Crc1,6RhaT is involved in the synthesis of hesperidin of the main bioactive substance in the Citrus reticulata ‘Chachi' fruit

Hesperidin is a dihydroflavonoids, accounting for more than 50% of the total flavonoids in Citri Reticulatae Pericarpium(CRP) of traditional Chinese medicine. It is an effective antioxidant and free radical scavenger that has anti-inflammatory, antiviral and hypoglycemic properties.The latest studies reported that hesperidin has a potential for novel coronavirus resistance. However, little is known about the synthesis regulation and accumulation site of hesperidin in plants. In this study, hesperidin synthase gene Crc1,6RhaT was cloned, and the protein can be completely transformed flavanone-7-O-glucoside into hesperidin in vitro and in vivo. Studies on biological characteristics of ovary walls and exocarps showed that the relative expression levels of the Crc1,6RhaT gene and protein decreased gradually with the development of citrus fruits, and the relative content of hesperidin firstly increased, then sequentially decreased. In situ hybridization results further revealed that Crc1,6RhaT transcription was mainly concentrated in the secretory cavity cells, which are revealed to be the site of flavonoid synthesis.Immunocytochemistry localization results showed that the Crc1,6RhaT was mainly located in the endoplasmic reticulum, nucleus and vacuole of secretory cells. We inferred that the Crc1,6RhaT was synthesized in the endoplasmic reticulum, then was transported into the vacuoles through enlarged vesicles at the end of the endoplasmic reticulum. Our results not only revealed that Crc1,6RhaT may be involved in the synthesis of hesperidin of the main bioactive substance in the medicinal plant Citrus reticulata ‘Chachi' fruit, but also provided novel insights into the main subcellular sites of hesperidin biosynthesis in vacuoles.

Hesperidin ameliorates H_(2)O_(2)-induced bovine mammary epithelial cell oxidative stress via the Nrf2 signaling pathway

Background Hesperidin is a citrus flavonoid with anti-inflammatory and antioxidant potential. However, its protective effects on bovine mammary epithelial cells(b MECs) exposed to oxidative stress have not been elucidated.Results In this study, we investigated the effects of hesperidin on H_(2)O_(2)-induced oxidative stress in b MECs and the underlying molecular mechanism. We found that hesperidin attenuated H_(2)O_(2)-induced cell damage by reducing reactive oxygen species(ROS) and malondialdehyde(MDA) levels, increasing catalase(CAT) activity, and improving cell proliferation and mitochondrial membrane potential. Moreover, hesperidin activated the Keap1/Nrf2/ARE signaling pathway by inducing the nuclear translocation of Nrf2 and the expression of its downstream genes NQO1 and HO-1, which are antioxidant enzymes involved in ROS scavenging and cellular redox balance. The protective effects of hesperidin were blocked by the Nrf2 inhibitor ML385, indicating that they were Nrf2 dependent.Conclusions Our results suggest that hesperidin could protect b MECs from oxidative stress injury by activating the Nrf2 signaling pathway, suggesting that hesperidin as a natural antioxidant has positive potential as a feed additive or plant drug to promote the health benefits of bovine mammary.

Hesperidin attenuates arsenic trioxide-induced cardiac toxicity in rats

Objective:To explore the cardioprotective effect of hesperidin against arsenic trioxide-induced cardiac toxicity in rats.Methods:Cardiac toxicity was induced by oral administration of 4 mg/kg arsenic trioxide for 30 days.Hematological,biochemical,electrocardiography,echocardiography,and histopathological examinations were performed.Results:Hesperidin decreased the neutrophil-to-lymphocyte ratio,calcium,creatine kinase-myoglobin binding,lactate dehydrogenase,IL-6,and lipid peroxidation,as well as increased sodium and potassium concentration and superoxide dismutase and catalase activity in arsenic trioxide-intoxicated rats.Moreover,it reduced peak systolic velocity and end-diastolic velocity while increasing heart rate.Arsenic trioxide-induced histopathological damage to cardiac tissue was prominently alleviated by hesperidin treatment.Conclusions:Hesperidin attenuates arsenic trioxide-induced cardiac toxicity in rats.Therefore,it can be further explored as a cardioprotective agent.

