Polymethoxylated flavone variations and in vitro biological activities of locally cultivated Citrus varieties in China.

Citrus peels are rich in polymethoxylated flavones (PMFs), which have beneficial health and pharmacological properties. In this study, the profiles, variations, and biological activities of PMFs in the peel extracts of 27 Citrus varieties (eight species) native to China were investigated. UPLC-QTOF-MS/MS analysis revealed that mandarin accumulated more diversity and higher detectable PMF contents. Wangcangzhoupigan (ZPG) possessed the highest antioxidant capacity. Gailiangcheng (GLC) and Bingtangcheng (BTC), sweet oranges showed excellent inhibitory effects against pancreatic lipase and α-glucosidase, respectively. Most citrus extracts effectively inhibited the production of ROS and pro-inflammatory cytokines, while increasing the accumulation of anti-inflammatory cytokines. In addition, Limeng (LM), Cupig-oushigan (GSG), and Yanxiwanlu (YXWL) showed anti-proliferative effects against DU145 and PC3 cancer cells. This study provides a comprehensive PMF profile and biological activities of various citrus species and will benefit future functional citrus breeding practices aimed at designing plants rich in total or specific PMFs for health benefits.

Nobiletin enhances the antifungal activity of eugenol nanoemulsion against Penicillium italicum in both in vitro and in vivo settings.

The study prepared and used eugenol nanoemulsion loaded with nobiletin as fungistat to study its antifungal activity and potential mechanism of Penicillium italicum (P. italicum). The results showed that the minimum inhibitory concentration (MIC) of eugenol nanoemulsion loaded with nobiletin (EGN) was lower than that of pure eugenol nanoemulsion (EG), which were 160 µg/mL and 320 µg/mL, respectively. At the same time, the mycelial growth inhibition rate of EGN nanoemulsion (54.68 %) was also higher than that of EG nanoemulsion (9.92 %). This indicates that EGN nanoemulsion is more effective than EG nanoemulsion. Compared with EG nanoemulsion, the treatment of EGN nanoemulsion caused more serious damage to the cell structure of P. italicum. At the same time, in vitro inoculation experiments found that EGN nanoemulsion has better control and delay the growth and reproduction of P. italicum in citrus fruits. And the results reflected that EGN nanoemulsion may be considered as potential resouces of natural antiseptic to inhibit blue mold disease of citrus fruits, because it has good antifungal activity.

Effect of polyethoxylated flavonoids (PMFs)-loaded citral and chitosan composite coatings on citrus preservation: From the perspective of fruit resistance.

Previous studies have shown that polymethoxylated flavonoids-loaded citral emulsion (PCT) can inhibit the growth and reproduction of Penicillium in citrus; however, PCT is difficult to apply to fruit preservation due to its high fluidity and volatility. Therefore, in this study, we combined PCT with chitosan (CS) to investigate the effect of a composite coating on citrus preservation. The results showed that compared to the control group, the CS-PCT group could effectively reduce the decay rate and maintain moisture availability, color difference, and hardness. Moreover, the contents of nonenzymatic antioxidants and volatile substances with antimicrobial activity were better preserved. In addition, the activities of related antioxidant enzymes were greater in the treatment group, and the expression of the corresponding enzyme-encoding genes was upregulated. Consequently, CS-PCT treatment could effectively maintain fruit quality and improve the resistance of citrus fruits during storage; moreover, it can be considered a nontoxic and efficient citrus preservative.

A large presence/absence variation in the promotor of the ClLOG gene determines trichome elongation in watermelon.

KEY MESSAGE: The ClLOG gene encoding a cytokinin riboside 5'-monophosphate phosphoribohydrolase determines trichome length in watermelon, which is associated with its promoter variations. Trichomes, which are differentiated from epidermal cells, are special accessory structures that cover the above-ground organs of plants and possibly contribute to biotic and abiotic stress resistance. Here, a bulked segregant analysis (BSA) of an F2 population with significant variations in trichome length was undertaken. A 1.84-Mb candidate region on chromosome 10 was associated with trichome length. Resequencing and fine-mapping analyses indicated that a 12-kb structural variation in the promoter of Cla97C10G203450 (ClLOG) led to a significant expression difference in this gene in watermelon lines with different trichome lengths. In addition, a virus-induced gene silencing analysis confirmed that ClLOG positively regulated trichome elongation. These findings provide new information and identify a potential target gene for controlling multicellular trichome elongation in watermelon.

