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Background: Silver Nanoparticles are drawing significant attention from the scientific community to explore a wide range of its medical applications. Human body is under constant stress due to free radicals generated by the physiological and pathological conditions in the body. Scavenging systems or Antioxidants can help alleviate the damages caused by these radicals which can influence the course of progress in several chronic diseases with an inflammatory background. External antioxidants supplement and facilitate the overwhelmed scavenging systems in the body.Silver Nanoparticles can enhance the therapeutic effects of phytochemicals. Aim: To Synthesize silver nanoparticles using the phytochemical Hesperidin and studying its Free radical scavenging activity. Methods: Silver Nanoparticles are synthesized using chemical reduction method. The synthesis is confirmed using spectrophotometric studies. Free Radical scavenging activity is detected using 1, 1-diphenyl-2-picrylhydrazyl (DPPH �) free radical scavenging assay. Results: Silver nanoparticles were successfully synthesized which was confirmed by the change in color of the solution and peak absorbance peak at 420 nM on spectrophotometric studies.Hesperidin Silver Nanoparticles exhibited higher free radical scavenging activity when compared with pure hesperidin and standard Ascorbic acid. Conclusion: Hesperidin can ideally be used for the synthesis of silver nanoparticles and the synthesized Silver Nanoparticles enhances the free radical scavenging activity of Hesperidin which can further be evaluated by In Vivo studies.
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Background: Silver Nanoparticles are extensively studied by the scientific community for therapeutic applications. With respect to the fundamental pillars of bioethics 揚rimum non nocere� equal emphasis should be given to evaluate the toxicological perspectives of Silver nanoparticles. This study aims at evaluating the InVitro cytotoxic effects of Silver nanoparticles synthesized using hesperidin. Aim: To study the In Vitro cytotoxicity of silver nanoparticles on PBMC cells using (3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Methods: Synthesized silver nanoparticles at various concentrations are incubated with peripheral blood mononuclear cells (PBMC). After 24 hours MTT is added to the mixture to evaluate the cell viability post incubation. Yellow MTT (a tetrazole) which is reduced to purple formazan in the mitochondria of living cells. The absorbance of this colored solution can be quantified by measuring at 570 nm by a spectrophotometer. This reduction takes place only when mitochondrial reductase enzymes are active, and therefore conversion can be directly related to the number of viable (living) cells. Results: ?.Conclusion: Silver Nanoparticles do not exhibit any significant cytotoxicity on PBMCs and also there were no dose dependent trends in the results.
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Objetivo: elaborar una bebida por fermentación alcohólica y la cuantificación de flavonoides del zumo de Citrus x clementina (naranja). Metodología: se utilizó el método de fermentación alcohólica por levadura de la variedad Saccharomyces cerevisiae, se fermento el jugo de naranja con una densidad de 1,050 glcm3 por 5 semanas y se cuantificó los flavonoides de la bebida alcohólica por el método de cromatografía HPLC. Resultados: después de las 5 semanas se analizó que la bebida por fermentación alcohólica tuvo un 11 % de alcohol y flavonoides de hesperidina 13,9 mgl100 ml y naringenina 6,3 mg/100 ml en su concentración.
SUMMARY Aim: to elaborate a drink by alcoholic fermentation and the quantification of flavonoids in Citrus x clementine (orange) juice. Methodology: the method of alcoholic fermentation by yeast of the Saccharomyces cerevisiae variety was used, the orange juice was fermented with a density of 1.050 glcm3 for 5 weeks and the flavonoids of the alcoholic beverage were quantified by the HPLC chromatography method. Results: after 5 weeks it was analyzed that the drink by alcoholic fermentation had 11 % alcohol and hesperidin flavonoids 13.9 mgl100 ml and 6.3 mg/100 ml naringenin in its concentration.
