Lactation-promoting ingredients of Hemerocallis citrina Borani and the corresponding mechanisms.

Hemerocallis citrina Borani is a traditional folk food used to promote the lactation of postpartum mothers in China; however, the active ingredients and corresponding mechanisms are still unknown. In this study, the lactogenic effect of alcoholic and aqueous extracts of H. citrina was primarily evaluated, and the aqueous extract (1,000 and 2,000 mg/kg) displayed significant lactation-promoting effects. Three eluates of the aqueous extract (0%, 30%, and 50%HCW) were further evaluated for their lactogenic effect, and 30% and 50% HCW showed significant lactation-promoting activity. Nineteen ingredients, including those with a high content of rutin and isoquercetin, were then identified from 30% and 50%HCW using the ultra-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS) method. Finally, the lactogenic effect of rutin and isoquercetin was evaluated, and both compounds displayed significant lactation-promoting activity. The mechanisms relative to the lactation-promoting active ingredients for H. citrina extracts and compounds are to stimulate the release of prolactin (PRL) and progesterone (P), as well as to induce the expression of prolactin receptor (PRLR) and improve the morphology of mammary tissue. This study first clarified the lactation-promoting active ingredients of H. citrina and the corresponding mechanisms, which provide a new insight into the new lactation-promoting drug and promote the high-value utilization of H. citrina resources.

A novel protein extracted from Hemerocallis citrina Borani inhibits hepatocellular carcinoma cell proliferation by regulating mitochondria-dependent apoptosis and aerobic glycolysis.

Hemerocallis citrina Borani is a commonly consumed food in Asia and possesses many biologically active ingredients. In this study, a protein named Hemerocallis citrina Borani protein (HcBP) was purified using ammonium sulfate fractionation and anion exchange chromatography. Protease assays revealed that HcBP has peroxidase activity. Meanwhile, the UV absorption spectrum showed that HcBP contains heme. Notably, HcBP showed significant inhibitory effects on human hepatoma cancer cell proliferation. Mechanism investigations indicated that HcBP treatment resulted in overproduction of reactive oxygen species (ROS) and induced mitochondria-dependent apoptosis in human hepatoma cancer cells. Furthermore, we found HcBP not only downregulated pyruvate kinase M2 (PKM2) activity but also decreased the expression and nuclear levels of PKM2. The inhibition of PKM2 led to the downregulation of GLUT1, LDHA and PDK, and thus caused the suppression of glycolysis. In summary, our results suggested that HcBP has potential anti-hepatocellular carcinoma activity.

Freeze-dried powder of daylily bud improves bromocriptine-induced lactation disorder in rats via JAK2/STAT5 pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Milk deficiency is a prevalent problem in the world. Daylily (Hemerocallis citrina Borani), called the Chinese mother flower, is a traditional vegetable and is believed to possess a galactagogue effect in China. Flavonoids and phenols are considered as the active ingredients of daylily to promote lactation and improve depression. AIM OF THE STUDY: The aim of this study was to investigate the prolactin effects of freeze-dried powder of flower buds of H. citrina Baroni in rat and its action mechanisms. MATERIALS AND METHODS: The chemical constituents of flower buds of H. citrina Baroni treated by different drying techniques were analyzed by ultrahigh pressure liquid chromatography-mass spectrometry. Sprague-Dawley (SD) rat model induced by bromocriptine was used to evaluate the effect of freeze-dried powder of daylily buds on promoting lactation. Network pharmacology method, ELISA, qPCR, and Western blot were used to clarify the action mechanisms. RESULTS: We detected 657 compounds in daylily buds. The relative contents of total flavonoids and phenols in freeze-dried samples were higher than those in dried ones. Bromocriptine, as a dopamine receptor agonist, can significantly inhibit prolactin in rats. Daylily buds can restore the levels of prolactin, progesterone and estradiol depressed by bromocriptine, effectively improve the milk production of the rat, and promote the repair of rat mammary gland tissue. We analyzed the relationship between the chemical components of daylily buds and the genes related to lactation with network pharmacology method, revealing that flavonoids and phenols may be the active components that promoted milk production via JAK2/STAT5 pathway, which was confirmed by the results of qPCR and Western blot. Daylily buds can increase the mRNA expression of PRLR, CSN2, LALBA and FASN and the protein expression of PRLR, JAK2 and STAT5. CONCLUSION: Daylily buds can improve the insufficient lactation of rats induced by bromocriptine through PRLR/JAK2/STAT5 pathway, and the freeze-dried processing method may better retain the active components of flavonoids and phenols that promote milk in daylily.