Impact of citrus fruit and hesperidin intake on multiple health outcomes:An umbrella review

Citrus fruits are rich sources of several biologically active flavonoids such as hesperidin,naringin,and polymethoxylated flavones.We evaluated the evidence of associations between citrus fruit or hesperidin intake and multiple health outcomes.An umbrella review was conducted for studies performed in humans.Overall,246 articles were initially identified by searching in 4 databases.Twenty-two meta-analyses and systematic reviews with 28 health outcomes met the inclusion criteria.Citrus fruit intake had beneficial effects on all-cause mortality(relative risk[RR].0.90;95%confidence interval[95%CI],0.86 to 0.94),cardiovascular diseases(RR,0.78;95%CI,0.66 to 0.92),coronary heart disease(RR,0.91;95%CI,0.86 to 0.96),stroke(RR,0.74;95%CI,0.65 to 0.84),type 2 diabetes mellitus(RR,0.85;95%CI,0.78 to 0.92),and several cancers.Dose-response analyses indicated that each 100-g/d increase in citrus fruit intake could reduce the risks of all-cause mortality by 6%(RR,0.94;95%CI,0.88 to 1.00),stroke by 22%(RR,0.78;95%CI,0.69 to 0.90),and cardia gastric cancer by 40%(RR,0.60;95%CI,0.44 to 0.83).Citrus fruit intake also had beneficial effects on the lipid profile and body weight control(weighted mean difference,−1.28;95%CI,−1.82 to−0.74).Grapefruits could reduce the systolic blood pressure(weighted mean difference,−2.43,95%CI,−4.77 to−0.09).Hesperidin supplementation significantly improved inflammation.Citrus fruit intake was generally safe and beneficial for multiple health outcomes in humans.However,grapefruit and pomelo juice may affect the bioavailability of various medications,so care should be exercised before increasing the intake of these fruits or their juices.

Determination of Hesperidin in Citrus Peel by HPLC

[Objective] This study aimed to measure the hesperidin content in citrus peel by high performance liquid chromatography, to provide a scientific basis for quality control and identification. [Method] The hesperidin was extracted with alkaline solution at 70 ~C and pH 6-7, and the purity of hesperidin was determined by HPLC. [Result] The formula for the regression line was Y=466,097Xq3.415 0 （r=0.999 6）, identify- ing the relationship between hesperidin concentration and peak area, and the linear range was 0.2-1.4μg. The hesperidin solution was stable within 24 h at room temperature. The average recovery rate of hesperidin was 98.41%. [Conclusion] The HPLC method is rapid, simple, and with good linear relationship, can be used for routine analysis of hesperidin.

Hesperidin as a preventive resistance agent in MCF-7 breast cancer cells line resistance to doxorubicin

Objective:To evaluate of hesperidin to overcome resistance of doxorubicin in MCF-7 resistant doxorubicin cells(MCF-7/Dox)in cytotoxicity apoptosis and P-glycoprotein(Pgp)expression in combination with doxorubicin.Methods:The cytotoxic properties.50%inhibition concentration(IC_(50))and its combination with doxorubicin in MCF-7 cell lines resistant to doxorubicin(MCF-7/Dox)cells were determined using MTT assay.Apoptosis induction was examined by double staining assay using ethidium bromide-acridine orange.Immunocytochemistry assay was performed to determine the level and localization of Pgp.Results:Single treatment of hesperidin showed cytotoxic activity on MCF-7/Dox cells with IC_(50)value of 11μmol/L.Thus,combination treatment from hesperidin and doxorubicin showed addictive and antagonist effect(CI>1.0).Hesperidin did not increase the apoptotic induction,but decreased the Pgp expressions level when combined with doxorubicin in low concentration.Conclusions:Hesperidin has cytotoxic effect on MCF-7/Dox cells with IC_(50)of 11μmol/L.Hesperidin did not increased the apoptotic induction combined with doxorubicin.Cochemotherapy application of doxorubicin and hesperidin on MCF-7/Dox cells showed synergism effect through inhibition of Pgp expression.

Chromatographic fingerprint analysis of Fructus Aurantii Immaturus by HPLC-DAD and chemometric methods

An efficient method for quality control of Fructus Aurantii Immaturus (FAI),a famous traditional Chinese medicine (TCM) was established. A simple and reliable high-performance liquid chromatography-photodiode array detector (HPLC-DAD) procedure coupled with chemometric methods was developed for fingerprint analysis,qualitative analysis and quantitative determination of this herb. In qualitative and quantitative analyses,heuristic evolving latent projection (HELP) method was employed to resolve the overlapping peaks of the tested samples. Two bioactive components,namely hesperidin and naringin,are confirmed and determined,together with four flavonoids compounds tentatively identified including two new ones. From fingerprint analysis,the fingerprint data were processed with correlation coefficients for quantitative expression of their similarity and dissimilarity. The developed method based on an integration of chromatographic fingerprint and quantitative analysis is scientific,and the obtained results can be applied to the quality control of herb medicine.