Objetivo: elaborar uma bebida por fermentação alcoólica e quantificação de flavonóides no suco Citrus x clementina (laranja). Metodologia: foi utilizado o método de fermentação alcoólica por levedura da variedade Saccharomyces cerevisiae, o suco de laranja foi fermentado com densidade de 1,050 glcm3 por 5 semanas e os flavonóides da bebida alcoólica foram quantificados pelo método de cromatografía HPLC. Resultados: após 5 semanas foi analisado que a bebida por fermentação alcoólica continha álcool a 11 % e flavonóides de hesperidina 13,9 mgl100 ml e 6,3 mg/100 ml naringenina em sua concentração.
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OBJECTIVE@#Post-traumatic stress disorder (PTSD) is a psychiatric disorder characterized by depression and anxiety, that arises due to an imbalance of neurotransmitters in response to excessive stress. Hesperidin (HSD) is a naturally occurring flavonoid shown to exert a variety of biological activities, including antioxidant, anti-inflammatory, and neuroprotective effects.@*METHODS@#This study was used the open field test (OFT) and forced swimming test (FST) to examine the effects of HSD on the depression-like response of rats after exposure to a single prolonged stress (SPS) leading to the dysregulation of the serotonergic activation system. Male rats were given HSD (20, 50, and 100 mg/kg, intraperitoneal injection, n=6-7 per group) once daily for 14 days after exposure to SPS. The influence of administration of HSD on SPS-induced behavioral responses and concentrations of serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), and monoamine oxidase-A (MAO-A) in the rat brain were also investigated using enzyme-linked immunoassays (ELISAs).@*RESULTS@#Daily HSD administration signifificantly improved depression-like behaviors in the FST (P0.05), increased the number of lines crossed in the central zone of the OFT (P0.01), and reduced freezing behavior both in contextual and cued fear conditioning. HSD treatment also attenuated the reduction in SPS-induced 5-HT concentrations in the hippocampus and amygdala. This increase in 5-HT concentrations during HSD treatment was partially attributed to a decrease in the 5-HIAA/5-HT ratio in the hippocampus of rats with PTSD. Furthermore, HSD treatment inhibited activity of MAO-A and decreases of tryptophan hydroxylase-1 expression in the hippocampus.@*CONCLUSION@#HSD was shown to exert antidepressant effects in rats exposed to SPS, suggesting that this natural flflavonoid may be an effective medicine for PTSD.
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Dengue viral infection becomes highly epidemic and rashes the economic stability of most of the developing countriesdue to its wide prevalence with limited therapeutic ailments. Alarming demographic data urge the need for thedevelopment of new antiviral agents which are safe and efficacious. This study aimed to evaluate the antiviral potentialof bioflavonoids (apigenin, hesperidin, kaempferol, myricetin, and naringenin) against dengue virus nonstructural(NS)5 RNA-dependent RNA polymerase (RdRp) by AutoDock and tox prediction tools. The results of moleculardocking analysis strongly suggested that the lead phytocomponents such as apigenin, hesperidin, and kaempferolreveal potential RdRp inhibition as ascertained by its interaction with core active amino acid residues (710 SER, 729ARG, and 737 ARG) on the target. Apigenin exhibited the best binding affinity of −8.28kcal/mol with RdRp, followedby kaempferol (−7.00 kcal/mol), myricetin (−4.37 kcal/mol), naringenin (−4.35 kcal/mol), and hesperidin(−3.20 kcal/mol). The present research finding clearly advocates that plant-derived bioflavonoids possess excellent antiviralproperty against the selected target.
