诺丽(Morinda citrifolia L.)离体快速繁殖研究

以诺丽（Morinda citrifolia L．）种子为试材．诺丽子叶和下胚轴均能单独由细胞分裂素BA0．7～2．0mg／L谤导不定芽发生，不定芽可直接从外植体发生，也可从愈伤组织发生，添加生长素NAA0．05～0．1mg／L则完全抑制不定芽发生，同时强烈促进愈伤组织和不定根发生．茎段在BA0．5～5．0mg／L或ZT 1．5mg／L或KN 1．5mg／L时均能通过腋芽萌发和不定芽发生而增殖．芽梢在NAA 0．1mg／L、IBA 0．1mg／L或IAA 0．1mg／L均有根群发生，但NAA 0．1mg／L诱导生根时切口愈伤组织较多，部分不定根由愈伤组织发生，而IAA 0．1mg／L诱导生根时根群欠发达，以IBA 0．1mg／L为最佳．试管苗移植到泥炭上10d后即可在全光照下生长．

海巴戟(Morinda citrifolia L.)组织培养研究

以海巴戟（Morinda citrifolia L．）种子为试材，在不剥除种皮的情况下，在MS无激素培养基上播种1年内未见发芽，在剥除种皮的情况下，在MS无激素培养基上发芽率最高，50d内可达75％。海巴戟子叶和下胚轴均能单独由细胞分裂素BA0．7～2．0mg／L诱导不定芽发生，不定芽可直接从外植体发生，也可从愈伤组织发生，添加生长素NAA0．05～0．1mg／L则完全抑制不定芽发生，同时强烈促进愈伤组织生长和不定根发生。带芽茎段在BA1．5mg／L配合低浓度生长素时均能通过腋芽萌发和不定芽发生而增殖。芽梢在NAA0．1mg／L、IBA0．1mg／L或IAA0．1mg／L均有根群发生，但NAA0．1mg／L诱导生根时切口愈伤组织较多，部分不定根由愈伤组织发生。而IAA0．1mg／L诱导生根时根群欠发达，以IBA0．1mg／L最佳。

诺丽(Morinda citrifolia Linn.)离体培养研究

为诺丽遗传转化奠定基础及诺丽工厂化育苗提供依据,本实验以诺丽茎尖及带芽茎段为外植体,以3%MS培养基添加不同的激素组合进行离体培养,选择最佳的培养组合。实验结果表明,茎尖在MS或添加NAA(0.3 mg/L)的MS培养基中均可得到完整植株,且在后者中的长势更好。该研究成功建立了诺丽离体再生体系,为规模化生产及遗传转化提供了技术支撑。

Cytoprotective effects of Morinda offucinalis against hydrogen peroxide-induced oxidative stress in Leydig TM3 cells

Aim： To investigate the antioxidant effects of Morinda officinalis （Morindae radix, MR） on H2O2-induced oxidative stress in cultured mouse TM3 Leydig cells. Methods： We carried out 2,2-diphenyl-1-picrylhydrazyl free radical scavenging, 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide, lipid peroxidation, testosterone enzyme immunoassay, superoxide dismutase （SOD）, and catalase （CAT） assays in Leydig TM3 cells. Results： MR showed a 47.8% 2,2-diphenyl-1-picrylhydrazyl radical scavenging effect in TM3 cells with no significant cytotoxicity. Oxidative stress was induced in TM3 cells with 100 μmol H2O2, and treatment of the cells with 250 μg/mL MR showed the most significant protective effect （64%, P 〈 0.001） in the cell viability assay with a decreased lipid peroxidation level （1.75 nmol/mg protein, P 〈 0.05）, increased testosterone production （43.5 pg/mL）, and improvements in SOD activity （7.49 units of SOD/mg protein, P 〈 0.001） and CAT activity （74.6 units of CAT/mg protein, P 〈 0.001）. Conclusion： These findings indicate that MR, as an antioxidant, protects functions of cultured mouse TM3 Leydig cells from H2O2-induced oxidative stress.

Antioxidant and cytotoxic effects of hexane extract of Morinda pubescens leaves in human liver cancer cell line

Objective:To evaluate the antioxidant and cytotoxic effects of hexane extract of Morinda pubescens leaves and to find the primary bioactive compound responsible for antioxidant and cytotoxic activities.Methods:The individual compounds were isolated using column chromatography and were characterized by spectroscopic techniques.The antioxidant activity was evaluated for all individual isolated compounds by DPPH method using L-Ascorbic acid as standard and cytotoxicity was assessed for the extract and the hyoscyamine by MTT assay, caspase test and RT-PCR study.Results:The antioxidant activity of the isolated compounds and the extract increased as the concentration increased.One of the isolated compound hyoscyamine showed the high antioxidant activity.The extract and the hyoscyamine dose-dependently decreased the cell viability in HepG2 cells.Hyoscyamine induced caspase-3 mediated apoptosis. Up regulation of p53 gene expression provides cue for apoptotic activity of hyoscyamine. Conclusions:The results indicate that hexane extract possessed potent antioxidant and cytotoxic activity and hyoscyamine is the principal bioactive compound in hexane extract.

