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18 pages, 8313 KB  
Article
Integrating Network Pharmacology and Experimental Validation: Mechanistic Study of the Anti-Porphyromonas gingivalis and Anti-Inflammatory Effects of Berberis hemsleyana Ahrendt Extract
by Guibin Yang, Mingan Gui, Hai Dong, Dongzhi Zhuoma, Xuehuan Li, Tai Shen, Hao Guo, Ruiying Yuan and Le Li
Plants 2026, 15(1), 115; https://doi.org/10.3390/plants15010115 - 31 Dec 2025
Cited by 1 | Viewed by 671
Abstract
Anti-Porphyromonas gingivalis mechanisms of Berberis hemsleyana bark extract remain to be elucidated, and the anti-inflammatory activity of its n-butanol fraction (BNB) in RAW264.7 cells—mediated through suppression of the NF-κB pathway—require further validation. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) [...] Read more.
Anti-Porphyromonas gingivalis mechanisms of Berberis hemsleyana bark extract remain to be elucidated, and the anti-inflammatory activity of its n-butanol fraction (BNB) in RAW264.7 cells—mediated through suppression of the NF-κB pathway—require further validation. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the crude extract from B. hemsleyana were determined against Candida albicans, Escherichia coli, Porphyromonas gingivalis, Staphylococcus aureus and Streptococcus mutans. Scanning electron microscopy (SEM) and bacterial protein leakage assays were used to evaluate its antibacterial activity against P. gingivalis. High-performance liquid chromatography-mass spectrometry (LC-MS) was applied to analyze the ethanol extract of B. hemsleyana bark, leading to the screening of 47 compounds. The antibacterial mechanisms of the compounds were predicted through Network Pharmacology analysis and Molecular docking. Anti-inflammatory activity mediated via the NF-κB pathway was verified using an LPS-induced RAW264.7 cell inflammatory model. Specifically, the BNB showed a significant antibacterial effect on P. gingivalis. Meanwhile, it was confirmed that this fraction damaged the bacterial cell membrane structure, leading to the leakage of intracellular proteins in bacteria and thus impairing their infectivity. Network pharmacology analysis and molecular docking results indicated that B. hemsleyana bark’s biologically active compounds (Calenduloside E, Limonin, Acanthoside B, Dihydroberberine) antibacterial activity by regulating cytokines and cell apoptosis, thereby coordinating the body’s microbial homeostasis and inflammation. Additionally, BNB significantly reduced the secretion of the inflammatory cytokines IL-1β, TNF-α and IL-6 in vitro via the NF-κB pathway. The crude extract from the bark of B. hemsleyana has antibacterial and Anti-inflammatory activity. The n-butanol fraction showed a significant antibacterial effect on P. gingivalis. Full article
(This article belongs to the Section Phytochemistry)
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18 pages, 7199 KB  
Article
Zein/Shellac Composite Films with Limonin and Resveratrol: Characterization and Application for Strawberry Preservation
by Limin Wang, Qianfei Jia, Yuxi Qin, Shufen Li and Zijian Wu
Foods 2026, 15(1), 83; https://doi.org/10.3390/foods15010083 - 26 Dec 2025
Viewed by 428
Abstract
This study aimed to fabricate an active film incorporating limonin (LM) and resveratrol (RES) within a zein/shellac (ZS) matrix for use in strawberry preservation. Zein/shellac composite films embedded with varying concentrations of LM (0–4% w/w) and RES were successfully fabricated [...] Read more.