Quantitative estimation of hesperidin by HPTLC in different varieties of citrus peels

Objective:To develop a simple,selective,sensitive and accurate high-performance thin layer chromatography(HPTLC)method to determine the quantity of hesperidin in different varieties of citrus fruits.Methods:The method was carried out in aluminum-backed silica gel 60 F_(254)plates with ethyl acetate-methanol-water 15:3:2(%,v/v)as mobile phase.Results:A compact band was obtained for hesperidin at R_f value of(0.40±0.04).The calibration plot was linear in the range of 100-800 ng/spot of hesperidin and the correlation coefficient of 0.9986was indicative of good linear dependence of peak area on concentration.Limit of detection(8.87ng/spot),limit of quantification(23.21 ng/spot),accuracy(less than 2%)and recovery(ranging from98.55-99.38)were found satisfactory.Conclusions:The method developed can be used for routine analysis of hesperidin in crude drug as well as in herbal and pharmaceutical dosage form containing citrus fruits as an ingredient.

Determination of Hesperidin and Limonin Levels as Part of the Defense Mechanism of Some Lemon Varieties [Citrus limon （L,） Burm f.] against Mal Secco Disease [Phoma tracheiphila （Petri） Kanc,et Ghik,]

In this paper, it was aimed to identified and quantified hesperidin and limonin compounds using HPLC （High Performance Liquid Chromatography） techniques against to developing of mal secco disease caused by Phoma tracheiphila. Six citrus lemon varieties （Meyer, Kiitdiken, Enterdonato, Yediveren, Sweet lemon and Euroka） were infected by P. tracheiphila and artificial inoculation were applied in vivo conditions. Before and after inoculation, leaf, branch and stem samples were taken from each lemon varieties. The results show that the amount of hesperidin and limonin concentration was increased after the inoculations at various levels based on the lemon cultivars. Various concentrations （1, 5, 10, 25, 50, 75, 100 ppm） of hesperidin and limonin compounds were also tested under in vitro conditions to compare response of P. tracheiphila development. According to the results, hesperidin and limonin compounds play an important role against to P. tracheiphila development and Sweet Lemon variety was found to be the most resistance both observation and HPLC results.

Screening and identification of bioactive compounds from citrus against non-structural protein 3 protease of hepatitis C virus genotype 3a by fluorescence resonance energy transfer assay and mass spectrometry

BACKGROUND Hepatitis C virus genotype 3a(HCV G3a)is highly prevalent in Pakistan.Due to the elevated cost of available Food and Drug Administration-approved drugs against HCV,medicinal natural products of potent antiviral activity should be screened for the cost-effective treatment of the disease.Furthermore,from natural products,active compounds against vital HCV proteins like non-structural protein 3(NS3)protease could be identified to prevent viral proliferation in the host.AIM To develop cost-effective HCV genotype 3a NS3 protease inhibitors from citrus fruit extracts.METHODS Full-length NS3 without co-factor non-structural protein 4A(NS4A)and codon optimized NS3 protease in fusion with NS4A were expressed in Escherichia coli.The expressed protein was purified by metal ion affinity chromatography and gel filtration.Citrus fruit extracts were screened using fluorescence resonance energy transfer(FRET)assay against the protease and polyphenols were identified as potential inhibitors using electrospray ionization-mass spectrometry(MS)/MS technique.Among different polyphenols,highly potent compounds were screened using molecular modeling approaches and consequently the most active compound was further evaluated against HCV NS4A-NS3 protease domain using FRET assay.RESULTS NS4A fused with NS3 protease domain gene was overexpressed and the purified protein yield was high in comparison to the lower yield of the full-length NS3 protein.Furthermore,in enzyme kinetic studies,NS4A fused with NS3 protease proved to be functionally active compared to full-length NS3.So it was concluded that co-factor NS4A fusion is essential for the purification of functionally active protease.FRET assay was developed and validated by the half maximal inhibitory concentration(IC50)values of commercially available inhibitors.Screening of citrus fruit extracts against the native purified fused NS4A-NS3 protease domain showed that the grapefruit mesocarp extract exhibits the highest percentage inhibition 91%of protease activity.Among the compounds identified by LCMS analysis,hesperidin showed strong binding affinity with the protease catalytic triad having S-score value of-10.98.CONCLUSION Fused NS4A-NS3 protease is functionally more active,which is effectively inhibited by hesperidin from the grapefruit mesocarp extract with an IC50 value of 23.32μmol/L.

