Chrysin alleviates cerebral ischemia-reperfusion injury by inhibiting ferroptosis in rats / 中国中药杂志

The purpose of this study is to investigate whether chrysin reduces cerebral ischemia-reperfusion injury(CIRI) by inhi-biting ferroptosis in rats. Male SD rats were randomly divided into a sham group, a model group, high-, medium-, and low-dose chrysin groups(200, 100, and 50 mg·kg~(-1)), and a positive drug group(Ginaton, 21.6 mg·kg~(-1)). The CIRI model was induced in rats by transient middle cerebral artery occlusion(tMCAO). The indexes were evaluated and the samples were taken 24 h after the operation. The neurological deficit score was used to detect neurological function. The 2,3,5-triphenyl tetrazolium chloride(TTC) staining was used to detect the cerebral infarction area. Hematoxylin-eosin(HE) staining and Nissl staining were used to observe the morphological structure of brain tissues. Prussian blue staining was used to observe the iron accumulation in the brain. Total iron, lipid pero-xide, and malondialdehyde in serum and brain tissues were detected by biochemical reagents. Real-time quantitative polymerase chain reaction(RT-qPCR), immunohistochemistry, and Western blot were used to detect mRNA and protein expression of solute carrier fa-mily 7 member 11(SLC7A11), transferrin receptor 1(TFR1), glutathione peroxidase 4(GPX4), acyl-CoA synthetase long chain family member 4(ACSL4), and prostaglandin-endoperoxide synthase 2(PTGS2) in brain tissues. Compared with the model group, the groups with drug intervention showed restored neurological function, decreased cerebral infarction rate, and alleviated pathological changes. The low-dose chrysin group was selected as the optimal dosing group. Compared with the model group, the chrysin groups showed reduced content of total iron, lipid peroxide, and malondialdehyde in brain tissues and serum, increased mRNA and protein expression levels of SLC7A11 and GPX4, and decreased mRNA and protein expression levels of TFR1, PTGS2, and ACSL4. Chrysin may regulate iron metabolism via regulating the related targets of ferroptosis and inhibit neuronal ferroptosis induced by CIRI.

Investigation on improvement effects and mechanism of 7-hydroxyethyl chrysin on PC12 cell injury induced by hypobaric hypoxia / 中国药房

OBJECTIVE To investigate the improvement effects and possible mechanism of 7-hydroxyethyl chrysin （7-HEC） on PC12 cell injury induced by hypobaric hypoxia. METHODS The rat adrenal pheochromocytoma cell line PC12 was cultured under low-pressure hypoxia （5%CO2， 94%N2， 1%O2， 54 004 Pa） to investigate the different concentrations of 7-HEC （100， 10， 1， 0.1， 0.01 μmol/L） on the survival rate of hypoxic cells； the effects of 7-HEC（1 μmol/L） on the contents of lactate dehydrogenase （LDH） and malondialdehyde （MDA）， superoxide dismutase （SOD） activity， apoptotic rate， cell cycle， and the expressions of cleaved caspase-3， Bcl-2 and Bax were detected. RESULTS Compared with control group， the survival rate of cells in hypobaric hypoxia group was decreased significantly （P＜0.01）； 10， 1， 0.1 μmol/L 7-HEC could reverse the decrease of cell survival rate caused by hypobaric hypoxia （P＜0.05 or P＜0.01）. Compared with control group， LDH content in supernatant， MDA content in cells， apoptotic rate， the proportion of cells at G1 stage and the protein expression of cleaved caspase-3 were increased significantly in hypobaric hypoxia group， while SOD activity in cells， the proportion of cells at S stage and G2 stage and Bcl-2/Bax ratio were decreased significantly （P＜0.05 or P＜0.01）. Compared with hypobaric hypoxia group， the contents of LDH and MDA， apoptotic rate， the proportion of cells at G1 stage and the expression of cleaved caspase-3 in 7-HEC group were decreased significantly， while SOD activity， the proportion of cells at G2 stage and Bcl-2/Bax ratio were increased significantly （P＜ 0.01）. CONCLUSIONS 7-HEC can significantly increase the survival rate of hypobaric hypoxia cells， reduce the LDH content in supernatant， improve cell cycle arrest， and reduce the rate of apoptosis. Its improvement effects on hypobaric hypoxia cell injury may be related to the inhibition of caspase-3/Bax/Bcl-2 pathway activation.