This study aimed to fabricate an active film incorporating limonin (LM) and resveratrol (RES) within a zein/shellac (ZS) matrix for use in strawberry preservation. Zein/shellac composite films embedded with varying concentrations of LM (0–4% w/w) and RES were successfully fabricated using coaxial electrospinning. The prepared films were comprehensively characterized for their mechanical properties, water vapor permeability (WVP), antioxidant capacity (DPPH, ABTS, FRAP), and antibacterial efficacy against E. coli and S. aureus. Mechanical properties and WVP results revealed that the ZSLM4R film exhibited an elongation at break (EBA) of 28.91%, tensile strength (TS) of 0.93 MPa, elastic modulus (EM) of 40.76 MPa, and a WVP of 1.55 g mm/m2. h. kPa. Furthermore, LM and RES increased the antioxidant properties of the composite film. ZSLM4R’s free radical scavenging activities against DPPH and ABTS were 68.14% and 89.69%, respectively. The composite film also demonstrated strong antibacterial efficacy against E. coli and S. aureus. When applied to strawberries, ZSLM4R packaging effectively extended the fruit’s shelf life compared to the unwrapped and commercial polyethylene (PE) controls. These obtained results indicate that LM/RES-incorporated zein/shellac composites are a promising eco-friendly packaging alternative for preserving perishable fresh produce and extending its shelf life. Full article
(This article belongs to the Section Food Packaging and Preservation)
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21 pages, 1870 KB  
Article
Effects of Different Citrus Varieties and Harvesting Time on the Quality of Citrus Dark Tea
by Fuwei Guo, Yuanfang Jiao, De Zhang, Zhi Yu, Dejiang Ni, Han Huang and Yuqiong Chen
Foods 2025, 14(18), 3181; https://doi.org/10.3390/foods14183181 - 12 Sep 2025
Viewed by 1962
Abstract
The genus Citrus consists of Tangerines (Citrus × reticulata), Mandarins (Citrus × reticulata), Oranges (Citrus × sinensis), Grapefruits (Citrus maxima), Lemons (Citrus × limon), and other citrus fruits. This study investigated the effects [...] Read more.
The genus Citrus consists of Tangerines (Citrus × reticulata), Mandarins (Citrus × reticulata), Oranges (Citrus × sinensis), Grapefruits (Citrus maxima), Lemons (Citrus × limon), and other citrus fruits. This study investigated the effects of 74 different citrus species and harvesting time on the quality of citrus dark tea using Hubei green brick tea as the raw material. Oranges (Citrus × sinensis) were found to outperform other species in improving the quality of citrus dark tea. Additionally, the impact of orange harvesting time (August to December) on the quality of orange dark tea was investigated by using the Peach leaf orange with the highest sensory score as the subject of the study. Results revealed that harvesting time can considerably impact the flavor quality of orange dark tea, but not its infusion color or scent. Specifically, the orange dark tea processed with September-harvested Peach leaf orange exhibited a strong fruity scent, a sweet and smooth flavor, and a harmonious tea and fruit aroma. With the delay of harvesting time, a downtrend was observed in the content of soluble proteins, flavonoids, hesperidin, synephrine, and limonin, as well as total volatile components, with the largest decrease in synephrine and flavonoids. However, the polysaccharide content increased in the peel of Peach leaf orange. Functional analysis revealed that the delay of harvesting time can diminish the inhibitory capacity of orange-dark tea on α-glucosidase and α-amylase. Organoleptic quality and physiological activity analysis demonstrated September as the optimal time for harvesting Peach leaf oranges for processing orange dark tea. Full article
(This article belongs to the Section Food Quality and Safety)
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17 pages, 5549 KB  
Article
The Effects of Limonin, Myo-Inositol, and L-Proline on the Cryopreservation of Debao Boar Semen
by Qianhui Feng, Yanyan Yang, Bing Zhang, Wen Shi, Yizhen Fang, Chunrong Xu, Zhuxin Deng, Wanyou Feng and Deshun Shi
Animals 2025, 15(15), 2204; https://doi.org/10.3390/ani15152204 - 27 Jul 2025
Cited by 2 | Viewed by 1142
Abstract
Semen cryopreservation is associated with sperm vulnerability to oxidative stress and ice crystal-induced damage, adversely affecting in vitro fertilization (IVF) success. This study aimed to investigate the effects of freezing diluent supplemented with antioxidant limonin (Lim), myo-inositol (MYO), and the ice crystal formation [...] Read more.