Anticataractogenic effect of hesperidin in galactose-induced cataractogenesis in Wistar rats

AIM: To explore the anticataractogenic potential of hesperidin, a flavanone, in galactose-induced cataractogenesis.METHODS: In this study, cataract was induced by administering galactose enriched food in a set of rats. Effect of different dosages of hesperidin(25, 50 and 75 mg/kg body weight) were administered simultaneously with galactose in prevention of cataract was determined in another set. In both sets of animals, the levels of peroxidation, oxidants(NO and OH), antioxidants(enzymatic: Superoxide dismutase, catalase, glutathione S-transferase, GPx and non-enzymatic: Reduced glutathione, vitamin E), aldose reductase and sorbitol were determined in the eye lens. In addition, glucose and lipid peroxidation levels were also tested in serum. The quantitative changes in lens inducible nitric oxide synthase(iN OS) and its expression were also determined using Western blot and real-time polymerase chain reaction analyses.RESULTS: Galactose enriched food produced cataract in both the eye lens as a sequel to elevated serum glucose. Simultaneous administration of hesperidin not only reduced serum glucose but also prevented cataract development, through reduced levels of reactive oxygen species(NO and OH) and i NOS expression as well as elevated enzymic and non-enzymic antioxidants were observed in the eye lens.CONCLUSION: These results indicate the preventive effect of hesperidin against cataract in hyperglycemic rats.

Targets and molecular mechanisms of a citrus flavonoid, hesperidin, against luminal breast cancer cells: an integrative bioinformatics analysis

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,Reg Network and visualized by Cytoscape.Drug association analysis was conducted by Web Gestalt.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.

Myricetin and Hesperidin Inhibit Cerebral Thrombogenesis and Atherogenesis in <i>Apoe^-/-</i>and <i>Ldlr^-/-</i>Mice

Flavonoids have been reported to possess strong antioxidant activities that moderate endothelial dysfunction and demonstrate protective effects on cardiovascular disease. Our previous studies confirmed that flavonoids, including hesperidin, naringin and nobiletin, inhibited thrombogenesis and hypertension in stroke prone spontaneously hypertensive rats (SHRSP) by protecting the endothelium from the adverse effects of free radical formation. We have now further investigated the protective effects of myricetin and hesperidin on cerebral thrombosis and atherogenesis in apolipoprotein E (apoE) and lowdensity lipoprotein receptor (LDLR) deficient (Apoe-/- and Ldlr-/- double knockout) mice. Three groups of mice were fed high fat diet alone and high fat diet mixed with myricetin (100 mg/kg/day and 200 mg/kg/day) or glucosyl hesperidin (G-hesperidin;250 mg/kg/day and 500 mg/kg/day) for 8 weeks. There were no differences in body weight related to administration of the flavonoids. Thrombotic tendency was assessed using a He-Ne laser technique in the murine cerebral pial vessels. In addition, atherogenesis was quantified histologically after dissection of the aorta from each mouse and staining with Oil Red O solution. The percentages of stained area to whole area of dissected aorta were calculated as indices of anti-atherogenic activity. Both myricetin and G-hesperidin significantly inhibited thrombogenesis in vivo and significantly inhibited atherogenesis compared to control mice (p < 0.001). These findings demonstrated that daily intake of myricetin and hesperidin suppressed the development of atherogenesis and thrombogenesis, possibly associated with the potent antioxidant effects of the flavonoids.

Citrus Peel Ethanol Extract Inhibits the Adipogenesis Caused from High Fat-Induced DIO Model