Extraction and Characterization of Antioxidant Compositions From Fermented Fruit Juice of Morinda citrifolia (Noni)

Extraction and characterization of antioxidative compositions from the extracts of fermented Xisha Noni （Morinda citrifolia L.） juice were studied. The antioxidative constituents of 184.6 g freeze-dried extracts of naturally fermented Xisha Noni juice were isolated successfully by petroleum ether, EtOAc and n-BuOH solvents, and the antioxidative effects were measured according to scavenging activity against hydroxyl generated in Fenton reaction system and superoxide anion radicals in pyrogallol autoxidation system. The EtOAc extract exhibited most significantly higher （P 〈 0.01） antioxidative activity than mannitol or vitamin C, while the petroleum ether and n-BuOH extracts showed lower activities compared to mannitol. Three antioxidant phenolic compounds, isoscopoletin, aesculetin and 3,3＇,4＇,5,7-pentahydroxyflavone （quercetin） were isolated from the EtOAc extract by several chromatography techniques for the first time. The results suggest that several compounds, in particular, the phenolic compounds, contribute separately or synergistically to the antioxidative activity of fermented Noni fruit juice.

Antioxidant and antiangiogenic activities of the essential oils of Myristica fragrans and Morinda citrifolia

Objective:Toinvestigate the anti-angiogenic activity and antioxidant properties of Myristica fragrans（M.fragrans）（nutmeg） and Morinda citrifolia（M.citrifolia）（mengkudu） oils. Methods:The nutmeg and megkudu essential oils were obtained by steam distillation. The antioxidant activities of both essenlial oils were delermined by beta-carotene/ linoleic acid bleaching assay and reducing power while the anti-angiogenic activity was investigated using rat aortic ring assay using various concentrations.Results:The results showed that nutmeg oil has higher antioxidant activity than mengkudu oil.The nutmeg oil effectively inhibited the oxidation of linoleic acid with（88.68±0.1）%while the inhibition percentage of oxidation of linoleic acid of the mengkudu oil is（69.44±0.4）%.The nutmeg oil and mengkudu oil showed reducing power with an EC50 value of 181.4μg/mL and 3 043.0μg/mL,respectively.The anliangiogenic activity of nutmeg oil showed significant antiangiogenic activity with IC50 of 77.64μg/mL comparing to mengkudu oil which exhibits IC50 of 109.30μg/mL.Conclusion:Bioactive compound（s） will be isolated from the nutmeg essential oil to be developed as antiangiogenic drugs.

Antispermatogenic activity of Morinda lucida extract in male rats

Aim： To investigate the effect of Morinda lucida Benth （Rubiaceae） on the reproductive activity of male albino rats. Methods： Two groups of rats were treated with 400 mg/（kg.d） of Morinda lucida leaf extract for 4 and 13 weeks, respectively. The control rats received the vehicle. All the treated rats had corresponding recovery groups. At the end of each experimental period, animals were killed and organ weights, sperm characteristics, serum testosterone levels, histology of the testes and fertility were assessed. Results： Morinda lucida leaf extract did not cause any changes in body and somatic organ weights, but significantly increased the testis weight （P 〈 0.05）. The sperm motility and viability, and the epididymal sperm counts of rats treated for 13 weeks were significantly reduced （P 〈 0.05）. Sperm morphological abnormalities and serum testosterone levels were significantly increased （P 〈 0.05）. There were various degrees of damage to the seminiferous tubules. The extract reduced the fertility of the treated rats by reducing the litter size. Reversal of these changes, however, occurred after a period of time. Conclusion： The extract of Morinda lucida has reversible antispermatogenic properties.