Citrus Carotenoid Extracts Promote ROS Accumulation and Induce Oxidative Stress to Exert Anti-Proliferative and Pro-Apoptotic Effects in MDA-MB-231 Cells.

Citrus fruits are economically significant crops worldwide, as they contain various bioactive compounds that possess health-promoting properties. Carotenoids, as the most important component in citrus, exhibit notable pharmacological activities, such as antioxidation and anticancer, which make carotenoids valuable in the prevention and treatment of breast cancer. In this study, after treatment with carotenoid extracts from XiYou (XY) and ZaoHongQiCheng (ZH), we evaluated the cytotoxicity, apoptosis, antioxidant system, and oxidative stress induced by ROS overproduction and MMP damage in MDA-MB-231 cells. The analysis confirmed that cell proliferation was inhibited in a concentration-dependent manner, accompanied by G0/G1 arrest and cell apoptosis. XY and ZH promoted the accumulation of ROS, decreased MMP, increased malondialdehyde (MDA) levels, consumed glutathione (GSH), and reduced the activity of antioxidant enzymes (peroxidase (POD), catalase (CAT), glutathione reductase (GR), and superoxide dismutase (SOD)). Meanwhile, XY and ZH induced apoptosis through the mitochondrial pathway by significantly upregulated P53, BAX, caspase-3, caspase-7, and caspase-9 gene expression levels and downregulated Bcl-2. Carotenoid-rich extracts were found to cause oxidative stress by enhancing ROS production through their pro-oxidative potential, and the aggravation of oxidative processes promotes apoptosis in MDA-MB-231 cells. These results indicate that citrus carotenoids can be used as potential pro-oxidants and have the potential to be developed into products for the prevention or treatment of breast cancer.

Evaluation of Antioxidant Activity and Drug Delivery Potential of Cell-Derived Extracellular Vesicles from Citrus reticulata Blanco cv. 'Dahongpao'.

Plant extracellular vesicles (PEVs) have attracted increasing attention due to their rich composition, good antioxidant and anti-inflammatory activity, and ability to transport drugs. As a common fruit, citrus is an ideal material for extracting PEVs because of the diversity and abundance of bioactive substances in it. In our study, citrus-derived extracellular vesicles (CEVs) were extracted from red mandarin (Citrus reticulata Blanco cv. 'Dahongpao') and it was found that they contain high levels of lipids, proteins, and carbohydrates. The high levels of total phenols and total flavonoids suggest that CEVs have good chemical antioxidant properties. We also demonstrated through cell experiments that CEVs have significant antioxidant and anti-inflammatory effects. Furthermore, we found that CEVs have an encapsulation rate of 71.5 ± 0.19% and a high drug-carrying capacity of 4.96 ± 0.22% and can enhance antioxidant and anti-inflammatory activity when loaded with tangeretin. Our results show that CEVs contain abundant bioactive components, have low toxicity, exhibit good antioxidant and anti-inflammatory properties, and can serve as drug delivery agents. This study has important implications for utilizing citrus materials and developing natural anti-oxidative and anti-inflammatory biomaterials.

Citrus Carotenoid Extracts Exert Anticancer Effects through Anti-Proliferation, Oxidative Stress, and Mitochondrial-Dependent Apoptosis in MCF-7 Cells.