Background: Flavonoids are multi-functional bioactive compounds that have been used as natural compounds against various diseases. Citrus fruit is an important source for bioactive flavonoids with potential anti-obesity benefits. Methods: To determine the anti-obese effects of citrus peel, a 45% high fat diet-induced obesity (DIO) model using C57BL/6 mice was prepared for 10 weeks and then treated orally for 12 weeks with ethanol extracts of citrus peel (300 mg/kg, CP). CP was compared with normal chow diet (C), high fat diet (HF), and the anti-obesity drug orlistat (30 mg/kg, O) as a positive control. HF caused increases in lipid accumulation, body weight gain, and hepatic toxicity compared with the C group. Results: CP treatment reduced body weight gain and decreased epididymal fat, mesenteric fat, and plasma and hepatic TG levels in a similar manner as O treatment. Besides, CP was comparatively more effective than O at increasing high density lipoprotein cholesterol (HDL-c) while reducing hepatic toxicity, which is caused by HF. Fat accumulation in adipose tissue was decreased by CP treatment because of up-regulation of specific lipolysis enzymes such as HSL and AMPK and down-regulation of adipogenesis related genes such as C/EBPα and ACC. The proinflammatory cytokines, TNF-α and IL-6, which are the key factors for regulation of inflammation, were significantly decreased by CP. Conclusion: CP may be a potential natural source for new anti-obesity candidate because of its inhibitory effect on fat synthesis-related inflammation and its positive effect on lipolysis activation.

Mechanism of hesperidin improving myocardial ischemia/reperfusion injury in type 2 diabetic rats through SIRT1/Nrf2/HO-1 signaling pathway

Objective:To observe the protective effect of hesperidin on myocardial ischemia/reperfusion injury in type 2 diabetes mellitus and its effect on SIRT1/Nrf2/HO-1 signaling pathway.Methods:50 Sprague-Dawley(SD)rats were randomly assigned to the normal control group(NC),model group,ischemia-reperfusion group(IR),hesperidin group,SIRT1 inhibitor group and hesperidin plus SIRT1 inhibitor group.In addition to NC,the rats in the remaining groups were replicated by intraperitoneal of high-fat diet combined with injection of streptozotocin for type 2 diabetic rats.After then,the myocardial ischemia/reperfusion injury(MIRI)rat model was established by LAd for 30 minutes with 2 hours reperfusion.He staining was used to observe the pathological changes of myocardial tissue,and the levels of serum LDH,CK-MB and SOD,GSH and MDA in myocardial tissue were detected by kit methods,and the expression abundance of related proteins in 4-HNE and SIRT1/Nrf2/HO-1 signal pathway were detected by immunohistochemistry and Western blot;Results:Hesperidin could significantly inhibit cardiomyocyte necrosis and inflammatory cell infiltration,reduce LDH activity,CK-MB and MDA level,and increase SOD activity,GSH and 4-HNE level,the differences were statistically significant when compared with IR group(P<0.01).In addition,compared with the ischemia-reperfusion group,the expressions of SIRT1,Nrf2 and HO-1 proteins in hesperidin group were significantly up-regulated,the differences were statistically significant(P<0.01);Conclusion:Hesperidin inhibits oxidative stress by activating SIRT1/Nrf2/HO-1 signaling pathway,and play a protective effect of myocardial ischemia reperfusion injury in diabetic rats.

Neuroprotective Effects of Hesperidin and Benfotiamine against Paraquat Induced Spinal Cord Neurotoxicity in Adult Male Albino Rats

Introduction: There are extensive people exposures to paraquat (PQ) herbicide resulting in human health hazards. Aim of the Work: To compare the beneficial neuroprotective effects of hesperidin and benfotiamine on paraquat (PQ)-induced spinal cord neurotoxic effects in rats. Materials and Methods: Rats were divided into 4 groups as following: control, paraquat (PQ 20.8 mg/kg, oral gavage (e.g.)), paraquat + benfotiamine (50 mg/kg, oral gavage (e.g.)) and paraquat + hesperidin (40 mg/kg, oral gavage (e.g.)). PQ is given as the previous dose. Rats are treated 6 days per week. Results: There was a significant increased mean value of malondialdehyde associated with a significant reduction in the content of reduced glutathione and antioxidant enzymes activities associated with a significant increase in Serum phosphorylated neurofilament-H, neurospecific enolase and s100 levels were recorded and significant spinal cord histopathological changes in paraquat treated group as compared to their corresponding values in the control group and immunohistochemical examination confirmed these results. Upon supplementation with benfotiamine and hesperidin to paraquat treated rats, there was a significant decrease in the mean values of malondialdehyde associated with a marked increase in the content of reduced glutathione and antioxidant enzymes activities associated with a significant decrease in Serum phosphorylated neurofilament-H, neurospecific enolase and s100 levels were also recorded with significant improvement of spinal cord architecture when compared with the paraquat treated group. Conclusion: The use of benfotiamine and hesperidin produced a significant protection against all of the above-mentioned changes.