Semen cryopreservation is associated with sperm vulnerability to oxidative stress and ice crystal-induced damage, adversely affecting in vitro fertilization (IVF) success. This study aimed to investigate the effects of freezing diluent supplemented with antioxidant limonin (Lim), myo-inositol (MYO), and the ice crystal formation inhibitor L-proline (LP) through sperm motility, morphological integrity, and antioxidant capacity. The Lim (150 mM), MYO (90 mM), and LP (100 mM) significantly ameliorated the quality of post-thaw sperm in Debao boar, and combined treatment of these agents significantly enhanced sperm motility, structural integrity, and antioxidant capacity compared with individual agents (p < 0.05). Notably, the combined use of these agents reduced glycerol concentration in the freezing diluent from 3% to 2%. Meanwhile, the integrity of the sperm plasma membrane, acrosome membrane, and mitochondrial membrane potential was significantly improved (p < 0.05), and the result of IVF revealed the total cell count of the blastocysts was also greater in the 2% glycerol group (p < 0.05). In conclusion, the newly developed freezing diluent for semen, by adding Lim (150 mM), MYO (90 mM), and LP (100 mM), can enhance the quality of frozen–thawed Debao boar sperm and reduce the concentration of glycerol from 3% to 2% as high concentrations of glycerol can impair the quality of thawed sperm and affect in vitro fertilization outcomes. In conclusion, the improved dilution solution formulated demonstrated efficacy in enhancing the quality of porcine spermatozoa following cryopreservation and subsequent thawing. Full article
(This article belongs to the Section Pigs)
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16 pages, 3392 KB  
Article
Inhibitory Effects of Phellodendri Cortex Against Airway Inflammation and Hyperresponsiveness in Ovalbumin-Induced Murine Asthma Model
by Seong-Kyeom Kim, Ji-Won Do, Seong-Kyun Lee, Jae-Ho Park, Ju-Hyoung Kim and Heung-Bin Lim
Molecules 2025, 30(8), 1795; https://doi.org/10.3390/molecules30081795 - 16 Apr 2025
Cited by 3 | Viewed by 3436
Abstract
Phellodendri cortex (PC), the dried trunk bark of Phellodendron amurense RUPR, has traditionally been used to treat patients who suffer from gastroenteritis, abdominal pain or diarrhea. Its major bioactive compounds include alkaloids and limonin, and many physiological activities including anti-microbial, anti-ulcer and anti-cancer [...] Read more.
Phellodendri cortex (PC), the dried trunk bark of Phellodendron amurense RUPR, has traditionally been used to treat patients who suffer from gastroenteritis, abdominal pain or diarrhea. Its major bioactive compounds include alkaloids and limonin, and many physiological activities including anti-microbial, anti-ulcer and anti-cancer as well as anti-inflammation have been reported. Although PC is an effective anti-inflammatory natural substance that inhibits the inflammatory response, its effect on allergic asthma has not yet been investigated. The aim of this study was to evaluate the anti-asthmatic effects of PC in an ovalbumin (OVA)-induced murine model of asthma. As a result, PC inhibited airway eosinophil accumulation, the influx of inflammatory cells, airway hyperresponsiveness (AHR), production of Th2 cytokines (IL-4, IL-5 and IL-13) and tumor necrosis factor-α (TNF-α) in the bronchoalveolar lavage fluid and/or lung, as well as OVA-specific immunoglobulin E (IgE) in the serum. Furthermore, PC suppressed the gene expression of IL-4, IL-5, IL-13, TARC and CCR3, and attenuated unique histological changes that are associated with airway inflammatory reactions including the infiltration of various inflammatory cells, collagen deposition and goblet cell hyperplasia in lung tissues. These results indicate that PC may have preventive and/or therapeutic effects for allergic asthma via the inhibition of cytokines, chemokines and chemokine receptors associated with allergic inflammation. Full article
(This article belongs to the Section Natural Products Chemistry)
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20 pages, 3380 KB  
Article
Bioactive Compounds and Bitterness Properties of Newly Developed Interspecific Citrus Hybrids (Citrus maxima [Burm. f.] Osbeck × Citrus sinensis [L.] Osbeck)
by Narendra Singh, Radha Mohan Sharma, Anil Kumar Dubey, Supradip Saha, Om Prakash Awasthi, Chellapilla Bharadwaj, Amitha Mithra Sevanthi, Amrender Kumar, Nimisha Sharma, Rakesh Kumar, Vijay Kumar, Anil Kumar Yadav, Mukesh Shivran, Ron Porat and Deepak
Horticulturae 2025, 11(2), 208; https://doi.org/10.3390/horticulturae11020208 - 15 Feb 2025
Cited by 4 | Viewed by 2579
Abstract
In the present study, variation in phytochemical properties was estimated in 16 newly developed interspecific citrus hybrids (Citrus maxima [Burm. f.] Osbeck × C. sinensis [L.] Osbeck) and parental genotypes. Results showed that the interspecific hybridization of pummelo with sweet orange significantly [...] Read more.
