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Objective:To explore the potential molecular mechanism of corylin in the treatment of lung cancer. Method:A549 cells were treated with different concentrations of corylin， and their proliferation was detected using methye thiazolye telrazlium （MTT） reagent. Then the trend analysis and gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were conducted to screen the key genes and pathways of corylin against A549 cell proliferation， followed by the verification of sequencing results by real-time polymerase chain reaction （Real-time PCR）. Result:Corylin inhibited the proliferation of A549 cells and regulated the expression of 4 364 genes in cells. The trend analysis revealed that these genes were clustered into 20 distinct modules， among which four were significantly down-regulated， suggesting that corylin exerted the anti-proliferation effect by inhibiting the expression of some genes. The inter-group comparison of differentially expressed genes （DEGs） showed that the elevation in the concentration of corylin resulted in more down-regulated genes but weakened proliferation， consistent with the findings by trend analysis. The GO and KEGG enrichment analysis of 278 DEGs in the high-dose corylin group demonstrated that corylin mainly changed the cellular and metabolic processes， which was attributed to its regulation of steroid biosynthesis， fatty acid metabolism， biosynthesis of unsaturated fatty acids， synthesis and degradation of ketone bodies， and steroid hormone biosynthesis. The Real-time PCR results confirmed that corylin down-regulated the mRNA expression levels of LSS， stearoyl-CoA desaturase （SCD）， 3-hydroxy-3-methylglutaryl-CoA synthase 1 （HMGCS1）， but up-regulated the mRNA expression of recombinant human angiopoietin-like protein 4 （ANGPTL4）， basically consistent with the transcriptomics results. Conclusion:Corylin inhibits A549 cell proliferation and alleviates lung cancer by targeting the related genes in lipid metabolism pathways.
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BACKGROUND: The balance of bone homeostasis is mediated by the osteoclast-related bone resorption and osteoblast-related bone formation. Over-activation of osteoclasts results in a series of bone metabolic diseases including rheumatoid arthritis and osteoporosis. The activation of nuclear factor-κB pathway induced by receptor activator of nuclear factor-κB ligand (RANKL) plays an important role in osteoclastogenesis. OBJECTIVE: To explore the effect of corylin on RANKL-mediated osteoclastogenesis. METHODS: RAW264.7 cells were incubated with 0, 1, 2, 4, 8, 16, 32, 64, 128 μmol/L corylin. The cytotoxicity of corylin was detected by cell counting kit-8 assay. RANKL induced the differentiation of RAW264.7 cells into osteoclasts, during which 2, 5, 10 μmol/L corylin was given. The number of osteoclasts was analyzed by TRAP staining after 5 days of intervention and the morphology and function of osteoclasts were analyzed by F-actin staining. Bone resorption assay was conducted after 2 days of intervention. The activation of nuclear factor-κB pathway was detected by western blot at 0, 15, 30, and 60 minutes of intervention. Then in vivo experiments were carried out, and the ovariectomized mice were intraperitoneally given 10 mg/kg twice a week. After 6 weeks of intervention, mouse femurs were taken for morphological analysis. RESULTS AND CONCLUSION: There was no cytotoxicity of corylin below the concentration of 16 μmol/L. Corylin inhibited osteoclastogenesis in a dose-dependent manner. Corylin inhibited the formation of F-actin and resorption activity of osteoclasts. Corylin inhibited RANKL-mediated nuclear factor-κB pathway. Corylin treatment reduced the bone loss in postmenopausal osteoporosis mice. Overall, corylin inhibits osteoclastogenesis via blocking nuclear factor-κB pathway and attenuates postmenopausal osteoporosis.
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Aim: To investigate the regulation effect and molecular mechanism of corylin on NLRP3, NL-RC4, AIM2 inflammasomes with immortalized bone marrow-derived macrophages. Methods: The effect of corylin on the NLRP3, NLRC4, AIM2 inflammasome activation was evaluated with LPS-induced ATP, nigericin, salmonella, poly(dA: dT). Caspase-1 activity was determined by Caspase-Glo® 1 Inflammasome Assay. Western blot was performed to observe the protein expression levels of mature IL-1β, caspase-1 p20 in the culture supernatants, pro-caspase-1, pro-IL-1β, ASC, NLRP3 in the cell lysates. Results: Corylin blocked the self-slicing of pro-caspase-1 induced by ATP, nigericin, salmonella and poly(dA: dT), then suppressed the secretion of mature IL-1β mediated by caspase-1, which showed that corylin inhibited the NL-RP3, NLRC4, AIM2 inflammasomes activation. Moreover, corylin irreversibly attenuated the activation of NLRP3 inflammasome without affecting NF-κB signaling pathway. Conclusions: Corylin inhibits the inflammasome activation of NLRP3, NLRC4, and AIM2, and further reduces its mediated immune inflammatory response. Meanwhile, it provides new ideas and strategies for the treatment of immune inflammatory diseases by using corylin-related preparations.