Citrus is a globally popular fruit crop that contains bioactive compounds with numerous health benefits. Carotenoids are one of the main bioactive compounds present in citrus pulp. They possess exceptional antioxidant and anticancer properties, making them potentially effective in the prevention and treatment of breast cancer. Different citrus species, identified as ZMPG, DFGJ, NFMJ, XY, and ZHQC, were studied for their antioxidant activity and anticancer activity. XY had the highest total carotenoid content (75.30 µg/g FW), and ZHQC (ZH) had the lowest carotenoid content (19.74 µg/g FW). The composition of NFMJ, ZMPG, and DFHJ consisted of the most abundant number of carotenoids, while XY only had three types. The antioxidant capacity of the carotenoid extracts was evaluated, and ZH and DFHJ were identified as good sources of antioxidants. XY and ZH significantly inhibited cell proliferation, migration, and arresting cells during the G0/G1 phase. XY and ZH enhanced the accumulation of reactive oxygen species (ROS); reduced mitochondrial membrane potential (MMP); reduced the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and peroxidase (POD); decreased glutathione (GSH) levels; and increased the malonaldehyde (MDA) content. Apoptosis occurred through the mitochondrial-mediated pathway through the up-regulation of BAX, caspase-3, and caspase-9 and the down-regulation of Bcl-2. In this study, the carotenoid-rich extracts of citrus pulp were found to induce oxidative stress through their pro-oxidant potential and regulate cell apoptosis in MCF-7 cancer cells. These results indicate that citrus carotenoids act as pro-oxidants and have the potential to be utilized for the development of anti-breast cancer products.

Comparative Analysis of the Impact of Three Drying Methods on the Properties of Citrus reticulata Blanco cv. Dahongpao Powder and Solid Drinks.

Citrus reticulata Blanco cv. Dahongpao is a traditional Chinese citrus variety. Due to the high investment in storage and transport of Citrus reticulata Blanco cv. Dahongpao and the lack of market demand, the fresh fruit is wasted. The processing of fresh fruit into fruit drinks can solve the problem of storage and transport difficulties and open up new markets. Investigating the effects of different drying processes (hot air, freeze, and spray drying) on fruit powders is a crucial step in identifying a suitable production process. The experiment measured the effects of different drying methods (hot air drying, freeze drying, and spray drying) on the nutrient, bioactive substance, and physical characteristics of fruit powder. This study measured the influence of three different drying methods (hot air, freeze, and spray drying) on the nutritional, bioactive substance, and physical characteristics of fruit powder. The results showed that compared to vacuum freeze-drying at low temperature (-60 °C) and spray-drying at high temperatures (150 °C), hot air drying at 50 °C produced fruit powder with superior nutritional quality, higher levels of active substances, and better physical properties. Hot air drying produced fruit powder that had the highest content of amino acids (11.48 ± 0.08 mg/g DW), vitamin C (112.09 ± 2.86 µg/g DW), total phenols (14.78 ± 0.30 mg/g GAE DW), total flavonoids (6.45 ± 0.11 mg/g RE DW), organic acids, and antioxidant activity capacity. Additionally, this method yielded the highest amounts of zinc (8.88 ± 0.03 mg/Kg DW) and soluble sugars, low water content, high solubility, and brown coloration of the fruit powder and juice. Therefore, hot air drying is one of the best production methods for producing high-quality fruit powder in factory production.

Polymethoxylated flavonoids (PMFs)-loaded citral nanoemulsion controls green mold in citrus by damaging the cell membrane of Penicillium digitatum.

Citrus is susceptible to Penicillium digitatum (P. digitatum) infection in post-harvest storage, resulting in enormous economic losses. This study aimed to investigate the antifungal activity and potential mechanism of the combination of Polymethoxylated flavones (PMFs) and citral (two natural antifungal components derived from citrus) against P. digitatum in vitro and citrus fruit. The results show that PMFs can enhance the antifungal activity of citral nanoemulsion, and PMFs-loaded citral nanoemulsion (PCT) has significant antifungal activity in a concentration-dependent manner. PCT can evidently inhibit spore germination and mycelial growth in vitro, and effectively control the growth of green mold on postharvest citrus fruit. Furthermore, PCT treatment resulted in the alteration of mycelia morphology, accumulation of reactive oxygen species, and membrane lipid peroxidation. These changes can disrupt the normal structure and function of the cell membrane, as evidenced by the reduction of total lipid and ergosterol content in the mycelia and the stronger red fluorescence of the cells emitted after PI staining. Based on the above results, we infer that PCT has a strong inhibitory effect on P. digitatum, and its potential mechanism is related to the destruction of the cell membrane. Therefore, PCT can be considered as a botanical fungicide for citrus preservation.