Mechanistic action of the acute toxicity of Bajitian(Morinda officinalis)in zebrafish embryos

Objectives:To investigate acute toxicity of Bajitian(Morinda officinalis)in zebrafish embryos.Methods:Zebrafish embryos at 48-h post fertilization(hpf)were exposed to Bajitian ethanol extract for72 h.The causative action of a delay in yolk sac absorption by Bajitian was investigated by RT-PCR analysis of lipid metabolism-related microsomal triglyceride transfer protein(MTP),apolipoprotein CII(ApoC2)and lipogenesis-related liver x receptor(LXR)genes.The effect of Bajitian eliciting an inflammatory response was studied by exposing 72 hpf myeloperoxidase(MPO):GFP transgenic zebrafish embryos to Bajitian extract for 4 h.Assessment was done by TUNEL,caspase-3/7,and RT-PCR analysis of the apoptosis related pathway B-cell lymphoma 2 associated X protein(Bax),Nuclear factor kappa-lightchain-enhancer of activated B cells(NF-k B)genes,neutrophil development-related stem cell leukaemia(SCL)and transcription factor PU.1 genes,to reveal the causative action of Bajitian reducing neutrophils.Results:RT-PCR analysis found that Bajitian extract had no effect on the expression of MTP or ApoC2 genes,but upregulated LXR gene,which might explain the delay in yolk sac absorption.Analysis of the inflammatory response showed that compared with negative controls,Bajitian extract significantly(P<.05)reduced the number of neutrophils in MPO:GFP embryos.TUNEL,caspase-3/7,and RT-PCR analysis of Bax and NF-k B genes found that Bajitian extract did not trigger the cell apoptosis.Further RT-PCR analysis found that Bajitian extract did not affect SCL expression,but did lead to down-regulation of PU.1.The inhibition of neutrophil development/differentiation may explain the decline in the total number of neutrophils following Bajitian treatment,which could be attributed to the anti-inflammatory effects found clinically for this drug.Conclusions:This study demonstrated that Bajitian caused a delay in yolk sac absorption and a decrease neutrophil in zebrafish embryos,which may be related to the inhibition of neutrophil development.

Effect of <i>Morinda citrifolia</i>L. fruit juice on gingivitis/periodontitis

The majority of the world’s population suffers from gingivitis/periodontitis. This inflammatory process is caused by several bacterial species inside the dental plaque. In vivo and in vitro experiments were set up. The patients of the in vivo group were divided into a noni and a control group. Both groups contained patients that suffered from gingivitis/periodontitis who were introduced to practice standardized, good oral hygiene. The patients in the noni group additionally used noni juice for mouth wash two times a day. The Papillae-Bleeding-Index (PBI) was evaluated comparing the status of inflammation in both groups. Bacterial probes were isolated from the patient’s gingival pouches for species identification and to carry out in vitro experiments for possible antimicrobial effects of noni juice. The Papillae-Bleeding-Index (PBI) in the noni group has “highly significantly” improved from an average of 2.25 at the beginning of the observation period (t0) to 1.01 after four weeks of noni treatment (t1), compared to a change from 2.11 at t0 to 1.95 at t1 inthe control group. A comparison of the differences of the PBI-values (t0-t1) between the noni and the control group was highly significant using a t-test on a level of p = 0.01. Only small inhibition zones were observed in the agar diffusion test on agar plates coated with aerobic, anaerobic and Candida cultures isolated from the patients gingival pouches after treatment with original or neutralized noni juice in different concentrations. Weak bacteriostatic effects occurred in the agar dilution experiments with noni juice in higher concentrations (native and neutralized noni juice). The present investigation has shown that the combination of good oral hygiene and the administration of noni juice was a promising treatment for gingivitis and periodontitis. The additional treatment with noni juice significantly mitigated the gingival inflammation.

Analysis of Volatile Components in Fresh Fruit of Morinda citrifolia L.from Different Origins Based on HS-SPME-GC/MS

[Objectives]This study was conducted to clarify the differences of volatile components in fresh fruit of Morinda citrifolia L.from different origins.[Methods]The method of HS-SPME-GC/MS detection was used to determine the volatile chemical components in fresh fruit of M.citrifolia.[Results]52,52 and 45 volatile components were identified from the fresh fruit of M.citrifolia from Xisha,Wanning and Haikou,respectively.Among them,the number and content of the identified esters were relatively high,mainly methyl caprylate,ethyl caprylate,4-pentenyl caproate,3-methylbuten-2-enyl caprylate,methyl caproate,and ethyl caproate.There were also fatty acids,alcohols,phenols,ketones,aldehydes and other substances.There were 33 common ingredients in the fruit from the three origins,mainly including caprylic acid,caproic acid,capric acid,methyl caprylate,ethyl caprylate,4-pentenyl caproate,3-methylbutene-2-enyl caprylate,methyl caproate,ethyl caproate,methyl caprate and hexyl caproate.[Conclusions]The types and contents of volatile components in fresh fruit of M.citrifolia from different origins were significantly different.