In the present study, variation in phytochemical properties was estimated in 16 newly developed interspecific citrus hybrids (Citrus maxima [Burm. f.] Osbeck × C. sinensis [L.] Osbeck) and parental genotypes. Results showed that the interspecific hybridization of pummelo with sweet orange significantly improved the fruit quality traits. Ascorbic acid, total phenol, total flavonoids, total carotenoid, and lycopene content in the fruit juice of studied genotypes varied from 37.8 to 72.02 mg/100 mL, 19.63–112.59 GAE mg/100 mL, 1.09–2.39 QE mg/100 mL, 34.6–519.81 μg/100 mL, and 17.59–395.71 μg/100 mL, respectively. Among the new hybrids, the highest antioxidant value (DPPH assay) was recorded as 2.53 TE μmol/mL as against the 2.22 TE μmol/mL in sweet orange cv. Mosambi. The citrus hybrid genotypes SCSH-9-2/12, SCSH-9-10/12, SCSH-11-9/13, SCSH-11-15/12, and SCSH-17-19/13 proved superior for studied phytochemicals. Pearson’s correlation and principal component analyses revealed the association among the traits and key components underlying the genetic variations. The bitterness properties were studied by quantifying naringin and limonin content in the fresh and stored juice (24 h storage at 4 °C) in addition to the sensory evaluation. The new hybrid SCSH-9-2/12, SCSH-9-10/12, and SCSH-11-9/13 have very low limonin content (<1.0 mg/L) with the least delayed bitterness properties, thus proving their potential for juice processing and storage. Full article
(This article belongs to the Special Issue Fruits Quality and Sensory Analysis—2nd Edition)
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18 pages, 7089 KB  
Article
Limonin Exhibits Anti-Inflammatory Effects by Inhibiting mTORC1 and Mitochondrial Reactive Oxygen Species in Psoriatic-like Skin Inflammation
by Seung Taek Lee, Jong Yeong Lee, Ha Eun Kim, Jun-Young Park and Jin Kyeong Choi
Antioxidants 2024, 13(12), 1541; https://doi.org/10.3390/antiox13121541 - 16 Dec 2024
Cited by 2 | Viewed by 2749
Abstract
Psoriasis is a chronic inflammatory skin disorder characterized by abnormal immune responses and keratinocyte hyperproliferation. Limonin, a bioactive compound found in citrus fruits, has anti-inflammatory properties in various models; however, its effects on psoriasis are not fully understood. We investigated the therapeutic potential [...] Read more.
Psoriasis is a chronic inflammatory skin disorder characterized by abnormal immune responses and keratinocyte hyperproliferation. Limonin, a bioactive compound found in citrus fruits, has anti-inflammatory properties in various models; however, its effects on psoriasis are not fully understood. We investigated the therapeutic potential of limonin in a 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced psoriasis mouse model. Mice were treated with TPA to induce psoriasis-like skin lesions, followed by intraperitoneal administration of limonin (200 or 400 μg/mouse) for six days. The results showed that limonin improved psoriasis-related symptoms in a psoriasis-like mouse model by suppressing the mRNA expression of pro-inflammatory cytokines and inflammation-related antimicrobial peptides and regulating the expansion of myeloid cells and T cells. Specifically, limonin reduced glucose uptake and oxidative phosphorylation to shift the metabolic program in the inflamed skin cells of psoriasis-like mice. Limonin activates AMPK and proteins related to mTOR inhibition, thereby suppressing the mTOR signaling pathway. It also inhibits mitochondrial mass and mitochondrial ROS production, thereby preventing the development of dysfunctional mitochondria in inflamed skin cells. Overall, limonin modulates key immune responses and metabolic pathways related to inflammation and mitochondrial health in psoriasis. Therefore, it is a promising natural candidate for the treatment of psoriasis and various inflammatory skin diseases. Full article
(This article belongs to the Special Issue Antioxidants for Skin Health)
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19 pages, 2563 KB  
Article
Effectively Enhancing the Physiological Activity and Sensory Quality of Whole Calamondin Puree via Yeast Fermentation
by Hongjian Zhang, Shuaiguang Liu, Zewei Ma, Huan Huang, Lianhe Zheng, Yan Tian and Qiuping Zhong
Int. J. Mol. Sci. 2024, 25(22), 11984; https://doi.org/10.3390/ijms252211984 - 7 Nov 2024
Cited by 5 | Viewed by 2540
Abstract
To explore the feasibility of pure yeast fermentation in whole Calamondin puree (FWCP) for the utilization of the whole fruit, yeasts were isolated from naturally fermented Calamondin, and their fermentation characteristics were evaluated. The results indicated that all yeasts were able to ferment [...] Read more.