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Objective: To establish UPLC method for the simultaneous determination of 10 components, such as psoralen, isopsoralen, psoralidin, bavachinin, isobavaenin, corylifolin, isobavachalcone, corylin, neobavaisoflavone, and bakuchiol in Psoralea Fructus, and to study the effect of processing time on 10 components in stir-frying Psoralea Fructus with salt solution. Methods: The chromatographic separation was achieved on a C18 column (50 mm×4.6 mm, 1.8 μm) with acetonitrile (A)-water (B) as mobile phase at the flow rate of 1.0 mL/min for gradient elution; The column temperature was 30℃. The determination wavelength was 250 nm. Results: The 10 components were well separated within 15 min. The RSD values of reproducibility were less than 3%. The stability was good in 24 h. The linear relationship between the concentration and peak areas of the 10 components was good (r≥0.9970). The average recoveries were 96%-105% and the RSD values were all less than 3%. Conclusion: The method is simple, reliable and accurate, could be used for the quality control of Psoralea Fructus. With processing time prolonged, the content of coumarins showed first decreased, then increased and last decreased. Flavonoids, bakuchiol and the total content of 10 components above were decreased.
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Objective: To optimize the processing technology for Psoraleae Fructus by D-optimal response surface methodology with UHPLC. Methods: Based on the content variations of nine main constituents (psoralen, isopsoralen, neobavaisoflavone, bavachin, psoralidin, corylifolinin, corylin, bavachalcone, and bakuchiol) determined by UHPLC, the D-optimal response surface methodology combined with single factor experiment was used to optimize the moistened time, stir-fried temperature, salted dosage, and stir-fried time in the processing technology for Psoraleae Fructus. Results: The optimal processing technology for Psoraleae Fructus was as follows: 100 g Psoraleae Fructus was mixed with 2.10 g salt, moistened for 12 h, and stir-fried for 30 min at 80℃. Conclusion: The processing technology for Psoraleae Fructus optimized by D-optimal response surface methodology with UHPLC could be used to optimize the process parameters, to promote the stability and repeatability of the processing technology, which provides the technique support for improving the quality uniformity of Psoraleae Fructus and safety in clinic application.
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Objective: To study the chemical constituents of Cremastra appendiculata and their neuroprotection. Methods: The chemical constituents were separated and purified by silica gel, Sephadex LH-20 column chromatography, ODS column chromatography, and HPLC column chromatography. Their structures were determined by NMR spectrum. Results: Applying various chromatographic separation methods, 23 compounds were obtained from the aceticether extraction of 55% ethanol extract of C. appendiculata. Based on physicochemical properties and spectroscopic analysis, 20 structures were identified as miltarine (1), 1-(4-β-D-glucopyranosyloxybenzyl)-2-isobutylmalate-4-methyl (2), 1-[4-(β-D-glucopyranosyloxy)benzyl]-4-methyl-2-benzylmalate (3), batatasin III-3-O-glucoside (4), eucomic acid (5), hesperidin (6), (4-methyl-1,3-phenylene) dicarbamic acid methyl ester (obtucarbamate A) (7), (3-meth-oxycarbonylamino-2-methyl-phenyl)-cabamic acid metylester (8), 4,4'-diphenylmethanebis(methyl) carbamate (9), 3,3'-dihydroxy-2',6'-bis(p-hydroxybenzyl)-5-methoxybibenzyl (10), 3',5-dihydroxy-2-(4-hydroxybenzyl)-3-methoxybibenzyl (11), 3,3'-dihydroxy-2-(4-hydroxybenzyl)-5-methoxybibenzyl (12), corylin (13), batatasin III (14), neobavaisoflavone (15), isobavachalcone (16), 2,6,2',6'-tetramethoxy-4,4-bis(2,3-epoxy-1-hydroxypropyl)biphenyl (17), coelonin (18), gigantol (19), and β-sitosterol (20). Conclusion: Compounds 7-9, 13, and 15-17 are obtained from the plants of Cremastra Lindl. for the first time. Compounds 5 and 6 are obtained from the plant for the first time. Compounds 1 and 17 have the good performance of neuroprotection in hydrogen peroxide model. Neuroprotective mechanism is likely to be achieved by weakening the oxidative damage.