Protective Effect and Autophagy Mechanism of Lycium barbarum Polysaccharides on Retinal Pigment Epithelial Cells Under High-Glucose Conditions

Objective:To study the effects of Lycium barbarum polysaccharide(LBP)on the proliferation,apoptosis,and autophagy of retinal pigment epithelial(RPE)cells cultured under high-glucose conditions.Methods:The ARPE-19 cell line was randomly divided into a control group(normally cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12[DMEM/F-12]medium),a high-glucose group(HG;50 mmol/L glucose added to DMEM/F-12 medium),and a HG+LBP group(incubated in DMEM/F-12 medium containing 1 mg/mL LBP for 24 h,and then treated with 50 mmol/L glucose for 24 h).Following Ad-mCherry-GFP-LC3B infection,cell proliferation,apoptosis,mammalian target of rapamy-cin(mTOR)expression,and autophagic flux were determined by Cell Counting Kit-8(CCK-8),AnnexinV-APC/7-AAD Apoptosis Detection Kit,Western blot,and laser confocal microscopy,respectively.Results:The proliferation rate of ARPE-19 cells in the HG group was significantly lower than that in the control group(P<0.05),while the proliferation rate of ARPE-19 cells in the HG+LBP group was significantly higher than that in the HG group(P<0.05).The apoptosis rate of ARPE-19 cells in the HG group was significantly higher than that in the control group(P<0.05),while the apoptosis rate of ARPE-19 cells in the HG+LBP group was significantly lower than that in the HG group(P<0.05).The relative expression of phosphorylated mTOR(p-mTOR)of ARPE-19 cells in the HG group was significantly lower than that in the control group(P<0.05),with enhanced autophagic flux;when compared with the HG group,the HG+LBP group had significantly higher expression of p-mTOR(P<0.05),with diminished autophagic flux.Conclusion:LBP has a protective effect on RPE cells with high glucose-induced injury,and its mechanism may be related to LBP inhibition of high glucose-induced abnormal autophagy.

Potential role of Lycium barbarum polysaccharides in glaucoma management:evidence from preclinical in vivo studies

In recent years,the pharmacological benefits of herbal extracts have been revisited for their potential neuroprotective effects in glaucoma.The polysaccharides extracted from the fruits of Lycium barbarum L.,or Lycium barbarum polysaccharides,exert their anti-aging effect through reducing oxidative stress,modulating the immune response,enhancing neuronal responses,and promoting cytoprotection.The therapeutic efficacy of Lycium barbarum polysaccharides in preserving retinal ganglion cells and their functions was demonstrated in a range of experimental models of optic neuropathies.These include the acute and chronic ocular hypertension models,the partial optic nerve transection model,and the ischemic-reperfusion injuries model.Based on these findings,Lycium barbarum polysaccharides appear to be a good candidate to be developed as a neuroprotective agent for treating multifactorial diseases.This review aims to present a comprehensive review on the latest preclinical evidence on the pre-and post-treatment benefits of Lycium barbarum polysaccharides in retinal ganglion cell neuroprotection.The possible mechanisms of Lycium barbarum polysaccharides mediating retinal ganglion cell neuroprotection will also be described.Moreover,the potential research gaps in the effective translation of Lycium barbarum polysaccharides treatment into clinical glaucoma management will be discussed.

Lycium barbarum polysaccharides protects retinal ganglion cells against oxidative stress injury

The accumulation of excessive reactive oxygen species can exacerbate any injury of retinal tissue because free radicals can trigger lipid peroxidation,protein damage and DNA fragmentation.Increased oxidative stress is associated with the common pathological process of many eye diseases,such as glaucoma,diabetic retinopathy and ischemic optic neuropathy.Many studies have demonstrated that Lycium barbarum polysaccharides(LBP)protects against oxidative injury in numerous cells and tissues.For the model of hypoxia we used cultured retinal ganglion cells and induced hypoxia by incubating with 200μM cobalt chloride(CoCl2)for 24 hours.To investigate the protective effect of LBP and its mechanism of action against oxidative stress injury,the retinal tissue was pretreated with 0.5 mg/mL LBP for 24 hours.The results of flow cytometric analysis showed LBP could effectively reduce the CoCl2-induced retinal ganglion cell apoptosis,inhibited the generation of reactive oxygen species and the reduction of mitochondrial membrane potential.These findings suggested that LBP could protect retinal ganglion cells from CoCl2-induced apoptosis by reducing mitochondrial membrane potential and reactive oxygen species.