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Objective: To establish the UPLC specific chromatogram and HPLC content determination methods of multi-index components about the material reference of classical Huaganjian and build its quality control system. Methods: According to the ancient books and combining with the previously inspected process, 18 batches of Huaganjian material reference from different origins were prepared. The specific chromatogram was established by using UPLC. Similarity was calculated by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2012). Combining with orthogonal partial least squares discriminant analysis, we excavated the main components that affected the quality of Huaganjian material reference from different batches and origins. Three of these index components (paeoniflorin, hesperidin, paeonol) from prescription sovereign drug, minister drug, and assistant drug were selected and used as indicators for content determination of Huaganjian material reference. HPLC content determination methods were established and the content of 18 batches of samples was determined respectively. Results: The similarity of the specific chromatogram was ≥ 0.989. Thirty-three common peaks were calibrated, and eight common peaks were identified by chemical composition (gallic acid, geniposide, paeoniflorin, hesperidin, didymin, paeonol, sinensetin, and 3,5,6,7,8,3',4'- heptamethoxyflavone). Nine index components that affected the stability between batches were found out (Peak 31, 20, 11, 13, 22, 33, 21, 29, 1). Paeoniflorin, hesperidin, and paeonol were selected as content determination indicators. The content range of these components in material reference was 1.28%-1.95% paeoniflorin, 0.91%-1.02% hesperidin, 0.48%-0.57% paeonol. Conclusion: The quality control method of the material reference of classic prescription Huaganjian was established preliminarily through the UPLC specific chromatogram and HPLC content determination of index components. This method was rapid, simple, feasible, reproducible, stable and could provide a theoretical basis for the subsequent development and quality control of Huaganjian preparations.
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Objective:To establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium and evaluate its quality. Method:According to the preparation conditions of the standard decoction,15 batches of standard decoction of Citri Reticulatae Pericarpium were prepared.HPLC was employed to determine the content of hesperidin in this standard decoction.Ultraviolet spectroscopy(UV) and infrared spectroscopy(IR) were used to establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium.The correlation coefficient method and double index sequence analysis method were used to compare and analyze the spectra of different batches of this standard decoction. Result:The content of hesperidin in 15 batches of this standard decoction were 0.82%-2.60%,and the measured value of dry extract rate was 32.02%-46.11%.Compared with ultraviolet and infrared control fingerprint,the fingerprint similarities of the standard decoction of each batch were > 0.897 and > 0.942,respectively.The double index analysis results showed that the common peak ratio was more than 62.50%,variation peak ratio was less than 46.67%. Conclusion:The quality evaluation method established in this study can be used for systematic evaluation of standard decoction of Citri Reticulatae Pericarpium,and it can provide theoretical reference for the formulation of quality standard of Citri Reticulatae Pericarpium dispensing granules and other related preparations.
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Objective: To establish an high performance liquid chromatography (HPLC) method for the simultaneous determination of four constituents in Niuhuang Qingwei pill (narirutin,naringin,hesperidin,and neohesperidin), and identify the source of Fructus Aurantii Immaturus. Method: The analysis was performed on a Waters CORTECS C18 column (4.6 mm×50 mm,2.7 μm), with acetonitrile-0.12% formic acid solution as the mobile phase for gradient elution. The flow rate was 0.5 mL·min-1, the detection wavelength was set at 283 nm, and the column temperature was 27℃. Result: 12 batches of Niuhuang Qingwei pills showed the different content of flavonoids as Citrus aurantium and C. sinensis. Narirutin,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 5.47-2 735 ng (r=0.999 6),7.25-3 625 ng (r=0.999 5),8.41-4 205 ng (r=0.999 4) and 8.36-4 180 ng (r=0.999 5),and their average recoveries were 101.3% (n=6,RSD 2.9%),98.0% (n=6,RSD 1.8%),95.9% (n=6,RSD 0.8%) and 96.0% (n=6,RSD 1.1%), respectively. The contents of narirutin,naringin,hesperidin,neohesperidin and total flavonoids were 0.36-1.28,2.66-4.87,1.02-11.07,3.58-6.41,and 7.98-13.34 mg·g-1, respectively. Conclusion: The developed method was simple,accurate and reliable,which can be used to identify the source of Aurantii Immaturus Fructus and simultaneously determine the content of four flavonoids in Niuhuang Qingwei pills. It could provide basic research for quality control and composition comparison of 2 kinds of Niuhuang Qingwei pills, showing more comprehensive indicators and reference value for the quality standard improvement of Niuhuang Qingwei pill.