Lignans from <i>Morinda citrifolia</i>(Noni) Fruit Inhibit Fatty Acid Amide Hydrolase and Monoacylglycerol Lipase <i>in Vitro</i>

Morinda citrifolia (noni) fruit juice has exhibited a variety of biological activities in human clinical trials, indicating that it influences multiple systems of the body. Since the 1990s, the endocannabinoid system (ECS) has been found to modulate the activity of other organ systems. To investigate noni’s potential impact on the ECS, extracts from freeze-dried noni fruit were evaluated in fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) inhibition assays. The ethyl acetate extract demonstrated the greatest activity against both enzymes. Lignans in this extract also inhibited enzyme activities, with americanin A being the most active in both assays. Americanoic acid and 3,3’-bisdemethylpinoresinol were the next most active compounds. These results suggest that lignans in noni fruit may influence endocannabinoid levels within the body via FAAH and MAGL inhibition. This reveals another set of probable mechanisms of action by which noni juice affects human health.

<i>Morinda citrifolia</i>(Noni) Fruit Juice Inhibits Endocannabinoid Degradation Enzymes

Morinda citrifolia (noni) fruit juice has been shown to have a wide variety of potential health benefits in human clinical trials. It may also influence the endocannabinoid system of the body. Since the main ingredient of the product studied in these clinical trials was juice made from noni fruit puree from French Polynesia, it was evaluated for its ability to inhibit the two major endocannabinoid degradation enzymes in vitro. Noni fruit juice inhibited both fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) in a concentration-dependent manner, suggesting that it may help maintain anandamide and 2-arachidonoylglycerol levels. Samples of the puree were also analyzed for the presence of characteristic phytochemical markers of authentic noni fruit such as scopoletin, rutin, quercetin, deacetylasperulosidic acid and asperulosidic acid, all of which were present. Also present was scandoside, which is reported for the first time as being identified in noni fruit or its juice. Some of these compounds may contribute to the FAAH and MAGL inhibiting activity of noni juice. These results reveal another set of mechanisms by which noni juice possibly supports mental health, maintains joint health, relieves discomfort and modulates the immune system.

Mineral and Trace Element Concentrations in <i>Morinda citrifolia</i>L. (Noni) Leaf, Fruit and Fruit Juice

Fruit and leaf of noni (Morinda citrifolia L.) have been used traditionally as food and for medicinal purposes by South Pacific populations since over two thousand years. Recently noni fruit juice became very popular as health beverage worldwide. Manufacturers and distributors of noni juice often praise the high content of minerals and trace elements in conjunction with the geological situation of the origin of the raw noni material. We therefore performed an investigation about the metal concentration of noni fruit, leaf and soil samples from 42 different locations in French Polynesia, and of 16 commercial noni juices. Median concentrations of metals in noni fruit, leaf and fruit juice appeared in a normal range compared to other fruits, however, the variance between different locations was considerable. Noni fruits from lime soil were poorer compared to volcanic soil in most metals. Self-prepared noni juices did generally not serve for more then 10% of the daily requirement of trace elements and minerals per 100 ml. The metal concentration in noni fruit puree was higher compared to clear juice and reached 10% and 34% of the daily requirement in 100 ml for manganese and chromium respectively. The concentrations of toxic metals (As, Cd, Pb) were below 2% of the maximum permitted levels (MPL) in all of these juices. For one of the commercial noni juices the MPL for lead was exceeded (170%) and some others were close to it. Most metals showed a good correlation regarding the concentration in noni fruit versus leaf, but not for soil versus fruit or juice. The concentration of magnesium, manganese, zinc and germanium in the commercial juices was highly correlated to the potassium concentration, indicating that these elements are useful to detect a dilution of noni juices.

The Influence of <i>Morinda citrifolia</i>(Noni) Fruit Juice on Collagen Deposition in the Skin: A Minireview

The fruit of Morinda citrifolia, commonly known as noni, has an extensive history of use as a food and medicine throughout the tropics. Among its many uses, noni was believed to promote skin health. In vivo wound healing studies reveal that noni fruit juice ingestion increases collagen production and deposition via increased expression of extracellular matrix protein genes. Noni juice also appears to promote fibroblast proliferation. In vitro studies indicate that noni juice also protects fibroblasts via activation of the transcription factor Nrf2 and protects extracellular matrix collagen by inhibiting matrix metalloproteinases. In vitro and in vivo study findings are corroborated by a clinical trial where hospitalization times were reduced, and wound granulation quality was improved in pediatric burn patients. The findings of other human studies also indicate that noni juice has the potential to protect skin collagen. These include reduced plasma reactive oxygen species in cigarette smokers, lowered skin glycation levels in overweight and obese adults, as well as reduced skin glycation levels among noni juice consumers within the general population. These in vitro, in vivo and human studies reveal that there are multiple mechanisms of action through which noni juice promotes collagen synthesis and inhibits collagen degradation. The procollagen properties of noni juice aid wound healing, increase skin elasticity, and improve the overall appearance of the skin.