Tissue distribution of Lycium barbarum polysaccharides in rat tissue by fluorescein isothiocyanate labeling

To date, in vivo investigations of polysaccharide’s pharmacokinetics are significantly restricted by the difficulty in their detection. This study was conducted to establish the quantitative determination of Lycium barbarum polysaccharides(LBPs) based on fluorescein isothiocyanate(FITC) pre-labeling and to investigate their tissue distribution in rat. We obtained the calibration curves linear over the range of 0.0–25 μg/m L in rat tissue samples with correlation coefficients greater than 0.99. The inter-day and intra-day precisions(RSD, %) were within 15%, and the relative recovery ranged 95.2%–102.4%, with RSD range 1.48%–9.58%, indicating that this experiment was suitable for the determination of LBPs. The fluorescence intensity was measured after 24 h storage at room temperature, 3 times of freeze-cycle and cryopreservation at –20 ℃ for 15 day, these results indicated that the stability of the samples was good. LBP-FITC was mainly absorbed by the small intestine and stomach, and mainly excreted in the urine through the kidney;this distinct difference in the tissue distribution of LBPs could be attributed to the size of these LBPs in relation to the pore sizes of the vascular beds in the kidney and liver. Results showed in this study enable us to comprehensively understand the biological effects of LBPs following its oral ingestion.

Therapeutic Uses of Lycium barbarum Polysaccharides on Idiopathic Thrombocytopenic Purpura Mice via Inhibition of Oxidative Stress

The aim of the present study was to investigate the effect of Lycium barbarum polysaccharides (LBP) on the treatment of ITP mice and to explore its mechanism. Forty idiopathic thrombocytopenic purpura (ITP) mice were divided randomly into a model control group and LBP groups I, II, III and IV. ITP mice in LBP groups I, II, III, and IV were administered LBP at four different doses (50, 100, 200 and 400 mg·kg-1·d-1, respectively) for 7 days by gavage. Blood samples were collected from the tail veins of the mice after treatment. Platelet counts were determined, and the total antioxidant status (TAS), total oxidant status (TOS) were measured with ELISA kits. The platelet count was (30.28 ± 13.42) × 109/L in the model control group, and the number of platelets in all LBP groups was higher than that in the model control group. The platelet count increased, and it reached (67.09 ± 10.81) × 109/L in LBP group I;the platelet counts in the other three groups increased significantly compared to LBP group I, and they did not differ significantly. TAS concentrations in the LBP groups were significantly increased compared to the model control group, whereas TOS concentrations were significantly decreased. Taken together, these results indicate that LBP is effective at increasing the number of platelet (PLT), and LBP may treat ITP mice via suppressing oxidative stress.

Effects of Lycium barbarum polysaccharides on the proliferation and differentiation of primary Sertoli cells in young rats

Objective:Sertoli cells(SCs)provide physical support and material supply for germ cells and participate in the formation of blood-testis barrier.The number of SCs is directly proportional to the number of germ cells.And mature SCs ensure the growth of germ cells and the production of sperm.In this study,we explored the effect and underlying mechanism of Lycium barbarum polysaccharides(LBP)on primary SCs in young rats.Methods:Primary SCs were isolated from the testis of 20-day old rats.The cells were then treated with different concentrations of LBP.Immunocytochemistry was used to detect the expression of Ki67 and the androgen receptor(AR),and western blotting was used to detect the expression of cytokeratin-18(CK-18),AR and phosphorylated Akt(Ser473)in SCs.Results:The number of SCs increased significantly after LBP treatment,and the 100 mg/mL.LBP group had 14%more cells than the control group.The expression of Ki67 in LBP treated groups also increased significantly.LBP inhibited the expression of cytokeratin 18 in SCs.Besides,LBP increased the expression of AR on SCs and promoted the activation of Akt at the ser473 phosphorylation site.Conclusion:LBP promotes the proliferation of immature SCs in young rats and also accelerates their differentiation and maturation.This seems to be associated with activation of the Akt signaling pathway via up-regulation of AR.