Comparative effects of five polymethoxyflavones purified from Citrus tangerina on inflammation and cancer.

Although the Citrus tangerina cultivar "Dahongpao" (CTD) has been established as a rich source of polymethoxyflavones (PMFs) with anti-inflammatory and anti-cancer properties, their individual effects on cellular signaling remain to be elucidated. In this study, five major PMFs from the peel of CTD were isolated, including sinensetin, tetramethyl-O-scutellarin (5,6,7,4'-tetramethoxyflavone), nobiletin (5,6,7,8,3', 4'-hexamethoxyflavone), tangeretin (5,6,7,8,4'-pentamethoxyflavone), and 5-demethylnobiletin (5-OH-6,7,8,3',4'-pentamethoxyflavone). These PMFs were found to significantly (p < 0.05) inhibit the production of NO and biomarkers of chronic inflammation (TNF-α and IL-6). Additionally, they effectively suppressed mRNA biomarkers of acute inflammation (Cox-2 and iNOS), and to varying degrees promoted the activation of anti-inflammatory cytokines (IL-4, IL-13, TNF-ß, and IL-10). Among the five PMFs, tangeretin was found to have a considerable anti-proliferative effect on tumor cell lines (PC-3 and DU145) and synergistically enhanced the cytotoxicity of mitoxantrone, partially via activation of the PTEN/AKT pathway. The findings of this study provide valuable insights into the activity of different PMF monomers and advance the understanding of the roles of PMFs in promoting apoptotic and anti-cancer effects.

Antifungal Activity of Polymethoxylated Flavonoids (PMFs)-Loaded Citral Nanoemulsion against Penicillium italicum by Causing Cell Membrane Damage.

A major citrus postharvest pathogen, Penicillium italicum (P. italicum), causes substantial economic losses in citrus. In this study, a citral nanoemulsion containing polymethoxylated flavonoids (PMFs), the antimicrobial compounds from citrus, was prepared. The antifungal activity and potential antifungal mechanisms of the nanoemulsion against P. italicum were evaluated. The results showed that the growth of P. italicum was effectively inhibited by the nanoemulsion, with a minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 62.5 and 250 mg L−1, respectively. The nanoemulsion significantly inhibited spore germination and mycelial growth, and it altered the morphology of P. italicum. In addition, the permeability of the cell membrane increased with increasing nanoemulsion concentrations, as evidenced by a rapid rise in extracellular electric conductivity and stronger red fluorescence from mycelia (propidium iodide staining). Compared with the control, the nanoemulsion treatment induced a decrease in total lipid and ergosterol contents in P. italicum cells by 64.61% and 60.58%, respectively, demonstrating that membrane integrity had been disrupted. The results indicated that the PMFs-loaded nanoemulsion exerted antifungal activity against P. italicum by disrupting cell membrane integrity and permeability; such a nanoemulsion may be used as a potential fungicide substitute for preservation in citrus fruits.

The Protective Effects of Sour Orange (Citrus aurantium L.) Polymethoxyflavones on Mice Irradiation-Induced Intestinal Injury.

Sour orange (Citrus aurantium L.) is one of the biological sources of polymethoxyflavones (PMFs), which are often used to deal with gastrointestinal diseases. The intestine is highly sensitive to irradiation damage. However, limited certain cures have been released for irradiation-induced gastrointestinal injury, and the potentials of sour orange PMFs as radio-resistance agents have not been fully discussed yet. The present study aims to (1) investigate the PMF components in 12 sour orange cultivars, (2) determine the protective effects of PMFs on irradiation-induced intestinal injury by treating mice that received 12 Gy abdominal irradiation with different doses of PMFs and observing the changes in organ indexes and pathological sections and (3) test cytotoxicity of PMFs by CCK-8 method. The results showed that sour orange PMFs appeared to have high intraspecies similarity. Besides, PMFs protected mice from irradiation-induced injury by alleviating body weight loss, reliving organ index changing and maintaining the intestinal structure. Finally, IC50 concentrations to cell line CCD 841 CoN of PMFs and nobiletin were calculated as 42.23 µg/mL and 51.58 µg/mL, respectively. Our study uncovered PMF contents in 12 sour orange materials and determined the protective effects on irradiation-induced intestinal injuries, providing guidance for the utilization of sour orange resources.