Protective effect of chrysin regulates AMPK-NLRP3 signaling mediated pyroptosis to alleviate hepatic fibrosis / 药学学报

This study investigated the protective effect of chrysin on hepatic fibrosis by regulating AMP-activated kinase (AMPK)-NOD-like receptor protein 3 (NLRP3) mediated pyroptosis pathway. The hepatic fibrosis model of mice was established by thioacetamide (TAA) in vivo. Except the control and chrysin alone groups, the mice were injected intraperitoneally with TAA at 100 mg·kg-1, three times per week for the first week. From the 2nd to 5th week, mice were injected intraperitoneally with TAA at 200 mg·kg-1, three times per week for the next 4 weeks. Chrysin groups were intragastrically administrated once per day to 5th week. The histopathological changes were detected by HE and Masson staining. The levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assessed by the kits. All animal experiments were approved by the Medical Ethics Committee of Affiliated Zhongshan Hospital of Dalian University (DWLL2019060). LX-2 cells were stimulated by (transforming growth factor-β, TGF-β) in vitro. The protein expressions of AMPKα, p-AMPKα, NLRP3, cysteinyl aspartate specific proteinase-1 (caspase-1), gasdermin D (GSDMD) were detected by Western blot, and the mRNA levels of collagen-Ι, α-smooth muscle actin (α-SMA), interleukin-1β (IL-1β), IL-18, caspase-1, GSDMD were analysis by reverse transcription-polymerase chain reaction (RT-PCR). Chrysin attenuated the increases in serum AST and ALT levels in the TAA group, while significantly improved the changes of liver morphology, reduced liver tissue inflammatory cell infiltration and inhibited collagens deposition. Compared with TAA group, chrysin effectively activated AMPKα phosphorylation and inhibited hepatic NLRP3 inflammasome activation. Additionally, the protein expressions and mRNA levels of IL-1β, IL-18, caspase-1 and GSDMD in chrysin groups were decreased. Chrysin inhibited the expressions of collagen-Ι and α-SMA, enhanced the phosphorylation of AMPKα, and decreased the expressions of NLRP3 and GSDMD. Therefore, chrysin may inhibit inflammatory injury and pyroptosis possibly by activating AMPK and inhibiting NLRP3 inflammasome to alleviate hepatic fibrosis.

Chrysin-Enhanced Cytotoxicity of 5-Fluorouracil-Based Chemotherapy for Colorectal Cancer in Mice: Investigating its Effects on Cyclooxygenase-2 Expression

Abstract 5-fluorouracil (5-FU) has been recognized as an effective medication used to treat colorectal cancer (CRC); however, its administration is facing limitations due to some complications reported. It is also generally accepted that combination therapy is among strategies to improve chemotherapy efficiency. Therefore, chrysin, with its anticancer effects, in combination with 5-FU was investigated in the present study. Azoxymethane (AOM) as a carcinogenic substance along with dextran sodium sulfate (DSS) was additionally utilized to induce CRC in mice. The anticancer effects of chrysin were then evaluated using aberrant crypt foci (ACF) counting and percentage of pathologic lesions in epithelial tissues from distal colon. In this study, cyclooxygenase (COX-2) protein expression was correspondingly explored through immunohistochemistry (IHC). The results revealed that chrysin alone or in combination with 5-FU could decrease ACF counting and percentage of pathologic lesions in comparison with AOM (p<0.05). Moreover, the combination of chrysin (at a dose of 50 mg/kg) with 5-FU reduced COX-2 expression compared with 5-FU alone (p<0.001) or 5-FU in combination with chrysin at a dose of 100 mg/kg (p<0.05). Furthermore, the combined chrysin boosted 5-FU efficiency, so it was suggested as an auxiliary therapy for CRC.

Trypanosuppressive effects of Kolaviron may be associated with down regulation of Trypanothione reductase in Trypanosoma congolense infection

@#Trypanothione reductase is a key enzyme that upholds the redox balance in hemoflagellate protozoan parasites such as T. congolense. This study aims at unraveling the potency of Kolaviron against trypanothione reductase in T. congolense infection using Chrysin as standard. The experiment was performed using three different approaches; in silico, in vitro and in vivo. Kolaviron and Chrysin were docked against trypanothione reductase, revealing binding energies (-9.3 and -9.0 kcal/mol) and Ki of 0.211μM and 0.151μM at the active site of trypanothione reductase as evident from the observed strong hydrophobic/hydrogen bond interactions. Parasitized blood was used for parasite isolation and trypanothione reductase activity assay using standard protocol. Real-time PCR (qPCR) assay was implored to monitor expression of trypanothione reductase using primers targeting the 177-bp repeat satellite DNA in T. congolense with SYBR Green to monitor product accumulation. Kolaviron showed IC50 values of 2.64μg/ml with % inhibition of 66.78 compared with Chrysin with IC50 values of 1.86μg/ml and % inhibition of 53.80. In vivo studies following the administration of these compounds orally after 7 days post inoculation resulted in % inhibition of Chrysin (57.67) and Kolaviron (46.90). Equally, Kolaviron relative to Chrysin down regulated the expression trypanothione reductase gene by 1.352 as compared to 3.530 of the infected group, in clear agreement with the earlier inhibition observed at the fine type level. Overall, the findings may have unraveled the Kolaviron potency against Trypanosoma congolense infection in rats.