To explore the feasibility of pure yeast fermentation in whole Calamondin puree (FWCP) for the utilization of the whole fruit, yeasts were isolated from naturally fermented Calamondin, and their fermentation characteristics were evaluated. The results indicated that all yeasts were able to ferment FWCP, reducing the sour taste by degrading citric acid, increasing the contents of nutrients (such as phenols and limonins) and volatile compounds, and enhancing the antioxidant activity and inhibition of α-glucosidase activity (p < 0.05). Among them, P. terricola QJJY1 and H. opuntiae QJJY14 exhibited stronger abilities to degrade organic acids, with P. terricola QJJY1 enhancing the antioxidant capacity by releasing phenolic compounds such as phloretin-3′,5′-di-c-β-glucoside, vitexin-2″-O-rhamnoside, and isomargaritene. Moreover, H. opuntiae QJJY15 improved the contents of characteristic volatile compounds such as terpene hydrocarbons and higher alcohols. In total, 70 components were identified as differential metabolites based on their fold change in the metabolites, with 42 differential metabolites involved in 29 metabolic pathways across four strains. The main pathways related to phenol and flavor enrichment were flavonoid, flavone, and flavonol biosynthesis, monoterpenoid biosynthesis, and glyoxylate and dicarboxylate metabolism. Therefore, yeast fermentation is an effective method for utilizing whole Calamondin. Full article
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17 pages, 6991 KB  
Article
Metabolomic Profiling Reveals the Quality Variations in Citri Reticulatae Pericarpium (Citrus reticulata Blanco cv. Chachiensis) with Different Storage Ages in Response to “Candidatus Liberibacter Asiaticus” Infection
by Jiayin Liang, Yuqing Xi, Jiaming Li, Shugui Xu, Yongqin Zheng, Meirong Xu, Zheng Zheng and Xiaoling Deng
Foods 2024, 13(6), 827; https://doi.org/10.3390/foods13060827 - 8 Mar 2024
Cited by 6 | Viewed by 3139
Abstract
Citri Reticulatae Pericarpium, especially the pericarp of Citrus reticulata Blanco cv. Chachiensis (PCRC), is an important edible and medicinal ingredient for health and pharmacological properties. Citrus Huanglongbing, a devastating disease that currently threatens the citrus industry worldwide, is caused by a phloem-limited alpha-proteobacterium, [...] Read more.
Citri Reticulatae Pericarpium, especially the pericarp of Citrus reticulata Blanco cv. Chachiensis (PCRC), is an important edible and medicinal ingredient for health and pharmacological properties. Citrus Huanglongbing, a devastating disease that currently threatens the citrus industry worldwide, is caused by a phloem-limited alpha-proteobacterium, “Candidatus Liberibacter asiaticus” (CLas). The industry of cultivar Chachiensis has been suffering from HLB. Although HLB affected the quality of citrus fruit, whether the quality of PCRC was affected by HLB remains unclear. In this study, we compared the metabolite profiles between HLB-affected and healthy PCRC from three sources: fresh, 6-month-old, and 9-year-old PCRC, through the untargeted LC–MS method. Compared to healthy controls, various types of bioactive compounds, mainly flavonoids, terpenoids, alkaloids, coumarins, polysaccharides, and phenolic acids, accumulated in HLB-affected PCRC, especially in the HLB-affected 9-year PCRC. In particular, isorhamnetin, isoliquiritigenin, luteolin 7-O-beta-D-glucoside, limonin, geniposide, pyrimidodiazepine, scoparone, chitobiose, m-coumaric acid, malonate, and pantothenic acid, which contributed to the pharmacological activity and health care effects of PCRC, were highly accumulated in HLB-affected 9-year-old PCRC compared to the healthy control. Multibioassay analyses revealed that HLB-affected 9-year-old PCRC had a higher content of total flavonoids and total polyphenols and exhibited similar antioxidant capacity as compared to healthy controls. The results of this study provided detailed information on the quality of HLB-affected PCRC. Full article
(This article belongs to the Section Nutraceuticals, Functional Foods, and Novel Foods)
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11 pages, 1569 KB  
Article
Oral Ingestion of Yuzu Seed Oil Suppresses the Development of Atopic Dermatitis-like Skin Lesions in NC/Nga Mice
by Kimito Asano, Yoshiya Watanabe, Mio Miyamoto, Mochifumi Toutani and Shunji Mizobuchi
Int. J. Mol. Sci. 2024, 25(5), 2689; https://doi.org/10.3390/ijms25052689 - 26 Feb 2024
Cited by 1 | Viewed by 2767
Abstract
Long-term oral ingestion of unheated yuzu seed oil in humans reduces lipid peroxides in the blood. Moreover, yuzu seed oil contains limonin, which can induce antioxidant and anti-inflammatory effects by activating the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2). Previously, Nrf2 [...] Read more.