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Objective To establish an HPLC method for the fingerprints analysis of decoction pieces of tangerine peel, so as to provide reference for the quality control of it. Methods Fingerprints of 17 batches of decoction pieces of tangerine peel were established by HPLC and evaluated by cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least square discriminate analysis (OPLS-DA). Results The method of fingerprint of decoction pieces of tangerine peel was established, the similarities were greater than 0.9. There were 25 common peaks in the HPLC fingerprint, of which variable importance projection (VIP) of 14 peaks were greater than 1. Compared with the spectrogram of reference substances, peak 13 was narirutin, peak 14 was hesperidin, peak 23 was nobiletin, peak 24 was 3,5,6,7,8,3’,4’-heptamethoxy flavone, and peak 25 was hesperetin. Conclusion This simple and reliable method can be used for the identification and quality control of decoction pieces of Citri Reticulatae Pericarpium.
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Objective: To prepare a new hesperidin nanoemulsion (HDN-NE) with glycyrrhizic acid as emulsifier, by which could develop a “new green nano-pharmaceutics” of hesperidin. Methods HDN-NE was prepared by high-speed shearing and high-pressure homogenization. The prescription of HDN-NE was optimized with particle size, PDI, and appearance as indexes. The physicochemical property and stability of HDN-NE prepared by the optimal prescription were studied. Results: The optimal prescription of HDN-NE was as follow: The content of hesperidin, glycyrrhizic acid, and oil phase were 0.1%, 0.3%, and 5%, respectively. The shear rate was 13 000 r/min, the cutting time was 2 min, the homogeneous pressure and times were 100 MPa and 6, severally. The result showed that the prepared HDN-NE had the mean size of (262.7 ± 3.1) nm, PDI of 0.234 ± 0.009, Zeta potential of (-35.42 ± 0.72) mV, and solubility of (460.3 ± 2.1) μg/mL. The physicochemical property study showed that the conductivity was (116.4 ± 1.7) μs/cm, the pH was 6.820 ± 0.008, and the turbidity was 451 cm-1 (n = 3). It was identified as O/W emulsion by dyeing method. The droplets were spherical and uniform by transmission electron microscopy. The stability study showed that HDN-NE had good stability. Conclusion: HDN-NE with glycyrrhizic acid as an emulsifier can significantly improve the solubility and stability of hesperidin, which is a new potential nano-drug with safety.
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Objective: To analyze and compare the contents of 12 effective chemical constituents in Aurantii Fructus from different areas, and identify their source of origin by using principal component analysis and discriminant analysis. Methods: Using isonaringin, narigin, hesperidin, neohesperidin, meranzin hydrate, naringin, hesperidin, meranzin, marmin, nobiletin, tangeretin, and auraptene as index, HPLC method was used to determine the content, and the average percentage content was analyzed by principal component analysis and discriminant analysis. Results: Principal component analysis results showed that Aurantii Fructus from different areas had certain similarities, and the content of Aurantii Fructus from Jiangxi Province was relatively stable. Discriminant analysis results showed that the samples from Zhangshu, Yichun, Yuanjiang, Huaihua, Jinhua, and Lanxi could be distinguished, while the samples from Xin’gan in Jiangxi, Jinhua, and Lanxi in Zhejiang overlapped. Conclusion: This method is simple, convenient and reliable, which can be used to screen out the suitable areas for the growth of Aurantii Fructus, so as to lay a foundation for explaining the authenticity of Aurantii Fructus.
