Effects of Humulus lupulus L. and its active ingredient xanthohumol on preventing glucocorticoid-induced osteoporosis / 药学实践杂志

Objective To explore the effects of Humulus lupulus L. extract (HLE) and xanthohumol (XN) on preventing glucocorticoid-induced osteoporosis (GIOP). Methods The GIOP model was established by intraperitoneal injection of dexamethasone (DEX). Bone microstructure, bone mineral density and serum biochemical indexes were evaluated by Micro-CT and ELISA kits. The levels of cells proliferation and ALP activity, and the expression of bone formation related proteins were assayed with primary osteoblasts injured by DEX. Results HLE and XN significantly alleviated the bone microstructure damage, enhanced the bone mineral density, and improved the trabecular parameters in GIOP mice. In vitro experiments showed that HLE and XN can prevent bone loss not only by improving cell proliferation and ALP activity, but also through increasing the expression of bone γ-glutamic acid-containing proteins (BGP), bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (Runx-2). Conclusion This study confirmed that HLE and XN had anti-GIOP effects for the first time. It provides a new resource for the development of anti-osteoporosis medications.

Hop-derived Xanthohumol Induces HL-60 Leukemia Cells Death

Background : Acute promyelocytic leukemia (APL) affects both kids and adults, however it is more prevalent in younger population. Although APL has a favorable prognostic, patients that relapse often do not respond positively to additional chemotherapy. Therefore, there is a need to further identify ways to overcome these challenges. Hypothesis: In this study, we examined antileukemic effects of xanthohumol (XN), a prenylated flav onoid derived from hops ( Humulus lupulus L ), on human promyelocytic HL - 60 cells. Materials and Methods : HL - 60 cells were exposed to different concentrations of XN (?M) for 24 h. Cell viability, cell morphology, chromatin condensation, cPARP - 1 level, and caspase - 3 activation, and the expression of p21 WAF1/Cip1 were analyzed. Results : XN reduced HL - 60 cell viability in a dose - dependent manner. XN induced a dose - dependent morphological changes including cell shrinkage and b lebbing , and significantly increased the number of cells with condensed chromatin. XN significantly increased the level of cPARP - 1, active caspase - 3, and the expression of p21WAF/CIP mRNA. Conclusion : These data indicate that XN induces HL - 60 cell death by regula ting cell cycle progression and apoptosis. This study suggests that XN may have antileukemic preventive effects.

Cytotoxic Effects of Xanthohumol and Its Combination with Cisplatin on Human Metastatic Lung Cancer H1299 Cells

Aims:Non-small cell lung cancer (NSCLC) accounts for high lung cancer death that is mostly associated with advanced disease stage at diagnosis and resistance to chemotherapy. In the present study, we investigated whether xanthohumol, a prenylated flavonoid of hop plant, induces metastatic lung cancer H1299 cell death, and whether in combination with cisplatin there are additive effects. Methodology:H1299 cells were grown and treated with xanthohumol (6.25, 12.5, or 25 μM), cisplatin (12.5, 25, or 50 μM) and the combination of cisplatin and xanthohumol for 24 h. Cell viability, cell morphology, chromatin condensation, ɣH2AX, cPARP-1, capsase-3, p21WAF1/CIP1and p14ARFgenes were analyzed Results:Xanthohumol, cisplatin, and the combination of cisplatin and xanthohumol inhibited H1299 cells viability. Cisplatin growth inhibitory effects were potentiated by xanthohumol. Xanthohumol induced chromatin condensation and apoptosis and potentiated cisplatin’s effect vs cisplatin alone. Further investigation of growth inhibitory effects, xanthohumol alone induced γH2AX foci formation and the combination potentiated γH2AX foci formation. Cisplatin, xanthohumol at 25 μM, and the combination of cisplatin and xanthohumol at 6.25 and 12.5 μM increased cPARP-1 level. Active caspase-3 was increased by cisplatin, 12.5 μM of xanthohumol, and the combination of xanthohumol and cisplatin. Xanthohumol at 6.25 or 12.5 μM potentiated cisplatin effect on active caspase-3 and cPARP-1, respectively. Xanthohumol at 25 μM significtly induced the expression cell cycle control genes p21WAF1/CIP1and p14ARF. These results indicate that xanthohumol inhibits proliferation of H1299 cells and induces cell death through cleavage of PARP-1 and activation of caspase-3. The combination of cisplatin and xanthohumol potentiated cytotoxic effects of each other compound.Conclusion:The present study suggests that xanthohumol poses apoptotic effects and potentiates cisplatin’s growth inhibitory effects on metastatic lung cancer cells

Anti-osteoporotic study on xanthohumol, an active ingredient in Humulus lupulus L / 药学实践杂志

Objective To evaluate the anti-osteoporotic effect of xanthohumol(XN)in animal and osteoblast.Methods The anti-osteoporotic study on XN was performed with ovariectomized mice model.Serum biochemical indexes,bone mineral density(BMD)and bone histomorphology were measured using Elisa kits and Micro-CT analysis.In vitro test,the effect of XN on osteoblastic proliferation,differentiation and mineralization were assayed.The expression of protein related to bone for-mation was measured by Western blot analysis.Results In vivo experiments,XN significantly increased the estrogen level, reduced the high bone turnover rate,improved the microenvironment and BMD in ovariectomized mice.In vitro experiments, XN protected bone loss not only by promoting osteoblastic proliferation,ALP activity and bone mineralization,but also through increasing the expression of osteopontin(OPN),bone sialoprotein(BSP)and bone morphogenetic protein-2(BMP-2). Conclusion This is the first report to confirm that XN has anti-osteoporotic effect,which provides a new approach for the clin-ical treatment of osteoporosis.