Long-term oral ingestion of unheated yuzu seed oil in humans reduces lipid peroxides in the blood. Moreover, yuzu seed oil contains limonin, which can induce antioxidant and anti-inflammatory effects by activating the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2). Previously, Nrf2 has been shown to reduce atopic dermatitis (AD). Therefore, we hypothesized that ingesting unheated yuzu seed oil can regulate AD through Nrf2. An AD model was established using NC/Nga mice through repeated local exposure to mite antigens. Unheated and purified yuzu seed oil (100 µL/mice) or water (control, 100 µL/mice) was administered orally once a day using a gastric cannula for rodents for 28 days. On day 28, mice in the unheated yuzu seed oil group exhibited significantly lower clinical skin severity scores and ear thickness than those in the purified yuzu seed oil and water groups. Serum histamine levels remained unaltered among the three AD-induced groups. Serum Dermatophagoides farina body (Dfb)-specific immunoglobulin E (IgE) levels were significantly lower in the unheated yuzu seed oil group. Oral ingestion of yuzu seed oil in NC/Nga AD model mice significantly suppressed dermatitis deterioration and decreased serum IgE levels. Clinical trials (n = 41) have already confirmed that unheated yuzu oil is safe for long-term intake, further suggesting its potential use in improving AD symptoms. Full article
(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)
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17 pages, 4236 KB  
Article
Selection of Reference Genes in Evodia rutaecarpa var. officinalis and Expression Patterns of Genes Involved in Its Limonin Biosynthesis
by Yu Zhou, Yuxiang Zhang, Detian Mu, Ying Lu, Wenqiang Chen, Yao Zhang, Ruiying Zhang, Ya Qin, Jianhua Yuan, Limei Pan and Qi Tang
Plants 2023, 12(18), 3197; https://doi.org/10.3390/plants12183197 - 7 Sep 2023
Cited by 10 | Viewed by 2743
Abstract
E. rutaecarpa var. officinalis is a traditional Chinese medicinal plant known for its therapeutic effects, which encompass the promotion of digestion, the dispelling of cold, the alleviation of pain, and the exhibition of anti-inflammatory and antibacterial properties. The principal active component of this [...] Read more.
E. rutaecarpa var. officinalis is a traditional Chinese medicinal plant known for its therapeutic effects, which encompass the promotion of digestion, the dispelling of cold, the alleviation of pain, and the exhibition of anti-inflammatory and antibacterial properties. The principal active component of this plant, limonin, is a potent triterpene compound with notable pharmacological activities. Despite its significance, the complete biosynthesis pathway of limonin in E. rutaecarpa var. officinalis remains incompletely understood, and the underlying molecular mechanisms remain unexplored. The main purpose of this study was to screen the reference genes suitable for expression analysis in E. rutaecarpa var. officinalis, calculate the expression patterns of the genes in the limonin biosynthesis pathway, and identify the relevant enzyme genes related to limonin biosynthesis. The reference genes play a pivotal role in establishing reliable reference standards for normalizing the gene expression data, thereby ensuring precision and credibility in the biological research outcomes. In order to identify the optimal reference genes and gene expression patterns across the diverse tissues (e.g., roots, stems, leaves, and flower buds) and developmental stages (i.e., 17 July, 24 August, 1 September, and 24 October) of E. rutaecarpa var. officinalis, LC-MS was used to analyze the limonin contents in distinct tissue samples and developmental stages, and qRT-PCR technology was employed to investigate the expression patterns of the ten reference genes and eighteen genes involved in limonin biosynthesis. Utilizing a comprehensive analysis that integrated three software tools (GeNorm ver. 3.5, NormFinder ver. 0.953 and BestKeeper ver. 1.0) and Delta Ct method alongside the RefFinder website, the best reference genes were selected. Through the research, we determined that Act1 and UBQ served as the preferred reference genes for normalizing gene expression during various fruit developmental stages, while Act1 and His3 were optimal for different tissues. Using Act1 and UBQ as the reference genes, and based on the different fruit developmental stages, qRT-PCR analysis was performed on the pathway genes selected from the “full-length transcriptome + expression profile + metabolome” data in the limonin biosynthesis pathway of E. rutaecarpa var. officinalis. The findings indicated that there were consistent expression patterns of HMGCR, SQE, and CYP450 with fluctuations in the limonin contents, suggesting their potential involvement in the limonin biosynthesis of E. rutaecarpa var. officinalis. This study lays the foundation for further research on the metabolic pathway of limonin in E. rutaecarpa var. officinalis and provides reliable reference genes for other researchers to use for conducting expression analyses. Full article
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82 pages, 36773 KB  
Systematic Review
An Insight into Citrus medica Linn.: A Systematic Review on Phytochemical Profile and Biological Activities
by Nadia Benedetto, Vittorio Carlucci, Immacolata Faraone, Ludovica Lela, Maria Ponticelli, Daniela Russo, Claudia Mangieri, Nikolay T. Tzvetkov and Luigi Milella
Plants 2023, 12(12), 2267; https://doi.org/10.3390/plants12122267 - 10 Jun 2023
Cited by 32 | Viewed by 8206
Abstract
Plant species are a reservoir of natural compounds that can potentially be used to treat different diseases. Citrus medica Linn. belonging to the Rutaceae family, has been used for centuries in medicine for its antioxidant, anti-inflammatory, antimicrobial, antiviral, and antihyperglycemic properties. These activities [...] Read more.