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Objective: To optimize the extraction process parameters of seven herbs in Xiaoruzeng Capsules (XC). Methods: The content of paeoniflorin, ferulic acid, hesperidin, imperatorin and ligustilide was determined by HPLC. The extraction rate of five kinds of index components, the dry extract yield of extracted herbs and the fingerprint similarity of extracts were comprehensively evaluated. The orthogonal experiment was carried out to investigate the effects of ethanol concentration, extraction solvent, extraction time and extraction times on the extraction process. The information entropy weighting method was used to determine the objective weight of each index, and the extraction process parameters of seven herbs in XC were optimized. Results: According to the comprehensive scoring results, it was determined that the best extraction process of the preparation was to add six times the amount of 50% ethanol, and decocted two times for 2 h each time. The average score of the three batches of verification scores was 99.72, and the RSD was 0.24%. Conclusion: The preferred process has high extraction rate, with good stability and repeatability, which is suitable for mass production of the preparation.
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Objective: To establish HPLC method for the simultaneous determination of saikosaponin a, naringin, paeoniflorin, calycosin-7-glucoside, tanshinone IIA, cinnamaldehyde, schisandrin, syringin, berberine hydrochloride, chrysophanol and hesperidin in Yigan Yiqi Jieyu Granules (YYJG), and conduct a quality assessment using principal component analysis. Methods: The chromatographic separation was achieved on an Caprisil AQ-C18 (150 mm × 4.6 mm, 5.0 μm) column with mobile phase consisted of 0.1% phosphate-acetonitrile for gradient elution, at the flow rate of 0.8 mL/min; The column temperature was 45 ℃. The results of the content were then combined with the principal component analysis to achieve the scientific assessment of the different batches of drugs. Results: The content of saikosaponina, naringin, paeoniflorin, calycosin-7-glucoside, tanshinone IIA, cinnamaldehyde, schisandrin, syringin, berberine hydrochloride, chrysophanol and hesperidin in YYJG had good linear relationship in the ranges of 1.6-80.0, 14-700, 10-500, 1.6-80.0, 1.6-80.0, 2.4-120.0, 1.2-60.0, 1.2-60.0, 8.0-400.0, 2.0-100.0, and 2.0-100.0 μg/mL, respectively; The average sample recovery rate range were 98.3%, 99.2%, 98.8%, 99.3%, 101.9%, 97.5%, 99.8%, 101.7%, 101.1%, 102.5%, and 100.9% (RSD < 2.0%); The content of 11 active ingredients in 16 batches of samples respectively were 0.233-0.322, 3.007-3.142, 2.201-2.273, 0.320-0.355, 0.317-0.399, 0.451-0.523, 0.265-0.297, 0.209-0.226, 1.848-1.873, 0.380-0.425, and 0.615-0.647 mg/g, respectively. Conclusion: The established method is simple, accurate and reproducible, and can provide the reference for the quality control of YYJG.
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Objective: To investigate the effect of hesperidin on apoptosis of gastric cancer AGS cells and its related molecular mechanisms. Methods: MTT assay was used for the killing effect of hesperidin on human gastric cancer AGS cells; Annexin V-FITC/PI double staining and flow cytometry was used to detect the apoptosis induced by hesperidin on AGS cells, the level of reactive oxygen species, and the addition of NAC Post-apoptosis changes; Western blotting was used to detect the expression of apoptosis-related proteins and signaling pathway-related proteins. Results: MTT assay showed that hesperidin had a good inhibiting effect on AGS cells. After treated with hesperidin, AGS cells showed apoptosis such as nuclear condensation and cell shrinkage. Annexin V-FITC/PI double staining and flow cytometry showed that hesperidin can induce mitochondrial dependent apoptosis of AGS cells and increase the level of intracellular reactive oxygen species. After pretreatment of NAC, hesperidin induced apoptosis inhibition. The results of Western blotting showed that the expression of p-JNK, p-p38, Bad, cleaved Caspase-3, and cleaved PARP increased, and the expression of anti-apoptotic proteins p-ERK and Bcl-2 decreased, which indicated that hesperidin activated the MAPK signaling pathway and mitochondria-dependent apoptosis in AGS cells. Conclusion: Hesperidin has a good killing effect on human gastric cancer AGS cells, and induces mitochondria-dependent apoptosis in AGS cells by increasing the level of reactive oxygen species in AGS cells and regulating MAPK signaling pathway.