Comparative transcriptome and physiological analyses reveal key factors in the tolerance of peach rootstocks to iron deficiency chlorosis.

Iron (Fe) deficiency chlorosis (IDC) is a major nutritional disorder in fruit trees grown on calcareous soils. As a peach rootstock, 'GF677' (Prunus dulcis Miller × P. persica (L.) Batsch) has great tolerance to Fe deficiency, but the molecular mechanisms of 'GF677' that support the process of iron deficiency chlorosis tolerance are still unknown. In this study, the key factors for differential iron deficiency chlorosis tolerance in two contrasting rootstocks (IDC-tolerant: 'GF677', IDC-susceptible: 'Maotao' (P. persica)) were investigated. 'GF677' exhibited greater Fe transfer and accumulation capacities when compared with 'Maotao', and the analysis of photosynthetic pigments, related precursors, and antioxidative enzyme activities further demonstrated that 'GF677' was more tolerant to IDC when compared with 'Maotao'. Furthermore, comparative transcriptome analysis revealed differential expression in many genes involved in iron transport and storage, and in photosynthesis recovery. These results suggest that the greater IDC tolerance of 'GF677' can be attributed to the greater expression of key genes related to specific Fe transporters, defense systems, photosynthetic recovery, and/or special proteins. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03046-6.

Tangeretin prevents obesity by modulating systemic inflammation, fat browning, and gut microbiota in high-fat diet-induced obese C57BL/6 mice.

Obesity and associated comorbidities are closely linked to gut microbiota dysbiosis, energy balance, and chronic inflammation. Tangeretin, a key citrus polymethoxylated flavone (PMF), is abundant in citrus fruits and has preventative and therapeutic effects for numerous diseases. The current study investigated the effects and possible mechanisms of tangeretin supplementation in preventing obesity in high-fat diet (HFD)-fed mice. Treatment of HFD-fed mice with tangeretin potently ameliorated HFD-induced body weight, liver steatosis, glucose intolerance, and insulin resistance. Tangeretin mitigated systemic chronic inflammation by reducing metabolic endotoxemia and inflammation-related gene expression in HFD-fed mice. An increased number of small brown adipocytes possessing multilocular and cytoplasmic lipid droplets and upregulation of thermogenic gene expression were observed after tangeretin treatment. 16S rRNA amplicon sequencing indicated that tangeretin markedly altered the gut microbiota composition (richness and diversity) and reversed 16 operational taxonomic units (OTUs) back to levels seen in mice consuming a normal chow diet (NCD). Notably, tangeretin decreased the ratio of Firmicutes-to-Bacteroidetes and greatly enriched Bacteroides and Lactobacillus. Overall, our results suggest that long-term supplementation with citrus tangeretin ameliorates the phenotype of obesity by improving adipose thermogenesis and reducing systemic inflammation and gut microbiota dysbiosis, which provides a good basis for studying the mechanism of tangeretin's beneficial effects.

Exosomal hsa_circ_0004658 derived from RBPJ overexpressed-macrophages inhibits hepatocellular carcinoma progression via miR-499b-5p/JAM3.