Chrysin serves as a novel inhibitor of DGKα/FAK interaction to suppress the malignancy of esophageal squamous cell carcinoma (ESCC)

@#Among current novel druggable targets, protein–protein interactions (PPIs) are of considerable and growing interest. Diacylglycerol kinase α (DGKα) interacts with focal adhesion kinase (FAK) band 4.1-ezrin-radixin-moesin (FERM) domain to induce the phosphorylation of FAK Tyr397 site and promotes the malignant progression of esophageal squamous cell carcinoma (ESCC) cells. Chrysin is a multi-functional bioactive flavonoid, and possesses potential anticancer activity, whereas little is known about the anticancer activity and exact molecular mechanisms of chrysin in ESCC treatment. In this study, we found that chrysin significantly disrupted the DGKα/FAK signalosome to inhibit FAK-controlled signaling pathways and the malignant progression of ESCC cells both in vitro and in vivo, whereas produced no toxicity to the normal cells. Molecular validation specifically demonstrated that Asp435 site in the catalytic domain of DGKα contributed to chrysin-mediated inhibition of the assembly of DGKα/FAK complex. This study has illustrated DGKα/FAK complex as a target of chrysin for the first time, and provided a direction for the development of natural products-derived PPIs inhibitors in tumor treatment.

Chrysin inhibited epithelial-mesenchymal transition of type Ⅱ alveolar epithelial cell by regulating NF-κB/Twist 1 signaling pathway / 中国中药杂志

This study aimed to assess whether chrysin(ChR) can inhibit epithelial-mesenchymal transition(EMT) of type Ⅱ alveolar epithelial cell and produce anti-pulmonary fibrosis effect by regulating the NF-κB/Twist 1 signaling pathway. Sixty rats were randomly divided into the control group, the bleomycin(BLC) group, BLC+ChR(50 mg·kg~(-1)) group and BLC+ChR(100 mg·kg~(-1)) group, with 15 rats in each group. The pulmonary fibrosis model was induced by intratracheal injection of BLC(7 500 U·kg~(-1)). Rats were orally administered with different doses of ChR after BLC injection for 28 days. The cells were divided into control group, TGF-β1 group(5 ng·mL~(-1)), and TGF-β1+ChR(1, 10, 100 μmol·L~(-1)) groups. The type Ⅱ alveolar epithelial cells were treated with TGF-β1 for 24 h, and then treated with TGF-β1 for 48 h in the presence or absence of different doses of ChR(1, 10 and 100 μmol·L~(-1)). The morphological changes and collagen deposition in lung tissues were analyzed by HE staining, Masson staining and immunohistochemistry. The mRNA and protein expression levels of collagen Ⅰ, E-cadherin, zonula occludens-1(ZO-1), vimentin, alpha smooth muscle actin(α-SMA), inhibitor of nuclear factor kappa B alpha(IκBα), nuclear factor-kappa B p65(NF-κB p65), phospho-NF-κB p65(p-p65) and Twist 1 in lung tissues and cells were detected by qPCR and Western blot, respectively. The animal experiment results showed that as compared with the BLC group, after administration of ChR for 28 days, bleomycin-induced pulmonary fibrosis in rats was significantly relieved, collagen Ⅰ expression in lung tissues was significantly reduced(P<0.05 or P<0.01), and EMT of alveolar epithelial cells was obviously inhibited [the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], concomitantly with significantly reduced IκBα and p65 phosphorylation level in cytoplasm and decreased NF-κB p65 and Twist 1 expression in nucleus(P<0.05 or P<0.01). The cell experiment results showed that different doses of ChR(1, 10 and 100 μmol·L~(-1)) significantly reduced TGF-β1-induced collagen Ⅰ expression(P<0.05 or P<0.01), significantly inhibited EMT of type Ⅱ alveolar epithelial cells[the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], and inhibited IκBα and p65 phosphorylation in cytoplasm and down-regulated NF-κB p65 and Twist 1 expression in nucleus induced by TGF-β1(P<0.05 or P<0.01). The results suggest that ChR can reverse EMT of type Ⅱ alveolar epithelial cell and alleviate pulmonary fibrosis in rats, and its mechanism may be associated with reducing IκBα phosphorylation and inhibiting NF-κB p65 phosphorylation and nuclear transfer, thus down-regulating Twist 1 expression.