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Objective: To identify the potential target and mechanisms of hesperidin in MCF-7 estrogen receptor-positive breast cancer cells using bioinformatics approaches. Methods: Gene expression profiles were accessed from public database GSE85871. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was carried out with Database for Annotation, Visualization and Integrated Discovery. The protein-protein interaction network was analyzed by STRING-DB and visualized by Cytoscape. Transcription factor regulatory networks were constructed from TRED, TRRUST, RegNetwork and visualized by Cytoscape. Drug association analysis was conducted by WebGestalt. Results: GO and KEGG pathway enrichment analysis revealed biological processes, cellular components and molecular functions that were related to cancer and estrogen signaling pathways. KEGG pathway enrichment analysis of the genes in transcription factor-differential expression genes regulatory network showed regulation of cancer, estrogen signaling pathways, epidermal growth factor receptor tyrosine kinase inhibitor resistance, and endocrine resistance. Moreover, drug association analysis revealed that hesperidin affected the expression of the same gene as raloxifene. Conclusions: Hesperidin targets estrogen receptor signaling in estrogen receptor-positive breast cancer cells. Results of this study could trace the molecular mechanism of hesperidin in estrogen receptor-positive breast cancer cells and integrative bioinformatics analysis could accelerate drug discovery and development.
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OBJECTIVE： To establish the fingerprint of Maizao yishen granules， and to provide scientific basis for its further development. METHODS： HPLC method was adopted to establish the fingerprint by using 10 batches of Maizao yishen granules sa samples. The determination was performed on Venusil XBP C18（L） column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid （gradient elution） at the flow rate of 1→0.7 mL/min at 7-10 min， 0.7→1 mL/min at 10-15 min and 1 mL/min at the rest of time. The detection wavelengths were set at 284 nm （0-7 min）， 330 nm （7-32 min） and 360 nm （32-45 min）. The column temperature was 25 ℃， and sample size was 10 μL. The fingerprint of Maizao yishen granules was established， and the similarity evaluation was performed by using “Similarity Evaluation System of TCM Chromatographic Fingerprints” （2004 A edition） software. Then， the common peaks were assigned and identified by comparing reference substance and control medicinal materials. RESULTS： The precision， stability （24 h） and repeatability of the methodological investigation were all good [RSD values of relative retention time and relative peak area of each chromatographic peak were less than 3％ （n＝6）]. The similarity of 10 batches of samples were all above 0.900. Seventeen common peaks were identified， of which common peak 1 and 6 came from Semen Raphani； common peak 7， 9， 14， 15 and 16 from Citrus reticulata； common peak 5， 10， 11， 12 and 13 came from Glycyrrhiza uralensis； common peak 2 came from C. reticulata， G. uralensis and Ziziphus jujuba； peak 3 came from G. uralensis and Semen Raphani； peak 8 came from Hordeum vulgare and Semen Raphani； peak 4 and 17 came from C. reticulata and G. uralensis. Peak 1 was identified as hesperidin and the peak 9 was identified as sinapine. CONCLUSIONS： Established fingerprint of Maizao yishen granules is accurate and reliable， and can be used for quality control of Maizao yishen granules.