Lycium barbarum polysaccharides ameliorate canine acute liver injury by reducing oxidative stress,protecting mitochondrial function,and regulating metabolic pathways

The development of acute liver injury can result in liver cirrhosis,liver failure,and even liver cancer,yet there is currently no effective therapy for it.The purpose of this study was to investigate the protective effect and therapeutic mechanism of Lycium barbarum polysaccharides(LBPs)on acute liver injury induced by carbon tetrachloride(CCl_(4)).To create a model of acute liver injury,experimental canines received an intraperitoneal injection of 1 mL/kg of CCl_(4)solution.The experimental canines in the therapy group were then fed LBPs(20 mg/kg).CCl_(4)-induced liver structural damage,excessive fibrosis,and reduced mitochondrial density were all improved by LBPs,according to microstructure data.By suppressing Kelch-like epichlorohydrin(ECH)-associated protein 1(Keap1),promoting the production of sequestosome 1(SQSTM1)/p62,nuclear factor erythroid 2-related factor 2(Nrf2),and phaseⅡdetoxification genes and proteins downstream of Nrf2,and restoring the activity of anti-oxidant enzymes like catalase(CAT),LBPs can restore and increase the antioxidant capacity of liver.To lessen mitochondrial damage,LBPs can also enhance mitochondrial respiration,raise tissue adenosine triphosphate(ATP)levels,and reactivate the respiratory chain complexes I–V.According to serum metabolomics,the therapeutic impact of LBPs on acute liver damage is accomplished mostly by controlling the pathways to lipid metabolism.9-Hydroxyoctadecadienoic acid(9-HODE),lysophosphatidylcholine(Lyso PC/LPC),and phosphatidylethanolamine(PE)may be potential indicators of acute liver injury.This study confirmed that LBPs,an effective hepatoprotective drug,may cure acute liver injury by lowering oxidative stress,repairing mitochondrial damage,and regulating metabolic pathways.

Effects of Lycium barbarum polysaccharide on cytokines in adolescents with subthreshold depression:a randomized controlled study

Strong evidence has accumulated to show a correlation between depression symptoms and inflammatory responses.Moreover,anti-inflammatory treatment has shown partial effectiveness in alleviating depression symptoms.Lycium barbarum polysaccharide(LBP),derived from Goji berries,exhibits notable antioxidative and anti-inflammatory properties.In our recent double-blinded randomized placebo-controlled trial,we found that LBP significantly reduced depressive symptoms in adolescents with subthreshold depression.It is presumed that the antidepressant effect of LBP may be associated with its influence on inflammatory cytokines.In the double-blinded randomized controlled trial,we enrolled 29 adolescents with subthreshold depression and randomly divided them into an LBP group and a placebo group.In the LBP group,adolescents were given 300 mg/d LBP.A 6-week follow up was completed by 24 adolescents,comprising 14 adolescents from the LBP group(15.36±2.06 years,3 men and 11 women)and 10 adolescents from the placebo group(14.9±1.6 years,2 men and 8 women).Our results showed that after 6 weeks of treatment,the interleukin-17A level in the LBP group was lower than that in the placebo group.Network analysis showed that LBP reduced the correlations and connectivity between inflammatory factors,which were associated with the improvement in depressive symptoms.These findings suggest that 6-week administration of LBP suppresses the immune response by reducing interleukin-17A level,thereby exerting an antidepressant effect.

