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In this research, we analyzed the protective effects of red-fleshed apple flavonoid extracts (RAFEs) on male reproductive injury induced by busulfan, using both in vitro and in vivo models. In the cell-based experiments, RAFEs significantly improved cell viability and proliferation rates compared to control groups. Similarly, in vivo testing with male mice showed that RAFEs and whole apple flavonoid extracts (WAFEs) enhanced various biochemical and liver function-related indicators in the testes; however, RAFEs demonstrated superior efficacy in mitigating testicular damage. Through immunohistochemistry, qRT-PCR, and Western blotting, we found that RAFEs notably enhanced the expression of spermatogenesis-related genes. Moreover, RAFEs increased the expression of oxidative stress- and apoptosis-related genes, thereby effectively reducing oxidative damage in the testes. These findings highlight the potential of RAFEs as natural agents for the prevention and treatment of male reproductive injury, paving the way for future research and potential therapeutic applications.

Red-fleshedapples are preferredbecause of their high content of phenolics and antioxidants in peel and pulp. Herein, we evaluated the mechanisms of apple peel polyphenolic extracts (APP) and apple flesh polyphenolic extracts (AFP) from the new red-fleshed apple in inhibiting cell proliferation and inducing apoptosis on human breast cancer MDA-MB-231 cells. The antiproliferative activities were determined by the CCK8 assay. The expression of proteins was determined using Western blot. We found that the content of polyphenols and flavonoids in APP was significantly higher than that in AFP, and 14 main phenolic compounds in APP and AFP were quantified using UPLC-MS/MS techniques. Besides, the significant inhibition effects of APP and AFP were achieved through Akt pathway by inducing apoptosis (significantly upregulating reactive oxygen species [ROS] levels, and downregulating expression of pAkt, pBad, Bcl-2, promoting Cytochrome c release, activating Cle-Caspase 9, and inducing expressions of Cle-Caspase 3 and Cle-PARP), and inducing G0/G1 cell cycle arrest (increased expressions of p-p53 and p21 and decreased expressions of PCNA and Cyclin D1). And the inhibition effect of APP was stronger than that of AFP. These results suggest that AFP and APP may be excellent sources of natural chemicals for treating triple-negative breast cancer MDA-MB-231 cells. PRACTICAL APPLICATION: The effects of antiproliferation of phenolic extracts from red-fleshed apple peels and flesh on human breast cancer MDA-MB-231 cells were evaluated. The data may clarify the functional parts of red-fleshed apple and provide some basis for scientific researchers and consumers to recognize and exploit red-fleshed apple., (© 2021 Institute of Food Technologists®.)

Apple (Malus domestica Borkh.) is an important fruit tree species worldwide. Apple fruits are favored by consumers because of their antioxidative, anti-inflammatory, and antitumor effects as well as their protective effects against cardiovascular diseases and other health benefits. There is considerable interest in red-fleshed apple fruits among breeders because of their high flavonoid and anthocyanin contents. However, the flavonoids extracted from red-fleshed apple fruits must still be functionally characterized, especially regarding their protective effects against certain pathologies. In this study, the flavonoid components and contents in the extracts prepared from red-fleshed apple cultivar "Meihong" were determined. Additionally, the in vitro antioxidant activities and protective effects of the extracts against CCl 4 -induced acute liver injury were investigated. The red-fleshed apple flesh flavonoid extract (RAFF) exhibited strong in vitro antioxidant activities. Compared with the model control mice treated with CCl 4 , the mice pretreated with high (800 mg/kg·bw), middle (400 mg/kg·bw), and low (200 mg/kg·bw) RAFF doses had significantly lower CCl 4 -induced serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase activities. Moreover, the RAFF pretreatment also significantly decreased the liver malondialdehyde activity and prevented the CCl 4 -induced decrease in liver superoxide dismutase, glutathione peroxidase, catalase, and reduced glutathione levels. Furthermore, a histopathological examination revealed that RAFF inhibited the inflammatory cell infiltration and cell boundary loss caused by CCl 4 in the liver. Thus, RAFF is a natural antioxidant with significant antioxidative activities and liver protective effects. The results of this study may be relevant for enhancing the application of the red-fleshed apple fruit extract as a food additive. PRACTICAL APPLICATION: We took the self-selected red-fleshed apple cultivar "Meihong" as the unique research material, and the active ingredients of its flavonoid extract, in vitro antioxidant activity and hepatoprotective effect were analyzed. It is of great significance to promote the development of the red-fleshed apple industry, and also provides an important reference for the development of natural antioxidants., (© 2020 Institute of Food Technologists®.)