Chemical constituents from Alpinia oxyphylla / 中草药

Objective: To study the chemical constituents of Alpinia oxyphylla. Methods: The ethyl acetate fraction of 95% ethanol extract from A. oxphylla was isolated and purified by silica, MCI, Sephadex LH-20 and semi-preparative HPLC, then the structures of obtained compounds were identified by physicochemical properties and spectral data. Results: Sixteen compounds were isolated and their chemical structures were identified as phthalic acid-bis (2’-ethyl heptanyl) easter (1), (E)-1-(4’-hydroxy-3’-methoxyphenyl)-7- (4″-hydroxy-phenyl)-hept-4-en-3-one (2), 5-hydroxy-7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-3-heptanone (3), 1β,4β,7β- trihydroxyeudesmane (4), bullatantriol (5), 1,5-epoxy-3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxy- phenyl) heptanes (6), 1-tetratriacontanol (7), dihydrogingerenone B (8), tectochrysin (9), chrysin (10), oxyphyllenone B (11), (1R,4R,10R)-1β,4α-dihydroxy-11,12,13-trinor-5,6-eudesmen-7-one (12), daucosterol (13), 1-(4’-hydroxyphenyl)-7-(3″-methoxy- 4″-hydroxyphenyl)-4-ene-3-heptanone (14), yakuchinone A (15) and 5-dehydroxy-hexahydro-demethoxycurcumin B (16). Conclusion: A total of 16 compounds were isolated and identified. The compounds 1-2, 4-8 are isolated from the genus for the first time, and compounds 3 were isolated from the plant for the first time.

UHPLC-MS-MS high throughput analysis-based screening and evaluation of PXR/CYP3A4-induced lipid-regulating quality marker in propolis / 中草药

Objective: To screen and evaluate PXR/CYP3A4-induced lipid-regulating quality marker in propolis with precise and quantitative method. Methods: The LS174T cell was given certain amount of midazolam injection, along with different dosage of known components found in propolis, after incubation and extraction, the samples were determined for 1’-OH-midazolam, and each compound was evaluated to discover the PXR/CYP3A4 pathway regulatory activity according to the results; Then, compounds selected were used as indexes for UHPLC-MS-MS content determination, and their own values were regarded as a preliminary step of confirming PXR/CYP3A4-induced lipid-regulating quality markers of propolis. Results: In all components tested, chrysin, galangin, heterochlorogenic acid A, quercetin, and caffeic acid phenethylester significantly affected the 1’-OH-midazolam yield compared with blank and positive control, indicating their obvious influence on PXR/CYP3A4 expression; The UHPLC-MS-MS determination showed that except galangin, heterochlorogenic acid A, and quercetin, all the other compounds had adequate content in propolis to take effect. Conclusion: Chrysin, galangin, caffeic acid phenethylester, and quercetin were probably defined as PXR/CYP3A4-induced lipid-regulating quality marker in propolis, which inhibited the expression of such targets to down-regulate blood lipid level; Additionally, the method used for quality marker screening and evaluation in this study was fast, effective and quantitative, and capable of carrying out high throughput active component screening for PXR/CYP3A4 regulatory activities.

Mechanism of Tanreqing Capsule on treatment of coronavirus disease 2019 based on network pharmacology / 中草药

Objective: To explore the network regulation mechanism of Tanreqing Capsule (TRQC) on the treatment of coronavirus disease 2019 (COVID-19). Methods: Potential targets of the 19 major constituents in TRQC were predicted by the Swiss Target Prediction server and TCMSP database. Gene ontology (GO) function enrichment and pathway analysis of the targets were analyzed by Omicsbean analytic system and String 10 database. Finally, Cytoscape 3.6.1 software was used to construct the network pharmacology map. Results: A total of 19 compounds affected 68 pathways such as IL-17 signaling pathway, T cell receptor signaling pathway, arachidonic acid metabolism, cAMP signaling pathway, PI3K-Akt signaling pathway, influenza A, etc, by acting on 163 related targets, which associated with anti-inflammation, immune regulation, antipyresis, eliminating phlegm, relieving cough and asthma, analgesia, antibacterial and antiviral, and sedation. The network of "compound-target-pathway-pharmacological action-efficacy" was also constructed. Conclusion: The major constituents in TRQC, including scutellarin, baicalin, oroxylin-7-O-glucuronide, chrysin-7-O-glucuronide, forsythin, forsythiaside E, forsythiaside D, chlorogenic acid, caffeic acid, isochlorogenic acid A, ursodeoxycholic acid and chenodeoxycholic acid, may interfere with efficacy-related biological processes associated with antipyresis, eliminating phlegm and removing toxin by acting on key protein like TNF, EGFR, NOS3, PTGS2, IL2, GABBR1, MAPK14, ADRB2, REN, VCAM1, ACHE, PTPRC, etc.