Lycium barbarum Polysaccharides Promotes Mitochondrial Biogenesis and Energy Balance in NAFLD Cell Model

Objective:To explore the protective effect and underlying mechanism of Lycium barbarum polysaccharides(LBP)in a non-alcoholic fatty liver disease(NAFLD)cell model.Methods:Normal human hepatocyte LO2 cells were treated with 1 mmol/L free fatty acids(FFA)mixture for 24 h to induce NAFLD cell model.Cells were divided into 5 groups,including control,model,low-,medium-and high dose LBP(30,100and 300μg/mL)groups.The monosaccharide components of LBP were analyzed with high performance liquid chromatography.Effects of LBP on cell viability and intracellular lipid accumulation were assessed by cell counting Kit-8 assay and oil red O staining,respectively.Triglyceride(TG),alanine aminotransferase(ALT),aspartate aminotransferase(AST),adenosine triphosphate(ATP)and oxidative stress indicators were evaluated.Energy balance and mitochondrial biogenesis related mRNA and proteins were determined by quantitative real-time polymerase chain reaction and Western blot,respectively.Results:Heteropolysaccharides with mannose and glucose are the main components of LBP.LBP treatment significantly decreased intracellular lipid accumulation as well as TG,ALT,AST and malondialdehyde levels(P<0.05 or P<0.01),increased the levels of superoxide dismutase,phospholipid hydroperoxide glutathione peroxidase,catalase,and ATP in NAFLD cell model(P<0.05).Meanwhile,the expression of uncoupling protein 2 was down-regulated and peroxisome proliferator-activated receptor gamma coactivator-1α/nuclear respiratory factor 1/mitochondrial transcription factor A pathway was up-regulated(P<0.05).Conclusion:LBP promotes mitochondrial biogenesis and improves energy balance in NAFLD cell model.

Lycium barbarum polysaccharide ameliorates radiation-induced brain injury by regulating gut microbiota

Objective: To determine the extent to which Lycium barbarum polysaccharide(LBP) improves60Co γ-ray radiation-induced brain injury(RIBI) by regulating the gut microbiota.Methods: The RIBI model of mice was established with the appropriate dose of60Co γ-ray to identify the changes in the body weight, behaviors, gut microbiota, and inflammatory reactions of mice. Mice were randomly divided into healthy, RIBI model, and LBP groups. The related inflammatory cytokines were determined using an enzyme linked immunosorbent assay kit. Then, 16S rRNA sequencings of feces were carried out to evaluate the differences in intestinal flora.Results: Compared with the spontaneous activity and exploratory spirit of the healthy group, those traits in the RIBI model mice in the open field significantly decreased, the freezing time in the elevated plus maze(EPM) significantly increased, and the number of times the mice discriminated the novel object was significantly lower. Hematoxylin-eosin slides showed that the main histopathological changes of RIBI occurred in the hippocampus. In addition, the diversity and relative abundances ratio of the gut bacterial phylum, order, family, and genus in the model group varied widely. Changes in Bacteroidetes,Firmicutes, and Proteobacteria were the most obvious after head radiation exposure. In comparison, LBP could accelerate the recovery of weight loss in RIBI mice. The frequency that mice entered the center of the open field, facing the open arm in the EPM, and the number of times they discriminated the novel object were significantly increased with LBP administration. LBP could also reduce the levels of inflammatory factor caused by RIBI. LBP increased the diversity and abundance of gut microbiota in RIBI model mice. In addition, LBP increased the relative abundance of Bacteroidetes but decreased the levels of Firmicutes and Proteobacteria for irradiated mice.Conclusion: LBP can improve depression and tension by regulating the composition of gut microbiota,including lowering the relative abundance of Clostridia and Burkholderiales and raising that of Lactobacillales. Thus, LBP provides a new strategy for improving the protective effects of RIBI.

