SP2 / 0 ,3T3,Kb,Navana,S- 180等肿瘤细胞株和麻疹病毒均表现有不同程度的抑制作用 ,而对完整人胚肺二倍体细胞却毒性极小 .

苦瓜籽核糖体失活蛋白的理化性质及生物活性

采用硫酸铵分级分离，假配基亲和层析和ＳｅｐｈａｃｒｙｌＳ－１００分子筛层析等方法，从苦瓜籽中获得核糖体失活蛋白（ＲＩＰ）．经ＳＤＳ－ＰＡＧＥ、ＰＡＧＥ、ＩＥＦ和ＰＡＳ方法分析均表明为单一蛋白着色带或单一糖蛋白着色带．根据ＳＤＳ－ＰＡＧＥ和Ｓｅｐｈａｄｅｘ Ｇ－１５０分子筛层析结果计算其相对分子质量为３．０×１０４，经ＩＥＦ－ＰＡＧＥ结果计算其ｐＩ为８．９～９．０．对无细胞系统中蛋白质生物合成抑制活性明显，其ＩＣ５０为５．３×１０－ １０ ｍ ｏｌ／Ｌ左右．体外生物活性试验结果表明其对人肝癌细胞、Ｖｅｒｏ、ＳＰ２／０、３Ｔ３、Ｋｂ、Ｎａｖａｎａ 等肿瘤细胞株均表现有不同程度的抑制作用．而对完整细胞人胚肺二倍体细胞却毒性极小．因此，上述实验结果为该ＲＩＰ的进一步深入研究和有可能开发成免疫毒素的高效弹头药物提供了一定的工作基础．

苦瓜单瓜种子数与种皮颜色的遗传分析

以苦瓜自交系29和惠州大顶为亲本构建6个世代(P1、P2、F1、B1、B2、F2)群体,分别对苦瓜单瓜种子数和苦瓜种皮颜色进行遗传分析。结果表明,苦瓜单瓜种子数的遗传符合2对等显性主基因+加性-显性多基因混合遗传模型(E-6模型),由主基因和多基因共同控制,多基因显性效应较大,F1表现出超亲优势。苦瓜种皮颜色的黑色对棕黄色表现为1对基因控制的显性遗传。

苦瓜种质遗传多样性的RAPD和ISSR分析

采用RAPD和ISSR分子标记技术对38份苦瓜种质进行遗传多样性分析。结果表明：10个RAPD和10个ISSR51物分别扩增出93条和81条带，多态性比率分别为50．54％和61．29％；RAPD和ISSR标记检测供试材料的遗传相似性系数（Gs）范围，分别为0．287～1和0．221～1，ISSR（平均GS值0．672）检测多态性效果高于RAPD（平均G5值0．694）。RAPD标记聚类分析将供试种质分为3个类群6组，分类结果与苦瓜瓜瘤的表型分类比较相似；ISSR标记聚类分析将供试种质分为3个类群7组，ISSR标记划分类群与形态上以颜色分类比较接近。RAPD和ISSR标记的遗传相似性系数呈显著相关（r=0．550）。两个标记整合后聚类分析可检测到更大的遗传变异，结果与苦瓜的农艺性状分类和地理分布有一定的相关性。

苦瓜抗白粉病SCAR分子标记的开发

【目的】获得与苦瓜抗白粉病基因紧密连锁的分子标记,为加快苦瓜抗白粉病新品种的选育奠定基础。【方法】以高抗白粉病野生苦瓜MC18为父本、高感白粉病苦瓜栽培种MC1-2为母本创建F2代分离群体;经单株抗病性鉴定后,以BSA法构建F2代单株的高抗和高感白粉病苦瓜DNA近等基因池;利用SRAP技术筛选多态扩增片段,对仅在抗白粉病近等基因池和父本中出现的差异片段进行回收、测序、比对和转化成SCAR标记,并利用已知抗病性的单株DNA分析标记与苦瓜抗白粉病的相关性。【结果】从1188对SRAP引物组合中筛选到稳定阳性差异条带的引物组合5对,其中ME20EM5引物对扩增的差异条带长度为332 bp,与葡萄抗体蛋白基因(抗性基因)的DNA序列有较高相似性,并将其成功转化成与苦瓜白粉病抗性相关、大小为320 bp的SCAR标记。【结论】开发的SCAR-ME20EM5分子标记可用于苦瓜抗白粉病分子标记辅助选择。

苦瓜主要农艺性状的相关及通径分析

对华南长身型苦瓜 14个主要农艺性状进行了相关及通径分析 ,相关分析结果表明 ,前期产量、延续收获期、结果数等 3个性状与产量呈极显著正相关 ,初花期、开花期、第 1雌花节位、第 1坐果节位等 4个性状与产量呈极显著负相关 ;通径分析结果表明 ,结果数、延续收获期和单瓜质量 3个性状对产量形成的直接作用最大 ,这 3个性状对产量的决定系数总和达 0 .910 8。

苦瓜种质资源遗传多样性的AFLP分析

对36份苦瓜种质资源进行AFLP-银染分子标记技术扩增,进行遗传多样性和亲缘关系评价分析。筛选8个多态性高、分辨力强的E+3/M+3引物组合对36份供试材料的基因组DNA扩增,共获得条带数1142条,多态性条带数992条,扩增效率87%,平均每对引物扩增条带数142.75条。其中,以E-ACA/M-CAG引物的扩增效率最高,达到91.79%;其次是E-AAG/M-CTA,达到90.30%;扩增效率最低的引物是E-ACA/M-CTT,为82.31%。表明供试材料在DNA水平上酶切位点的分布存在广泛的变异。各供试材料相似系数介于0.64～0.89;UPGMA分析将36份资源从变种间水平分为2个类群。

癞葡萄籽油脂肪酸组成的GC/MS分析

采用2-氨基-2-甲基丙醇(AMP)衍生化,GC/MS法分析癞葡萄籽油的脂肪酸组成。测定结果为:软脂酸1.65%,硬脂酸31.78%,油酸5.52%,亚油酸3.86%,十八碳共轭三烯酸55.13%,未定出成分2.07%。癞葡萄籽油的主要成分为十八碳脂肪酸,其中十八碳不饱和脂肪酸占64.51%。AMP衍生化GC/MS法可对不饱和脂肪酸双键的位置准确定位,将癞葡萄籽油中的3种共轭三烯酸的顺反异构体分开。