Plant species are a reservoir of natural compounds that can potentially be used to treat different diseases. Citrus medica Linn. belonging to the Rutaceae family, has been used for centuries in medicine for its antioxidant, anti-inflammatory, antimicrobial, antiviral, and antihyperglycemic properties. These activities are ascribable not only to the presence of health-promoting macronutrients and micronutrients, such as carbohydrates, minerals, amino acids, and vitamins, but also to specialized metabolites, such as flavonoids (apigenin, hesperetin, hesperidin, naringin, naringenin, rutin, quercetin, and diosmin), coumarins (citropten, scoparone, and bergapten), terpenes (limonene, γ-terpinene, limonin, and nomilin), and phenolic acids (p-coumaric acid, trans-ferulic acid, and chlorogenic acid). In recent years, particular attention has been focused on the antioxidant, anti-inflammatory, antimicrobial activity, antidiabetic, anticancer, and neuroprotective activity of C. medica. However, although many studies have reported this species’ chemical and biological properties, the literature has never been analyzed via a systematic approach. For this reason, using PubMed and Scopus as databases, we performed a systematic review of C. medica’s chemical composition and biological properties to inspire new research approaches and increase its curative application. Full article
(This article belongs to the Special Issue Multifunctionality of Phenolic Compounds in Plants)
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18 pages, 4918 KB  
Article
Molecular Docking and Simulation-Binding Analysis of Plant Phytochemicals with the Hepatocellular Carcinoma Targets Epidermal Growth Factor Receptor and Caspase-9
by Ghulam Mustafa, Shumaila Younas, Hafiza Salaha Mahrosh, Mohammed Fahad Albeshr and Eijaz Ahmed Bhat
Molecules 2023, 28(8), 3583; https://doi.org/10.3390/molecules28083583 - 20 Apr 2023
Cited by 27 | Viewed by 8353
Abstract
Among primary liver cancers, hepatocellular carcinoma (HCC) is one of the most common forms and it has been categorized as the joint-fourth largest reason of cancer-related deaths globally. Different factors such as alcohol abuse, hepatitis B and C, viral infections, and fatty liver [...] Read more.
Among primary liver cancers, hepatocellular carcinoma (HCC) is one of the most common forms and it has been categorized as the joint-fourth largest reason of cancer-related deaths globally. Different factors such as alcohol abuse, hepatitis B and C, viral infections, and fatty liver diseases are mainly related to the pathogenesis of HCC. In the current study, 1000 total various plant phytochemicals were docked to proteins involved in HCC. The compounds were docked to the active site amino acids of epidermal growth factor receptor and caspase-9 as receptor proteins in order to explore their inhibiting potential. The top five compounds against each receptor protein were explored as potential drug candidates on the basis of their binding affinity and root-mean square deviation values. The top two compounds against each protein were found to be liquoric acid (S-score −9.8 kcal/mol) and madecassic acid (S-score −9.3 kcal/mol) against EGFR, and limonin (S-score −10.5 kcal/mol) and obamegine (S-score −9.3 kcal/mol) against the caspase-9 protein. The selected phytochemicals were further assessed through drug scanning using Lipinski’s rule of five to explore their molecular properties and druggability. According to the ADMET analysis, the selected phytochemicals were found to be non-toxic and non-carcinogenic. Finally, the molecular dynamics simulation study revealed that liquoric acid and limonin were stabilized within the binding pockets of EGFR and capase-9, respectively, and stayed firmly bound throughout the simulation. In light of the current findings, the phytochemicals reported in this study, especially liquoric acid and limonin, could be used as potential drugs for the treatment of HCC in the future. Full article
(This article belongs to the Special Issue Natural Compounds in Modern Therapies)
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20 pages, 10439 KB  
Article
Metabolomic Analysis of Phytochemical Compounds from Ethanolic Extract of Lime (Citrus aurantifolia) Peel and Its Anti-Cancer Effects against Human Hepatocellular Carcinoma Cells
by Pakkapong Phucharoenrak, Chawanphat Muangnoi and Dunyaporn Trachootham
Molecules 2023, 28(7), 2965; https://doi.org/10.3390/molecules28072965 - 26 Mar 2023
Cited by 21 | Viewed by 7333
Abstract
Lime peels are food waste from lime product manufacturing. We previously developed and optimized a green extraction method for hesperidin-limonin-rich lime peel extract. This study aimed to identify the metabolomics profile of phytochemicals and the anti-cancer effects of ethanolic extract of lime ( [...] Read more.