Research advances on antioxidation,neuroprotection,and molecular mechanisms of Lycium barbarum polysaccharides

Lycium barbarum polysaccharides(LBPs)are the major polysaccharides extracted from L.barbarum,which is used in traditional Chinese medicine(TCM)for treating diseases.Studies have shown that LBPs have important biological activities,such as antioxidation,anti-aging,neuroprotection,immune regulation.LBPs inhibit oxidative stress,improve neurodegeneration and stroke-induced neural injury,increase proliferation and differentiation of neural stem cell,and promote neural regeneration.Here we have reviewed latest advances in the biomedical activities of LBPs and improved methods for the isolation,extraction,and purification of LBPs.Then,new discoveries to decrease oxidative stress and cellular apoptosis,inhibit aging progress,and improve neural repair in neurodegeneration and ischemic brain injury have been discussed in detail through in vitro cell culture and in vivo animal studies.Importantly,the molecular mechanisms of LBPs in playing neuroprotective roles are further explored.Lastly,we discuss the perspective of LBPs as biomedical compounds in TCM and modern medicine and provide the experimental and theoretical evidence to use LBPs for the treatment of aging-related neurological diseases and stroke-induced neural injuries.

Lycium barbarum polysaccharide-glycoprotein preventative treatment ameliorates aversive stimuli-induced depression

Previous studies have shown that Lycium barbarum polysaccharide,the main active component of Lycium barbarum,exhibits antiinflammatory and antioxidant effects in treating neurological diseases.However,the therapeutic action of Lycium barbarum polysaccharide on depression has not been studied.In this investigation,we established mouse models of depression using aversive stimuli including exposure to fox urine,air puff and foot shock and physical restraint.Concurrently,we administered 5 mg/kg per day Lycium barbarum polysaccharide-glycoprotein to each mouse intragastrically for the 28 days.Our results showed that long-term exposure to aversive stimuli significantly enhanced depressive-like behavior evaluated by the sucrose preference test and the forced swimming test and increased anxietylike behaviors evaluated using the open field test.In addition,aversive stimuli-induced depressed mice exhibited aberrant neuronal activity in the lateral habenula.Importantly,concurrent Lycium barbarum polysaccharide-glycoprotein treatment significantly reduced these changes.These findings suggest that Lycium barbarum polysaccharide-glycoprotein is a potential preventative intervention for depression and may act by preventing aberrant neuronal activity and microglial activation in the lateral habenula.The study was approved by the Jinan University Institutional Animal Care and Use Committee(approval No.20170301003)on March 1,2017.

Efficacy of Lycium barbarum polysaccharide in adolescents with subthreshold depression: interim analysis of a randomized controlled study

Subthreshold depression is a highly prevalent condition in adolescents who are at high risk for developing major depressive disorder.In preclinical models of neurological and psychiatric diseases,Lycium barbarum polysaccharide(LBP)extracted from Goji berries had antidepressant effects including but not limited to anti-oxidative and anti-inflammatory properties.However,the effect of LBP on subthreshold depression is unclear.To investigate the clinical efficacy and safety of LBP for treating subthreshold depression in adolescents,we conducted a randomized,double-blind,placebo-controlled trial(RCT)with 29 adolescents with subthreshold depression recruited at The Fifth Affiliated Hospital of Guangzhou Medical University.The participants were randomly assigned to groups where they received either 300 mg LBP(LBP group,n=15,3 boys and 12 girls aged 15.13±2.17 years)or a placebo(placebo group,n=14,2 boys and 12 girls aged 15±1.71 years)for 6 successive weeks.Interim analyses revealed that the LBP group exhibited a greater change in Hamilton Depression Scale(HAMD-24)scores relative to the baseline and a higher remission rate(HAMD-24 total score≤7)at 6 weeks compared with the placebo group.Scores on the Beck Depression Inventory-II(BDI-II),Pittsburgh Sleep Quality Index(PSQI),Kessler Psychological Distress Scale(Kessler),and Screen for Child Anxiety-Related Emotional Disorders(SCARED)were similar between the LBP and placebo groups.No side effects related to the intervention were reported in either group.These results indicate that LBP administration reduced depressive symptoms in adolescents with subthreshold depression.Furthermore,LBP was well tolerated with no treatment-limiting adverse events.Clinical trials involving a larger sample size are needed to further confirm the anti-depressive effects of LBP in adolescents with subthreshold depression.This study was approved by the Medical Ethics Committee of the Fifth Affiliated Hospital of Guangzhou Medical University(Guangzhou,China;approval No.L2019-08)on April 4,2019 and was registered on ClinicalTrials.gov(identifier:NCT04032795)on July 25,2019.