Macrophage-derived exosomes (Mφ-Exo) have multidimensional involvement in tumor initiation, progression, and metastasis, but their regulation in hepatocellular carcinoma (HCC) is not fully understood. RBPJ has been implicated in macrophage activation and plasticity. In this study we assess the role of exosomes derived from RBPJ-overexpressed macrophages (RBPJ+/+ Mφ-Exo) in HCC. The circular RNA (circRNA) profiles in RBPJ+/+ Mφ-Exo and THP-1-like macrophages (WT Mφ)-Exo was evaluated using circRNA microarray. CCK-8, Transwell, and flow cytometry analyses were used to evaluate the function of Mφ-Exo-circRNA on HCC cells. Luciferase reporter assays, RNA immunoprecipitation, and Pearson's correlation analysis were used to confirm interactions. A nude mouse xenograft model was used to further analyze the functional significance of Mφ-Exo-cirRNA in vivo. Our results shown that hsa_circ_0004658 is upregulated in RBPJ+/+ Mφ-Exo compared to WT Mφ-Exo. RBPJ+/+ Mφ-Exo and hsa_circ_0004658 inhibits proliferation and promotes apoptosis in HCC cells, whereas hsa_circ_0004658 knockdown stimulated cell proliferation and migration but restrained apoptosis in vitro and promotes tumor growth in vivo. The effects of RBPJ+/+ Mφ-Exo on HCC cells can be reversed by the hsa_circ_0004658 knockdown. Mechanistic investigations revealed that hsa_circ_0004658 acts as a ceRNA of miR-499b-5p, resulting in the de-repression of JAM3. These results indicate that exosome circRNAs secreted from RBPJ+/+ Mφ inhibits tumor progression through the hsa_circ_0004658/miR-499b-5p/JAM3 pathway and hsa_circ_0004658 may be a diagnostic biomarker and potential target for HCC therapy.

Biosynthesized Gold Nanoparticles Integrated Ointment Base for Repellent Activity Against Aedes aegypti L.

The present study focused on preparing a nano-ointment base integrated with biogenic gold nanoparticles from Artemisia vulgaris L. leaf extract. As prepared, nano-ointment was characterized by using Fourier-transform infrared spectroscopy, and the morphology of the nano-ointment was confirmed through a scanning electron microscope. Initially, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide results showed nano-ointment cytocompatibility at different concentrations (20-200 µg/mL) against L929 cells. The in vitro hemolysis assay also revealed that the nano-ointment is biocompatible. Further studies confirmed that nano-ointment has repellent activity with various concentrations (12.5, 25, 50, 75, and 100 ppm). At 100 ppm concentration, the highest repellent activity was observed at 60-min protection time against the Aedes aegypti L. female mosquitoes. The results indicated that the increasing concentration of nano-ointment prolongs the protection time. Moreover, the outcome of this study provides an alternative nano-ointment to synthetic repellent and insecticides after successful clinical trials. It could be an eco-friendly, safer nano-bio repellent, which can protect from dengue fever mosquitoes.

Tangeretin promotes lifespan associated with insulin/insulin-like growth factor-1 signaling pathway and heat resistance in Caenorhabditis elegans.

Tangeretin is a polymethoxylated flavonoid naturally occurred in citrus fruits with many pharmacological activities, such as anti-inflammatory, antiproliferative, and neuroprotective properties. A previous study reported that tangeretin-enriched orange extract could prolong the lifespan in Caenorhabditis elegans. However, the antiaging effect of tangeretin remains uncertain. In this study, we used the model organism C. elegans to conduct a lifespan test, observed the aging-related functional changes of nematodes, the fluorescence changes of stress-related proteins (DAF-16 and HSP-16.2) and its response to stress assay, and monitored the effect of tangeretin on the mRNA expression levels. The results showed that tangeretin supplementation (30 and 100 µM) extended the mean lifespan, slowed aging-related functional declines, and increased the resistance against heat-shock stress. Furthermore, tangeretin upregulated the mRNA expression of daf-16, hsp-16.2, and hsp-16.49, promoted the nuclear localization of DAF-16, and enhanced the fluorescence intensity of HSP-16.2, while it had no effect on the lifespan of daf-2, age-1, and daf-16 mutants. The current findings suggest that tangeretin can significantly extend the lifespan and enhance heat stress tolerance in an insulin/insulin-like growth factor signaling dependent manner.

[Effects of the combination of tangeretin and whey protein on testosterone and cortisol in sprinters at winter training season].