Lime peels are food waste from lime product manufacturing. We previously developed and optimized a green extraction method for hesperidin-limonin-rich lime peel extract. This study aimed to identify the metabolomics profile of phytochemicals and the anti-cancer effects of ethanolic extract of lime (Citrus aurantifolia) peel against liver cancer cells PLC/PRF/5. The extract’s metabolomics profile was analyzed by using LC-qTOF/MS and GC-HRMS. The anti-cancer effects were studied by using MTT assay, Annexin-PI assay, and Transwell-invasion assay. Results show that the average IC50(s) of hesperidin, limonin, and the extract on cancer cells’ viability were 165.615, 188.073, and 503.004 µg/mL, respectively. At the IC50 levels, the extract induced more apoptosis than those of pure compounds when incubating for 24 and 48 h (p < 0.0001). A combination of limonin and hesperidin showed a synergistic effect on apoptosis induction (p < 0.001), but the effect of the combination was still less than that of the extract at 48 h. Furthermore, the extract significantly inhibited cancer cell invasion better than limonin but equal to hesperidin. At the IC50 level, the extract contains many folds lower amounts of hesperidin and limonin than the IC50 doses of the pure compounds. Besides limonin and hesperidin, there were another 60 and 22 compounds detected from the LCMS and GCMS analyses, respectively. Taken altogether, the superior effect of the ethanolic extract against liver cancer cells compared to pure compound likely results from the combinatorial effects of limonin, hesperidin, and other phytochemical components in the extract. Full article
(This article belongs to the Special Issue Biological Activity of Plant Compounds and Extracts, 2nd Edition)
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21 pages, 2310 KB  
Article
The New Insight into the Effects of Different Fixing Technology on Flavor and Bioactivities of Orange Dark Tea
by Yuanfang Jiao, Yulin Song, Zhi Yan, Zhuanrong Wu, Zhi Yu, De Zhang, Dejiang Ni and Yuqiong Chen
Molecules 2023, 28(3), 1079; https://doi.org/10.3390/molecules28031079 - 20 Jan 2023
Cited by 11 | Viewed by 4058
Abstract
Peach leaf orange dark tea (ODT) is a fruity tea made by removing the pulp from peach leaf orange and placing dry Qingzhuan tea into the husk, followed by fixing them together and drying. Since the quality of traditional outdoor sunlight fixing (SL) [...] Read more.
Peach leaf orange dark tea (ODT) is a fruity tea made by removing the pulp from peach leaf orange and placing dry Qingzhuan tea into the husk, followed by fixing them together and drying. Since the quality of traditional outdoor sunlight fixing (SL) is affected by weather instability, this study explored the feasibility of two new fixing methods, including hot air fixing (HA) and steam fixing (ST). Results showed that fixing method had a great impact on ODT shape, aroma, and taste. Compared with SL and ST, HA endowed ODT with higher fruit aroma, mellow taste, better coordination, and higher sensory evaluation score. Physical–chemical composition analysis showed that SL-fixed orange peel was higher than HA- or ST-fixed peel in the content of polyphenols, flavonoids, soluble protein, hesperidin and limonin, while HA has a higher content of volatile substances and contains more alcohols, aldehydes and ketones, and acid and esters than ST and SL. Activity analysis showed that HA was superior to ST or SL in comprehensive antioxidant activity and inhibitory activity against α-glucosidase. Comprehensive results demonstrated that HA has better performance in improving ODT quality and can replace the traditional SL method in production. Full article
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