Effects of Lycium barbarum polysaccharide on the photoinduced autophagy of retinal pigment epithelium cells

AIM:To investigate the relationship between autophagy and apoptosis in photoinduced injuries in retinal pigment epithelium(RPE)cells and how Lycium barbarum polysaccharide(LBP)contributes to the increased of RPE cells to photoinduced autophagy.METHODS:In vitro cultures of human RPE strains(ARPE-19)were prepared and randomly divided into the blank control,model,low-dose LBP,middle-dose LBP,high-dose LBP,and 3-methyladenine(3MA)groups.The viability of the RPE cells and apoptosis levels in each group were tested through cell counting kit-8(CCK8)method with a flow cytometer(Annexin V/PI double staining technique).The expression levels of LC3II,LC3I,and P62 proteins were detected with the immunofluorescence method.The expression levels of beclin1,LC3,P62,PI3K,P-mTOR,mTOR,P-Akt,and Akt proteins were tested through Western blot.RESULTS:LBP considerably strengthens cell viability and inhibits the apoptosis of RPE cells after photoinduction.The PI3K/Akt/mTOR signal pathway is activated because of the upregulation of the phosphorylation levels of Akt and mTOR proteins,and thus autophagy is inhibited.CONCLUSION:LBP can inhibit the excessive autophagy in RPE cells by activating the PI3K/Akt/mTOR signaling pathways and thereby protect RPE cells from photoinduced injuries.

Effect of nano-selenium loaded with lycium barbarum polysaccharide on the proliferation of lens epithelial cells after UVB damage in vitro

AIM:To investigate the effect of nano-selenium loaded with different concentrations of lycium barbarum polysaccharide(LBP-Se NPs)on the proliferation of human lens epithelial cells(HLECs)from UV irradiation.METHODS:LBP-Se NPs were prepared and their particle size was detected.HLECs(SRA01/04)were irradiated with UVB for different time(0,10,20,30,40,50,60 min)to construct a damaged model,the survival rate of cells was determined by methylthiazol tetrazolium(MTT)assay.The 4’,6-Diamidine-2’-phenylindole dihydrochloride(DAPI)staining was used to observe the status of cell nucleus and drug entering cytoplasm through cell membrane in SRA01/04 cells after adding LBP-SENPS loaded with coumarin fluorescence agent 24 h under fluorescence microscope.SRA01/04 normal and UVB-damaged cells were treated with different amounts of LBP-Se NPs at different concentrations,cells proliferation were observed.RESULTS:The particle size of LBP-Se NPs was stable in the range of 150-200 nm.The survival rate changes with time after UVB irradiation were statistically significant.The 10 min of UVB exposure as the time was chosen to construct the cell damage model.With DAPI staining,LBPSe NPs were observed to enter the cytoplasm through the cell membrane under fluorescence inverted microscope.Cytotoxicity of SRA01/04 at different concentrations of LBPSe NPs were measured.Cell survival rate was statistically different compared with the control group.The higher the loading concentration of LBP in nano-Se drugs was,the higher the cell proliferation rate was(P<0.05).The lower the concentration of LBP-Se NPs,the higher the cell proliferation rate,showing a negative growth trend(P<0.05).The group with the highest average cell proliferation rate was 0.5μmol/L 2.0 mg/m L LBP-Se NPs(128.80%).When the 2.0 mg/m L LBP-Se NPs group was selected for cell photography,the cell density was higher at 0.5μmol/L.With the increase of concentration,SRA01/04 cells appeared more cytoplasm dehydration,cell shrinkage and apoptotic bodies,and cell density decreased.CONCLUSION:LBP-Se NPs has moderate particle size and good stability.LBP-Se NPs can protect HLECs(SRA01/04)from UVB-induced damage,and the cell proliferation rate is further increased with increasing the amount of loaded LBP and decreasing nano-selenium concentration.