Overall quality control of Jinzhen Oral Liquid based on HPLC-UVD-ELSD fingerprint and simultaneous determination of 13 main representative components / 中草药

Objective: To establish the fingerprint of Jinzhen Oral Liquid (JOL) by HPLC-UVD-ELSD and determine the main 13 representative components (gallic acid, liquiritin, aloe-emodin-8-O-β-D-glucopyranoside, liquiritigenin, baicalin, chrysin-7-O-β-D- glucoronide, oroxyloside, wogonoside, chrysophal-1-O-β-D-glucopyranoside, chrysophal-8-O-β-D-glucopyranoside, rhein, glycyrrhizic acid, hyodeoxycholic acid and cholic acid) simultaneously, in order to provide reference for the overall quality control of JOL. Methods: The separation was developed on Cosmosil-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution with methanol-water [containing 0.1% formic acid] at 254 nm, the temperature of drift tube was maintained at 115 ℃ and the carrier gas flow rate was 2.0 L/min. An HPLC-UVD-ELSD fingerprint of JOL was set up, and 15 batches of JOL were evaluated by similarity assay. Furthermore, the contents of the main 13 representative components were determined on the premise of small disparities among batches. Results: The HPLC-UVD-ELSD fingerprint of JOL was established with good separation, and 13 chemical components were determined simultaneously. Fifteen main characteristic peaks [gallic acid (peak 1), liquiritin (peak 5), aloe-emodin-8-O-β-D-glucopyranoside (peak 7), liquiritigenin (peak 11), baicalin (peak 13), chrysin-7-O-β-D-glucoronide (peak 16), oroxyloside (peak 17), wogonoside (peak 18), chrysophal-8-O-β-D-glucopyranoside (peak 19), chrysophal-1-O-β-D-glucopyranoside (peak 20), rhein (peak 24), glycyrrhizic acid (peak 26), (18β,20α)-glycyrrhizic acid (peak 27), hyodeoxycholic acid (peak 28), cholic acid (peak 29)] from four formula of JOL were chemically identified and 29 main characteristic peaks were assigned to individual herbs (peaks 8, 12, 13, 15-18 originate from Scutellariae Radix, peaks 3-5, 10, 11, 25-27 originate from Glycyrrhizae Radix et Rhizoma, peaks 1, 6, 7, 9, 14, 19, 20, 24 originate from Rhei Radix et Rhizoma, peak 2 originates from Fritillariae Ussuriensis Bulbus, peaks 28, 29 originate from Bovis Calculus Artifactus, peaks 21-23 originate from auxiliary materials). The similarity of 15 batches of JOL was about 0.968 to 1.000. Moreover, good linear relationships were found (R2=0.999 0-0.999 9), and the average recovery rates were 96.90%-102.84%. The content range of quantitative components in 15 batches of JOL (gallic acid 51.82-148.27 μg/mL, liquiritin 75.04-130.00 μg/mL, aloe-emodin-8-O-β-D-glucopyranoside 31.72-39.84 μg/mL, liquiritigenin 14.24-43.65 μg/mL, baicalin 610.37-867.40 μg/mL, chrysin-7-O-β-D-glucoronide 12.87-34.09 μg/mL, oroxyloside 62.45-101.48 μg/mL, wogonoside 155.41-205.86 μg/mL, chrysophal-1-O-β-D-glucopyranoside 11.56-23.72 μg/mL, chrysophal-8-O-β-D- glucopyranoside 16.14-36.87 μg/mL, glycyrrhizic acid 222.97-310.32 μg/mL, hyodeoxycholic acid 177.48-239.70 μg/mL, cholic acid 98.54-132.85 μg/mL) was determinated. Conclusion: The qualitative and quantitative methods of HPLC-UVD-ELSD mentioned above provided important evidence for further improving the overall quality standard of JOL.