Hesperidin Reduces Cisplatin-lnduced DNA Damage in Bone Marrow Cells of Mice

Hesperidin is a bioflavonoid abundantly found in citrus fruits and displays chemoprotective effects against DNA （deoxyribonucleic acid） damage, however there are few reports about hesperidin effects against cisplatin-DNA damage induction. The aim of this work was to evaluate hesperidin antimutagenicity against cisptatin-DNA damage. （1） The antimutagenicity of hesperidin was assayed by bone marrow of mice in vivo using the micronucleus test. Hesperidin pre-treatment protocol reduced the frequency of MNPCE （micronucleated polychromatic erythrocytes） and the dose of 100 mg·kg-1 was highest efficiency, with 65.24% of damage reduction. In the simultaneous treatment protocol, the dose of 200 mg·kg-1 exhibited a more effective reduction of MNPCE, with 94.01% of damage reduction. （2） Hesperidin was also effective in reducing the MNPCE frequency in the post-treatment protocol for all doses, with 77.48%, 82.13% and 90.08% of damage reduction at the doses of 100, 200 and 400 mg·kg-1, respectively. From the study, it can be concluded that hesperidin was able to promote the reduction of micronuclei frequency and DNA damage induced by cisplatin. Hesperidin is a powerful antioxidant compound and its chemoprotective effects on DNA may occur due to its association with the antioxidant cell system which is responsible for eliminate free radicals generated by chemical harmful to DNA.

The Protective Role of β-Carotene and Hesperidin on Some Hematological and Myocardial Measurements against Imidacloprid Toxicity in Albino Rats

Neonicotinoids including IM （Imidacloprid） are widely used as plant systemic insecticides. Several studies have indicated that pesticide toxicity may be associated with the enhanced production of ROS （reactive oxygen species）. Both β-carotene （I3C） and hesperidin （H） have an antioxidant property and quench free radicals. This study aimed to clarify the protective role of β-carotene and hesperidin as natural antioxidants on IM induced toxicity in hematological parameters and markers of cardiac muscle activity in male albino rats. The treatment of rats with IM showed a significant decrease in hemoglobin （Hb %）, MCH （mean corpuscular hemoglobin）, MCHC （mean corpuscular hemoglobin concentration）, HCT （hematocrit） values and RBCs count comparing with control group. On the other hand, MCV （mean corpuscular volume）, WBCs （white blood cells） and Pits （platelets） count pronounced a significant increase in IM group comparing to control. Also, αFP （plasma alpha fetoprotein）, CEA （carcinoembryonic antigen）, CK （creatine kinase）, CK-MB （creatine phosphokinase myocardial band） and LDH （lactate dehydrogenase） clarify a significant increase in IM group comparing to control. Both β-carotene and hesperidin mitigate the deleterious effects of IM on previous parameters. β-Carotene and hesperidin may protect hematopoietic system and heart muscle against toxicity of IM. These improvements of the results clarify the protective effect of the used antioxidants. Conclusion： β-carotene and hesperidin, natural antioxidants, have a protective effect against IM evoked hematological and biochemical changes.

Antidepressant-Like Effects of Hesperidin in Animal Model of Post-Traumatic Stress Disorder

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 significantly improved depression-like behaviors in the FST(P<0.05),increased the number of lines crossed in the central zone of the OFT(P<0.01),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 flavonoid may be an effective medicine for PTSD.

Reliability of online near-infrared spectroscopy analysis of the ex-traction process of Huatan Qushi decoction for phlegm-dampness constitution

OBJECTIVE: To establish a quantitative model for the online analysis of hesperidin content in the extraction process of Huatan Qushi decoction for individuals with phlegm-dampness constitution by using near-infrared(NIR) spectroscopy and partial least squares(PLS) methods.METHODS: The reference content of hesperidin, an effective ingredient of Huatan Qushi decoction,was determined using high-performance liquid chromatography(HPLC). The modeling band was selected using synergy interval PLS(Si PLS) algorithms, and a PLS model demonstrating the relationship between NIR predictive values and HPLC reference values was established. The optimal modeling route was selected through global optimization of process trace parameters.RESULTS: The root mean square error of cross-validation was 9.8410. The coefficients of determination of the calibration set and prediction set were0.9860(R2 cal) and 0.9458(R_(cal)~2), respectively. The residual predictive deviation was 3.0115. The parameters demonstrated high predictive capability and reliability. The band was selected via the Si PLS method. There were seven latent variables.CONCLUSION: This study provides a favorable comprehensive evaluation method for variable selection and quality control of the Traditional Chinese Medicine extraction process.