Effects of Moringa oleifera and Morinda citrifolia Extracts on Performance of Multiparous Sows and Suckling Piglets

[Objective]The test was to investigate the effects of MM(Moringa oleifera and Morinda citrifolia)extracts on reproductive performance of multiparous sows and growth performance of suckling piglets.[Method]A total of 60 multiparous sows(Landrace×Large White,2-4 parities)were randomly assigned into two treatments,30 replicates per treatment and one sow per replicate.Sows in control group were fed with basal diet,and those in experimental groups were fed with the basal diet supplemented with 500 mg/kg MM extract.The trial lasted 28 d(from 7 d before delivery to 21 d post lactation).[Result]MM extract had no significant effect on sow’s stage of labor and estrus rate at 7 d post weaning(P>0.05).MM extract had no significant impact on growth performance of suckling piglets(P>0.05).[Conclusion]Addition of MM extracts in the diet of multiparous sows at 7 d before delivery did not improve the performance of sows and piglets,but shortened sow’s stage of labor and promoted estrus of sows after weaning to a certain extent.

Study on the Extraction Process of Total Flavonoids from Morinda citrifolia L.

[Objectives]This study was conducted to explore the best extraction technology of total flavonoids from Morinda citrifolia L.[Methods]An orthogonal test was carried out to select the best extraction process with the yield of total flavonoids in the extract as the investigation index.[Results]Among the four factors of experimental design,the extraction time had a very significant impact on the results,the ethanol concentration had a significant impact on the results,and the extraction temperature and ethanol dosage had no significant impacts on the results.The optimal extraction process conditions for the total flavonoids of M.citrifolia were as follows:extraction time 1.5 h,ethanol concentration 90%,extraction temperature 60-65℃,and ethanol dosage 1∶20(g/ml).The total flavonoids in M.citrifolia fruit were extracted according to the optimal extraction process conditions,and the yield was 2.936%.[Conclusions]This study provides a theoretical basis for obtaining the optimal extraction process conditions for total flavonoids of M.citrifolia.

<i>Morinda citrifolia</i>(Noni) Fruit Juice Inhibits SARS-CoV-2 Spike Protein Binding of Angiotensin-Converting Enzyme 2 (ACE2)

COVID-19 is a global pandemic that has claimed millions of lives. This disease is caused by a coronavirus, SARS-CoV-2, which requires the binding of its spike protein to angiotensin-converting enzyme 2 (ACE2) for infection of the host cell. Morinda citrifolia (noni) fruit juice has antiviral activity that involves enhancement of immune system function. SARS-CoV-2 spike-ACE2 interaction experiments were carried out to further investigate the antiviral properties of noni juice and its major iridoids. Noni juice inhibited binding by approximately 69%. Scandoside was the most active of the three iridoids evaluated, reducing average spike protein-ACE2 interaction by 79.25%. The iridoids worked synergistically towards inhibiting spike protein binding when assayed together, improving activity by more than 22% above the expected level. But the modest activity of the most abundant iridoid, deacetylasperulosidic acid, indicates that other phytochemicals (i.e. scopoletin, quercetin, rutin and kaempferol) are also involved. Our results suggest that the presence of several biological active phytochemicals in noni juice enhances resistance to SARS-CoV-2 by interfering with its ability to bind ACE2. This is a new and significant anti-viral mechanism of noni juice that does not directly involve its immunomodulatory properties.

不同产地巴戟天HPLC指纹图谱及化学模式识别研究

目的:利用HPLC指纹图谱及化学模式识别对巴戟天药材进行评价。方法:采用Shodex Asahipak NH2P-504E色谱柱(4.6 mm×250 mm,5μm),以乙腈-水为流动相梯度洗脱,使用蒸发光散射检测器(ELSD)。利用相似度评价、聚类分析和主成分分析等化学模式识别方法,对不同产地的巴戟天药材进行分析。结果:所建立的HPLC指纹图谱共识别了18个共有峰,经与对照品图谱比较,鉴定其中的4个共有峰分别为果糖、蔗糖、蔗果三糖和耐斯糖。6个产地巴戟天的相似度在0.861~0.995;经聚类分析和主成分分析,6个产地巴戟天共聚为3类。结论:本研究采用的巴戟天HPLC指纹图谱及化学模式识别技术可反映出不同产地的巴戟天样品的差异,可为巴戟天的质量控制提供参考。