Analysis of flavonoids contents of hops / 药学实践杂志

Objective To determine the contents of total flavonoids and xanthohumol in hop from 29 different countries and regions .Methods Rutin colorimetric method was used to determine the content of total flavonoids .HPLC-UV method was established for the determination of xanthohumol in hops .HPLC method was performed by Dikma Technologies Diamonsil C18 (250 mm × 4 .6 mm ,5 μm ) column with mobile phase acetonitrile-1％ glacial acetic acid solution at the flow rate of 1 .0 ml/min .The column temperature was 25 ℃ .The detection wavelength was 370 nm .Results The equation of linear re-gression of total flavonoids was A=30 .345C+0 .0168 ,r=0 .9999 .The equation of linear regression of xanthohumol was A=55446 C+9040 .5 ,r=0 .9999 .Their linear ranges were respectively 20 .2-404 .0 μg/ml ,2 .152-43 .040 μg/ml ,which indica-ted a good linear relationship .The RSDs of precision and repeatability were less than 2％ .The average recoveries of flavonoids and xanthohumol were respectively 102 .71％ and 100 .21％ .Conclusion The contents of total flavonoids and xanthohumol in different hops varieties are significantly different and the import hops was better than the domestic hops in this study .

Stability of Active Constituents of Hops (Humulus lupulus) Strobiles and their Ethanolic Extracts during Storage.

Aims: The purpose of this study was to evaluate the stability of three major active constituents (humulones, lupulones and xanthohumol) in dried hops (Humulus lupulus) strobiles (whole and ground) as well as their ethanolic extracts during storage. Methodology: A comparative study of humulones, lupulones and xanthohumol levels of H. lupulus strobiles during storage was carried out. Dried whole strobiles and cryogenically ground dried strobiles stored at -15ºC as well as ethanol extracts of the strobiles prepared using different ethanol concentrations (10%, 30%, 50%, 70%, and 95%) and stored at room temperature, were analyzed by HPLC to quantify each constituent. These hops samples were analyzed immediately after preparation, and then one year and two years later to determine the concentrations of the constituents. Results: HPLC analysis indicated that the amount of all three constituents in the ground strobiles and in the ethanol extracts decreased gradually during the storage period. The 10% and 30% ethanol extracts had very low amounts of constituents initially and were practically devoid of constituents at the end of two years. The 50% ethanol extract contained considerable amounts of humulones and xanthohumol, and low levels of lupulones initially, but lost substantial amounts over time. The 70% and 95% ethanol extracts showed higher levels of all three constituents, while the 95% H. lupulus ethanol extract contained the highest constituent levels throughout the experimental period. The ethanol content of the extract had a direct correlation to the constituent levels; the higher the ethanol level, the higher the initial and subsequent constituent levels. Conclusion: Both dried hops and ethanol extracts lose active components over storage time. When preparing extracts, at least 70% ethanol is necessary to extract the highest levels of three bioactive constituents and to retain them over a two-year period. Ethanol concentration is a critical factor to be considered in hops extraction process.

Xanthohumol inhibits cellular proliferation in a breast cancer cell line (MDA‑MB231) through an intrinsic mitochondrial‑dependent pathway.

AIMS: Xanthohumol isolated from hops has been reported to exhibit anticancer effects in diverse human cancers. However, its effect on breast cancer has not yet been clearly defined. The purpose of this study was to investigate the effects of xanthohumol on breast cancer cell proliferation. MATERIALS AND METHODS: After treatment with 5 µM, 10 µM, and 20 µM xanthohumol for 48 h, cells from the human breast cancer cell line MDA‑MB‑231 were studied using colony assay, flow cytometry, and western blotting. RESULTS: The survival rate of the MDA‑MB231 cells treated with 10 µM and 20 µM xanthohumol for 48 h decreased significantly by 64.7 ± 1.8% and 40.1 ± 1.8%, respectively. The numbers of SubG0/G1 cells in the group treated with 10 µM and 20 µM xanthohumol increased significantly to 11.3 ± 0.2 and 18.4 ± 0.1, respectively. A ladder pattern of DNA fragmentation was also observed. Xanthohumol increased the expression of Bax in the mitochondria, which correspondingly decreased in the cytoplasm. The activity of caspase‑3 and caspase‑9 was shown to increase significantly in the treated groups but not in the control group. CONCLUSIONS: Xanthohumol inhibited the proliferation of MDA‑MB‑231 cells through a mitochondria‑ and caspase‑dependent apoptotic pathway. This result suggests that xanthohumol might serve as a novel therapeutic drug for breast cancer.

Effect of xanthohumol on melanogenesis in B16 melanoma cells

Xanthohumol (XH), the principal prenylflavonoid of the hop plant (Humulus lupulus L.), dose-dependently inhibited isobutylmethylxanthine (IBMX)-induced melanogenesis in B16 melanoma cells, with little cytotoxicity at the effective concentrations. Decreased melanin content was accompanied by reduced tyrosinase enzyme activity, protein and mRNA expression. The levels of tyrosinase-related protein 1 and 2 mRNAs were decreased by XH. XH also inhibited alpha-melanocyte stimulating hormone- or forskolin-induced increases in melanogenesis, suggesting an action on the cAMP-dependent melanogenic pathway. XH downregulated the protein and mRNA expression of microphthalmia-associated transcription factor (MITF), a master transcriptional regulator of key melanogenic enzymes. These results suggest that XH might act as a hypo-pigmenting agent through the downregulation of MITF in the cAMP-dependent melanogenic pathway.