Research Progress in the Application of Lycium Barbarum Polysaccharide in Ophthalmic Diseases

Lycium barbarum polysaccharides(LBP)are widely used in age-related macular degeneration,glaucoma,diabetic retinopathy,and other eye diseases.With further studies on the pharmacological action of Lycium barbarum polysaccharides in recent years,researchers have found that Lycium barbarum polysaccharides(LBP)play a protective role in the treatment of various eye diseases through anti-oxidation,scavenging oxygen free radicals,and inhibiting inflammatory reaction as well as cell senescence.This paper reviews the application of Lycium barbarum polysaccharides in ophthalmology.

A novel anticancer property of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells

Breast cancer is one of the most malignant tumors and is associated with high mortality rates among women.Lycium barbarum polysaccharide(LBP)is an extract from the fruits of the traditional Chinese herb,L.barbarum.LBP is a promising anticancer drug,due to its high activity and low toxicity.Although it has anticancer properties,its mechanisms of action have not been fully established.Ferroptosis,which is a novel anticancer strategy,is a cell death mechanism that relies on iron-dependent lipid reactive oxygen species(ROS)accumulation.In this study,human breast cancer cells(Michigan Cancer Foundation-7(MCF-7)and MD Anderson-Metastatic Breast-231(MDA-MB-231))were treated with LBP.LBP inhibited their viability and proliferation in association with high levels of ferroptosis.Therefore,we aimed to ascertain whether LBP reduced cell viability through ferroptosis.We found that the structure and function of mitochondria,lipid peroxidation,and expression of solute carrier family 7 member 11(SLC7 A11,also known as x CT,the light-chain subunit of cystine/glutamate antiporter system X_(c)^(-))and glutathione peroxidase 4(GPX4)were altered by LBP.Moreover,the ferroptosis inhibitor,Ferrostatin-1(Fer-1),rescued LBP-induced ferroptosis-associated events including reduced cell viability and glutathione(GSH)production,accumulation of intracellular free divalent iron ions and malondialdehyde(MDA),and down-regulation of the expression of x CT and GPX4.Erastin(x CT inhibitor)and RSL3(GPX4 inhibitor)inhibited the expression of x CT and GPX4,respectively,which was lower after the co-treatment of LBP with Erastin and RSL3.These results suggest that LBP effectively prevents breast cancer cell proliferation and promotes ferroptosis via the x CT/GPX4 pathway.Therefore,LBP exhibits novel anticancer properties by triggering ferroptosis,and may be a potential therapeutic option for breast cancer.

Mechanism regulating the inhibition of lung cancer A549 cell proliferation and structural analysis of the polysaccharide Lycium barbarum

The water-soluble polysaccharide LBP-1 was isolated and characterized from Lycium barbarum L.LBP-1 was mainly composed of arabinose,galactose,glucose,xylose,mannose at a molar ratio of 37.53:28.08:14.72:7.83:4.50,respectively.Based on nuclear magnetic resonance(NMR)and methylation analysis,the backbone of LBP-1 was speculated in theα-L-Ara(1→[5-α-L-Ara(1→3)-β-D-Galp-(1→]n→4)-α-D-Galp-(1[→5-α-L-Ara(1]n[→6)-β-D-Galp-(1→4)-β-D-Galp-(1→]n,and the side chains of LBP-1 were in theα-L-Ara(1→3)-β-D-Galp-(1→6 position.These results showed that LBP-1 inhibited the growth of cancer A549 cells through cell cycle arrest and apoptosis,with an IC50 value of 42.5μg/mL.In addition,LBP-1 altered the expression of Cyclin D1,Cyclin D3,and CDK 2,thus blocking the cell cycle in G0/G1 phase,reducing cell migration,and regulating the PI3K/Akt/mTOR signaling pathway to induce apoptosis.