Objective: To evaluate the regulation efficacy of oral tangeretin on testosterone and cortisol in sprinters at winter training season. Methods: Twenty-four sprinters were paired and randomly divided into experimental group (EG) and control group (CG). During winter training season, EG were treated with 200 mg tangeretin by oral, and CG were treated with placebo for 4 weeks. Blood samples were collected on the first day of each week (T1, T2, T3, T4) and after the intervention (T5) to detect serum levels of testosterone, cortisol, superoxide dismutase (SOD), and adrenocorticotropic hormone (ACTH). The body composition was tested at T1 and T5. Results: After 4 weeks, ①the serum cortisol level of CG was increased, and the serum levels of testosterone and SOD were decreased significantly (P＜0.05). ②In EG, the serum levels of cortisoland ACTH were decreased significantly (P＜0.05, P＜ 0.01), while the serum testosterone level was remained stable, and the level of SOD was increased slightly. ③The muscle mass of EG and CG were increases, but that of EG was increased higher than that of CG. Conclusion: Tangeretin reduces the oxidative stress response that caused by high-intensity exercise during winter training, which maintain the serum testosterone level and inhibit cortisol excessive secretion and promote muscle synthesis.

[Effects of Tangeretin on cortisol stress response induced by high-intensity resistance exercise].

Objective: To investigate the effects of 4-week tangeretin supplementation on cortisol stress response induced by high-intensity resistance exercise. Methods: Twenty-four sprinters were paired and randomly divided into experimental group (EG) and control group (CG). EG orally took supplement with tangeretin (200 mg/day) and CG took placebo for 4 weeks. Before and after the 4-week intervention, all sprinters performed a set of high-intensity resistance exercise (shoulder press, squat, bench press and deadlift, 10 RM, 4 sets per movement) to stimulate their cortisol stress responses. Serum levels of cortisol, adreno-corticotropic hormone (ACTH), superoxide dismutase (SOD), white blood cell count (WBC) and blood glucose were obtained by collecting blood sample before the exercise (PRE), immediately after the exercise (P0), and at 10 (P10), 20 (P20), and 30 minutes (P30) after the exercise. Results: Compared with the same period before the intervention, after the 4-week tangeretin intervention, EXP showed significantly reduced serum cortisol level at PRE (P=0.017), P10 (P=0.010), P20 and P30 (P＜0.05 or P＜0.01), and significantly reduced WBC at PRE, ACTH at P10 (P=0.037) and WBC and ACTH at P30 (P＜0.05). Compared with CTROL, EXP showed significantly lower levels of the serum cortisol at PRE and P10 (P＜0.05), and significantly lower levels of the ACTH (P＜0.05) and WBC (P＜0.01) at P30, and significantly increased level of the SOD activity at P30 (P＜0.05). Conclusion: Tangeretin supplementation can significantly alleviate the cortisol stress response induced by high-intensity resistance exercise, inhibit the excessive secretion of cortisol, enhance antioxidant capacity, accelerate the elimination of inflammation in the body, and promote the recovery of body functions.

An evidence update on the protective mechanism of tangeretin against neuroinflammation based on network pharmacology prediction and transcriptomic analysis.

Although the protective effects of tangeretin on neuroinflammation have been proven in cell and animal experiments, few studies explore its underlying molecular mechanism. In this study, we used the network pharmacology method combined with the transcriptome approach to investigate its underlying anti-inflammatory mechanism in human microglial cells. Based on network pharmacology analysis, four putative target proteins and ten potential pathways were identified. Among them, vascular endothelial growth factor A (VEGFA), epidermal growth factor receptor (EGFR) and the related phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT), the mitogen-activated protein kinase (MAPK), mechanistic target of rapamycin (mTOR) signaling pathway were well-supported by transcriptome data. Meanwhile, transcriptome analysis supplemented two crucial targets: the insulin receptor (InsR) and insulin-like growth factor-I (IGF-1) receptor. Subsequently, VEGFA, EGFR, IGF-1 receptor, and InsR were further verified on the protein level. Taken together, we assumed that tangeretin could exert protective effects on neuroinflammation by decreasing the expression of VEGFA, EGFR, InsR, and IGF-1 receptor in the PI3K-AKT, MAPK, mTOR signaling pathway. More importantly, it is for the first time to show that the anti-neuroinflammatory effects of tangeretin through VEGFA, EGFR, IGF-1 receptor, InsR, and mTOR signaling pathway. These works offer new insight into the anti-neuroinflammatory functions of tangeretin and propose novel information on further anti-inflammatory mechanism studies.