Chemical constituents from stems and leaves of semi-mangrove plant, Barringtonia racemosa / 中草药

Objective: To study the chemical constituents of the stems and leaves of semi-mangrove plant, Barringtonia racemosa. Methods: The chemical constituents of B. racemosa were separated and purified by silica gel, ODS, Sephadex LH-20 gel column chromatographies and preparative HPLC. Their structures were identified by physicochemical properties, spectroscopic analysis, as well as comparisons with the data reported in literature. Results: A total of 17 compounds were isolated from the 90% ethanol extract of the stems and leaves of B. racemosa, which were identified as chrysin (1), ayanin (2), genkwanin (3), rhamnocitrin (4), tricin (5), dillenetin (6), 5,3'-dihydroxy-7,4'-dimethoxyflavone (7), 5,7,3',4'-tetramethoxyflavone (8), 5-hydroxy-6,7,8,3',4'- pentamethoxy flavone (9), petasitolone (10), sarmentol F (11), dehydrovomifoliol (12), blumenol A (13), 10-hydroxyaristolan-9-one (14), alphitolic acid (15), oleanolic lactone (16), and 11,12-dehydroursolic acid lactone (17). Conclusion: All compounds are isolated from the genus Barringtonia for the first time.

Chemical constituents from whole plants of Scutellaria viscidula / 中草药

Objective: To investigate the chemical constituents from the whole plants of Scutellaria viscidula. Methods: By means of preparative HPTLC and column chromatography over silica gel and Sephadex LH-20, the chemical constituents were isolated and purified. Their structures were elucidated by physico-chemical properties and spectral analyses. Results: A total of 22 compounds were isolated and identified as: 5-hydroxy-7,8-dimethoxyflavone (1), chrysin (2), 5,7-dimethoxyflavone (3), 5,7,4’-trihydroxy- 6-methoxyflavone (4), 5,7,4’-trihydroxyflavonoid (5), 5,7,2’-trihydroxy-8-methoxyflavone (6), 5,7-dihydroxy-8,2’-dimethoxyflavone (7), 5,7,2’-trihydroxy-6-methoxyflavanone (8), 5-hydroxy-6,7,8-trimethoxyflavone (9), 5,6,7-trimethoxyflavone (10), 3,5-dimethoxy- 6,7-methylenedioxyflavone (11), 5,2’-dihydroxy-7,8,6’-trimethoxyflavone (12), 5,2’-dihydroxy-7,8,6’-trimethoxyflavanone (13), 5,7-dimethoxyflavanone (14), 7-methoxy-chrysin (15), 2’-hydroxy-5,7,8-trimethoxyflavone (16), ferulic acid (17), 4-hydroxy-3- methoxybenzoic acid (18), p-hydroxy-benzaldehyde (19), 3,5-dimethoxy-4-hydroxybenzoic acid (20), 7-hydroxy-6-methoxy-coumarin (21), 6,7-dihydroxyl-coumarin (22). Conclusion: Compounds 1, 3-16, 18 and 20 are isolated from Scutellaria viscidula for the first time.

In vitro antimicrobial and antimycobacterial activity and HPLC-DAD screening of phenolics from Chenopodium ambrosioides L

Abstract The main objective of this study was to demonstrate the antimicrobial potential of the crude extract and fractions of Chenopodium ambrosioides L., popularly known as Santa-Maria herb, against microorganisms of clinical interest by the microdilution technique, and also to show the chromatographic profile of the phenolic compounds in the species. The Phytochemical screening revealed the presence of cardiotonic, anthraquinone, alkaloids, tannins and flavonoids. The analysis by HPLC-DAD revealed the presence of rutin in the crude extract (12.5 ± 0.20 mg/g), ethyl acetate (16.5 ± 0.37 mg/g) and n-butanol (8.85 ± 0.11 mg/g), whereas quercetin and chrysin were quantified in chloroform fraction (1.95 ± 0.04 and 1.04 ± 0.01 mg/g), respectively. The most promising results were obtained with the ethyl acetate fraction, which inhibited a greater number of microorganisms and presented the lowest values of MIC against Staphylococcus aureus and Enterococcus faecalis (MIC = 0.42 mg/mL), Pseudomonas aeruginosa (MIC = 34.37 mg/mL), Paenibacillus apiarus (MIC = 4.29 mg/mL) and Paenibacillus thiaminolyticus (MIC = 4.29 mg/mL). Considering mycobacterial inhibition, the best results were obtained by chloroform fraction against M. tuberculosis, M. smegmatis, and M. avium (MIC ranging from 156.25 to 625 µg/mL). This study proves, in part, that the popular use of C. ambrosioides L. can be an effective and sustainable alternative for the prevention and treatment of diseases caused by various infectious agents.