In this research, we compared the phenotypical characters, total anthocyanins content, total phenols content, and antioxidant activity of red-fleshed apple cultivars 'XJ4', 'QN-5', 'DH' and 'HX1' at three fruit developmental stages. A further flavonoids metabolites study was conducted in 'XJ4' and 'DH'. We found broader variation of total anthocyanins content in the peel of the four cultivars, which might result in larger differences of free radicals scavenging rate. The most significant difference in fruit phenotype, anthocyanins content, and DPPH scavenging rate was observed between 'XJ4' and 'DH' at mature stage. Therefore, the flavonoids metabolites of 'XJ4' and 'DH' at mature stage were compared to unveil the details of anthocyanins compounds. The unique compounds pelargonidin 3- O -β-d-glucoside and cyanidin-3- O -malonylhexoside were detected only in peel and flesh of 'XJ4' but not in 'DH', which might contribute to the purple peel and dark-red flesh color of 'XJ4'. Significantly decreased upstream metabolites in the early biosynthetic genes regulated domain were found only in 'XJ4' peel but not in the flesh. This might explain why the anthocyanins content in 'XJ4' peel was decreased largely at the mature stage. Taken together, our findings will give some insight into the metabolites study in flavonoid biosynthetic pathway of red-fleshed apple.

Background: Red-fleshed apples are a great source of natural colorants and functional food ingredients because of their high anthocyanin content. Generally, anthocyanins are highly unstable after extraction, which limits their wide applications in the food and pharmaceutical industries. This study was aimed at investigating the effects of combining copigmentation with encapsulation on the stability of anthocyanins from red-fleshed apples. In this study, red-fleshed apple anthocyanins were copigmented with caffeic acid, and then the copigmented complexes were encapsulated using gum arabic and maltodextrin using spray drying and freeze drying., Results: All anthocyanin microcapsules had high encapsulation efficiencies ranging from 93.84 to 96.85% with mean hydrodynamic diameter smaller than 350 nm. After heating at 80 °C for 2 h, the dispersions of microencapsulated anthocyanins with copigments exhibited the highest absorbance values at λ max (515 nm) (P < 0.05). Light stability experiments demonstrated that the half-life of the red-fleshed apple anthocyanins increased from 5 to 12 days after being treated with copigmentation and encapsulation. The drying methods (spray/freeze drying) did not significantly influence the stability of the microencapsulated anthocyanins., Conclusions: Applying copigmentation and spray-drying encapsulation in tandem has great potential for enhancing the stability of red-fleshed apple anthocyanins. Thus, such anthocyanins with enhanced stability may be increasingly used in the food and pharmaceutical industries as value-added natural food pigments. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)

Red-flesh apples are known as functional fruits because of their rich anthocyanin. The anthocyanin content of the red flesh apple cultivar 'QN-5' we bred can reach 361 mg·kg -1 (FW), and showed higher scavenging capacity to DPPH radicals, hydroxyl radicals, and superoxide anion radicals, with scavenging rates of 80.0%, 54.0%, and 43.3%, respectively. We used this particular anthocyanin-rich 'QN-5' apple as material to examine how nanocrystallization affects the antixodiant effect of anthocyanin. The anthocyanin extract was encapsulated with biocompatible zein to form zein-anthocyanin nanoparticles (ZANPs). Transmission electron microscopy (TEM) scanning showed that ZANPs had a regular spherical shape with an average diameter size of 50-60nm. When the ratio of the zein and the anthocyanin was 1:0.5, the results suggested that the encapsulation efficiency (EE) of the ZANPs could reach as high as 92.8%, and that scavenging rate for DPPH radicals was increased from 87.1% to 97.2% compared to the non-nanocrystallized anthocyanin extract. Interestingly, treatment under alkaline conditions (pH 9.0), high temperature (90 °C), and a storage time of 7 days could decrease the scavenging capacity of the ZANPs for DPPH radicals, but this scavenging capacity loss for ZANPs was significantly lower than that observed in the non-nanocrystallized anthocyanin, suggesting the higher stability of ZANPs is caused by encapsulation. These results would provide a theoretical basis for the application of the anthocyanin in scavenging free radicals under stress conditions.