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@#AIM: To evaluate hesperidin's inhibitory effect on the proliferation of human pterygium fibroblasts(HPF)cultured <i>in vitro</i> and its influence on the expression of cyclin D.<p>METHODS: The fresh tissue of human pterygium was cultivated by adherent cell culture <i>in vitro </i>and adherent cells were appraised by immune fluorescence staining. HPF cells were treated with hesperidin(24μmol/L, 48μmol/L, 64μmol/L, 72μmol/L, 96μmol/L, 120μmol/L)and MMC(1.5μmol/L, 7.5μmol/L and 30.0μmol/L). The inhibition rate of cell proliferation was detected by MTT assay 24h, 48h and 72h after treatment, and appropriate concentration and time were selected. The relative expression of cyclin D in HPF was detected by Western blot.<p>RESULTS: When HPF were treated respectively with hesperidin(48μmol/L, 72μmol/L)and MMC(7.5μmol/L)for 48h, Western blot results showed the relative expressions of cyclin D in blank control group(normal culture), MMC group, hesperidin(48μmol/L)group and hesperidin(72μmol/L)group to be 1.20±0.02, 0.60±0.03, 0.54±0.02, 0.45±0.07(<i>F</i>=73.025, <i>P</i>=0.001)respectively. The relative expressions of cyclin D in MMC group and hesperidin group were lower than that of blank control group(<i>P</i><0.05); while the relative expressions of cyclin D in MMC group and hesperidin(48μmol/L, 72μmol/L)group showed no significant difference(<i>P</i>>0.05).<p>CONCLUSION:Hesperidin can inhibit the proliferation of HPF by reducing the relative expression of cyclin D.
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Objective: To identify the potential target and mechanisms of hesperidin in MCF-7 estrogen receptor-positive breast cancer cells using bioinformatics approaches. Methods: Gene expression profiles were accessed from public database GSE85871. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was carried out with Database for Annotation, Visualization and Integrated Discovery. The protein-protein interaction network was analyzed by STRING-DB and visualized by Cytoscape. Transcription factor regulatory networks were constructed from TRED, TRRUST, RegNetwork and visualized by Cytoscape. Drug association analysis was conducted by WebGestalt. Results: GO and KEGG pathway enrichment analysis revealed biological processes, cellular components and molecular functions that were related to cancer and estrogen signaling pathways. KEGG pathway enrichment analysis of the genes in transcription factor-differential expression genes regulatory network showed regulation of cancer, estrogen signaling pathways, epidermal growth factor receptor tyrosine kinase inhibitor resistance, and endocrine resistance. Moreover, drug association analysis revealed that hesperidin affected the expression of the same gene as raloxifene. Conclusions: Hesperidin targets estrogen receptor signaling in estrogen receptor-positive breast cancer cells. Results of this study could trace the molecular mechanism of hesperidin in estrogen receptor-positive breast cancer cells and integrative bioinformatics analysis could accelerate drug discovery and development.
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Hesperidin, a natural compound, suppresses the epithelial-to-mesenchymal transition through the TGF-ß1/Smad signaling pathway. However, studies on the detailed effects and mechanisms of hesperidin are rare. The present study showed that, for A549 alveolar epithelial cells, the anti-proliferative effects of hesperidin occurred in a dose-dependent manner, with an IC50= 216.8 µM at 48 h. TGF-ß1 was used to activate the Smad signaling pathway and induce the epithelial to mesenchymal transition in cells. Treatment with hesperidin or SB431542 was used for antagonism of Smad pathway activation. Hesperidin inhibited the increase in É-SMA and Col1É-1 and the decrease in E-cadherin in a dose-dependent manner from concentration of 20 µM to 60 µM, as assessed by both ELISA and Western blotting assays; however, there was no significant effect on cellular morphological alterations. Moreover, the Western blotting assay showed that, in the cytoplasm, hesperidin and SB431542 had no significant effect on the protein expression of Smad 2, 3, 4, or 7 as well as 2/3. However, 60 µM hesperidin and SB431542 significantly decreased p-Smad2/3 protein expression. From the above results, it is concluded that hesperidin can partly inhibit the epithelial to mesenchymal transition in human alveolar epithelial cells; the effect accounts for the blockage of the phosphorylation of Smad2/3 in the cytoplasm rather than a change in Smad protein production in the cytoplasm