Chrysin promotes SMMC-7721 cell apoptosis by regulating MAPKs signaling pathway / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of chrysin in inducing apoptosis of human hepatic carcinoma cells and explore the possible mechanism.</p><p><b>METHODS</b>Human hepatic carcinoma SMMC-7721 cells treated with DMSO or chrysin at different concentrations (5-200 μg/mL) were examined for changes in the cell proliferation using CCK-8 assay. The morphological changes of SMMC-7721 cells were observed in response to treatment with 5, 10, or 20 μg/mL chrysin, and the changes in the cell nuclei were observed using DAPI nuclear staining. Annexin Ⅴ-FITC/PI flow cytometry was used to determine the cell apoptosis rate. The changes in the apoptosis-related proteins (PARP and caspase-3) and MAPKs signal pathway were detected with Western blotting.</p><p><b>RESULTS</b>Chrysin treatment obviously suppressed the proliferation of SMMC-7721 cells in a dose-dependent manner below the concentration of 60 μg/mL. Chrysin (20 μg/mL) also caused significantly increased cell apoptosis and significant cleavage of PARP and caspase-3. Chrysin significantly activated MAPKs signaling pathway in a time-and dose-dependent manner, with the peak activation level occurring at 15 min. Pretreatment of the cells with specific inhibitors of the MAPKs pathway obviously inhibited the effect of chrysin in inducing cell apoptosis.</p><p><b>CONCLUSIONS</b>Chrysin inhibits the proliferation and promotes apoptosis of SMMC-7721 cells by regulating the activation of MAPKs signaling.</p>

Mechanism of interaction between chrysin and leptin / 中国中药杂志

The study of interaction mechanism between chrysin and leptin was investigated by various spectroscopic techniques and atom force microscope. The ultraviolet spectrum presents a red shift in 200-220 nm after chrysin upon. And there is a structure alternative showed in 270 nm when the concentration ratio of chrysin and leptin in 10-15. From the fluorescence spectrum, it was found that chrysin could combine with leptin in physiological condition. The binding constant (Ka) values, at 298 K and 310 K, were (0.41±0.05)×10⁶ and (3.26±0.46)×10⁶ L·mol⁻¹, and the binding site number were 1.02±0.04 and 0.51±0.01, respectively. The atom force microscope results showed that the dimension of leptin molecules became more swollen after binding with chrysin because of the hydrophobicity. These results demonstrate that the mechanism of chrysin and leptin interaction could play a role in leptin adjust in human body, and it could provide a new aspect for the study of obesity treatment.

Synthesis and anti-tumor activities of novel 7-O-amino acids chrysin derivatives / 中草药·英文版

Objective: To design and synthesize a series of chrysin derivatives and evaluate the antitumor activities with MTT assay, so as to investigate molecular structure-activity relationship with molecular docking. Methods: Target products were synthesized with high yield by substitution reaction, hydrolysis reaction, esterification reaction, and saponification reaction in sequence, and activities of all compounds were evaluated with human gastric carcinoma cell lines MGC-803 and human breast carcinoma cell lines MCF-7 through standard MTT assay. Molecular docking results were calculated with Surflex Geom X programme of Sybyl X-2.0 version workstation. Results: 7-O-amino acids chrysin derivatives 6a–6l were synthesized and their inhibitory effects were evaluated by comparing the material chrysin with positive control drug 5-fluorouracil (5-FU). Among these derivatives, compound 5b (IC50 = 24.50 ± 2.26 µmol/L), 5k (IC50 = 24.30 ± 2.19 µmol/L), and 6f (IC50 = 24.61 ± 2.01 µmol/L) showed better inhibitory activities against MGC-803 cell lines, and compound 5g (IC50 = 13.15 ± 1.73 µmol/L) and 5j (IC50 = 12.34 ± 1.25 µmol/L) showed better inhibitory activities against MCF-7 cell lines than chrysin and 5-FU. Molecular docking scores showed a credible consistency compared with MTT results. Conclusion: Compounds 5b, 5d, 5g, 5j, 5k, and 6f showed good antiproliferative effects on specific tumor cells, and compound 5g should be researched further when according to molecular docking.