Polysaccharide from Hemerocallis citrina Borani by subcritical water with different temperatures and investigation of its physicochemical properties and antioxidant activity.

Hemerocallis citrina Borani as a low-cost vegetable, has various health benefits. However, the industry of H. citrina Borani is in the state of primary processing, with poor economic benefits. This study aimed to investigate the physicochemical properties, and the antioxidant activity of H. citrina Borani polysaccharide (HCBP) using subcritical water extraction (SWE) at different temperatures, to expand the value of H. citrina Borani. HCBP mainly composed of nine monosaccharides (glucose, galactose, rhamnose, fucose, mannose, arabinose, xylose, galacturonic acid, and glucuronic acid), among which the content of neutral sugar was higher and uronic acid was lower. HCBP contained glycosidic bond of ß-configurations and trace quantities protein. The molecular weight of HCBP decreased with increasing temperature. Shear thinning occurred in HCBP with the increase of shear rate (0.01-1 s-1), and the apparent viscosity of HCBP decreased at higher temperature (150-160°C) with the increase continuously of shear rate (1-10 s-1), but almost remained constant at lower temperature (130-140°C). Scanning electron microscope showed that HCBP had rough surface, loose structure, obvious particle gap, and irregular shape. In addition, HCBP extracted at 160°C had strong FRAP activity, and HCBP extracted at 130 and 140°C had better ABTS radical scavenging activity. This study suggests that HCBP extracted by SWE could provide a cheap raw material as food thickening agent and natural antioxidants.

Polysaccharide from Hemerocallis citrina Borani by subcritical water: Bioactivity, purification, characterization, and anti-diabetic effects in T2DM rats.

Hemerocallis citrina Borani (daylily) has various health benefits. However, the structural characterization and hypoglycemic effects of its polysaccharide remain unclear. Here, we first report for the first time, the effects of subcritical water extraction temperature on bioactivity of Hemerocallis citrina Borani polysaccharide (HCBP). HCBP extracted at 160 °C had better scavenging ability of free-radical and pancreatic lipase inhibition. This study aimed to investigate the structural characterization and anti-diabetic effects of aforementioned HCBP. HCBP was separated into three fractions using a DEAE-Sepharose Fast Flow. HCBP-1 was the major component with lower molecular weight, HCBP-2 was the least abundant component, HCBP-3 was the component with higher uronic acid and molecular weight. HCBP treatment significantly decreased the concentrations of fasting blood glucose, insulin, total cholesterol, triglyceride, and low density lipoprotein, as well as improved glucose intolerance and insulin resistance in type 2 diabetes mellitus (T2DM) rats. HCBP also protected the kidneys and pancreatic organs by histopathology and immunohistochemical analyses in T2DM rats. In addition, HCBP significantly increased the antioxidant enzymes activities and decreased the level of malonaldehyde to alleviated the oxidative stress injury. The results indicate HCBP extracted by subcritical water is promising functional food ingredients and could be used in T2DM treatment.

Systematic identification metabolites of Hemerocallis citrina Borani by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry combined with a screening method.

Hemerocallis citrina Borani is an important crop and its flower buds are widely consumed in East Asian areas as a vegetable, as well as in traditional Chinese medicine, due to its health-promoting properties. Metabolites present in plant-derived foods or medicines are in part responsible for their desirable flavor profiles and health benefits. Nevertheless, detailed information about these compounds in H. citrina is scarce. Therefore, this study aimed to investigate the metabolites by high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). In this study, a total of 144 compounds, including 14 amides, 25 polyphenols, 44 flavonols, 35 anthraquinones, 15 naphthols, and 11 other components, were detected by the established screening method and were identified by their precise m/z values, characteristic tandem mass spectrometry (MS/MS) data and fragmentation pathways of references, 111 of which were reported in this plant for the first time. The distribution of identified ingredients in different parts of H. citrina was determined. Interestingly, colchicine, which had been reported as a toxic compound in the fresh flower buds in previous studies and various news reports, was not found. This work marks the first comprehensive study of metabolites from commercial flower buds and different parts of H. citrina.