Comparative analysis of species and contents of flavonoids before and after honey refining / 中草药

Objective To study the changes of flavonoid components in honey before and after refining. Methods Six batches of honey from different sources were collected and refined based on Chinese Drugs Pharmacy. Solid phase extraction was used to enrich flavonoids from samples. HPLC-TQ-MS was established to determine the contents of 15 flavonoids in crude and refined honey, the separation was performed on a Thermo Accucore RP-MS (100 mm × 2.1 mm, 2.6 μm) column with the gradient elution of methanol-0.1% formic acid water, the flow rate was 0.3 mL/min, and the column temperature was 30 ℃. MS condition: Electrospray ionization (ESI) source was applied and operated in the positive multiple reaction monitoring (MRM) modes. Results Twelve kinds of flavonoids in three species, flavonoids, dihydroflavonoids, and isoflavones, were detected in honey, which including quercetin, morin, xanthophyllin, kaempferol, apigenin, wogonin, galangin, chrysin in flavonoids; pinobanksin, naringin, and pinocembrin in dihydroflavonoids; genistein in isoflavones. There was significant difference in the species and contents of flavonoids between crude honey and refined honey. After refining, rutin and narirutin were detected which have not been reported in references, and the contents of 12 kinds of flavonoids have increased at different degrees. The contents of quercetin, morin, xanthophyllin, kaempferol, apigenin, wogonin, pinocembrin, and chrysin in linden honey have increased, but the contents of pinobanksin, naringin, and galangin have decreased; all the contents of the components in acacia honey have increased except apigenin, especially the quercetin and morin; The content of 12 kinds of flavonoids in the honey of various flowers and Chinese date honey all have increased. Conclusion Refining can change the species and contents of flavonoids in honey.

Preparation of chrysin solid lipid nanoparticles and their pharmacokinetic behaviors / 中成药

AIM To prepare chrysin solid lipid nanoparticles and to evaluate their pharmacokinetic behaviors.METHODS The particle size,Zeta potential and in vitro release rate of nanoparticles prepared by emulsification uhrasonication-low temperature solidification method were determined.Twelve SD rats were randomly divided into two groups and were intragastrically given suspensions of crude drug and nanoparticles,respectively.HPLC was used for the content determination of chrysin in plasma,after which blood drug concentration-time curves were drawn,and pharmacokinetic parameters were calculated.RESULTS The obtained nanoparticles demonstrated the particle size of (207.15 ±30.59) nm,PDI of (0.224 ±0.067) and Zeta potential of (-34.8 ±5.9) mV,respectively,whose accumulative release rate reached 84.36％ within 36 h.Their Cmax [(9.04 ± 1.52) μg/mL] and AUC0～t,[(33.67 ± 3.47) μg · h/mL] were much higher than those of the crude drug (P ＜ 0.01).CONCLUSION Solid lipid nanoparticles can promote the oral absorption and bioavailability of chrysin,with significant sustained-release characteristics.

Effects of chrysin on impaired vascularendothelial function induced by high glucose / 中国药理学通报

Aim To explore the effects of chrysin on endothelial dysfunction induced by acute high glucose.Methods ① The effects of chrysin on normal isolated aortic at contraction induced by PE and on endothelial dysfunction induced by high glucose were tested in the following medium: normal group,chrysin group;normal-glucose group: glucose 11mmol·L-1 in Krebs' solution;high-glucose group: glucose 44 mmol·L-1 in Krebs' solution;mannitol group: mannitol 33 mmol·L-1 in Krebs' solution and chrysin group: 44 mmol·L-1 Glu+chrysin 1.0 μmol·L-1 in Krebs' solution.② The effects of chrysin on HUVEC cell viability after incubated in high glucose were observed in the following groups: normal-glucose group: glucose 5.5 mmol·L-1 in culture solution;high-glucose group: glucose 33.3 mmol·L-1 in culture solution;mannitol group: mannitol 27.8 mmol·L-1 in culture solution and chrysin group: chrysin(25,50 μmol·L-1)in culture solution.And the NO release was also testd in these groups.Results ① Chrysin could induce vaso-dilation in a dose-dependent manner at normal glucose.The Emax was(58.94±9.61)％,and the EC50 value was 51.9 μmol·L-1.After incubating the aortic rings with high glucose(44 mmol·L-1)for 4 h,there were significant differences in ACh-induced vascular relaxation between the normal glucose group and the high glucose group.The Emax was(32.12±3.92)％and the EC50 value was 78.0 μmol·L-1 of high glucose group(P<0.01).The endothelium-independent relaxation induced by SNP was not significantly different between the two groups.And chrysin(1.0 μmol·L-1)could reverse the decline of ACh-induced vasorelaxation response induced by high glucose(44 mmol·L-1).The Emax was(70.7±3.87)％and the EC50 value was 0.852 μmol·L-1.② The cell viability of HUVEC was depressed after incubated in high glucose,and chrysin could reverse the decline in a concentration-dependent way.And chrysin in defferent concentrations could increase the cell NO release.Conclusion Chrysin could prevent the acute high glucose-induced vascular endothelial dysfunction and could increase the NO release.