Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from ...Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from neonatal mouse brain were used.Astrocyte injury was induced via oxygen-glucose deprivation/re-oxygenation(OGD/R).Cell morphology,viability,lactate dehydrogenase(LDH)leakage,apoptosis,glutamate uptake,and brain-derived neurotrophic factor(BDNF)secretion were assessed to gauge cell survival and functionality.Western blot was used to investigate the cyclic adenosine monophosphate(cAMP)and protein kinase B(Akt)signaling pathways.GPCR-G_(s)-specific inhibitors and molecular docking were used to identify target receptors.Results:Rb1 at concentrations ranging from 0.8 to 5μM did not significantly affect the viability,glutamate uptake,or BDNF secretion in normal astrocytes.OGD/R reduced astrocyte viability,increasing their LDH leakage and apoptosis rate.It also decreased glutamate uptake and BDNF secretion by these cells.Rb1 had protective effects of astrocytes challenged by OGD/R,by improving viability,reducing apoptosis,and enhancing glutamate uptake and BDNF secretion.Additionally,Rb1 activated the cAMP and Akt pathways in these cells.When the GPCR-G_(s) inhibitor NF449 was introduced,the protective effects of Rb1 completely disappeared,and its activation of cAMP and Akt signaling pathways was significantly inhibited.Conclusion:Rb1 protects against astrocytes from OGD/R-induced injury through GPCR-G_(s) mediation.展开更多
Mitochondrial damage caused by oxidative stress and energy deficiency induced by focal ischemia and hypoxia are important factors that aggravate diseases.Studies have shown that ginsenoside Rb1 has neurotrophic and ne...Mitochondrial damage caused by oxidative stress and energy deficiency induced by focal ischemia and hypoxia are important factors that aggravate diseases.Studies have shown that ginsenoside Rb1 has neurotrophic and neuroprotective effects.However,whether it influences energy metabolism after spinal cord injury remains unclear.In this study,we treated mouse and cell models of spinal cord injury with ginsenoside Rb1.We found that ginsenoside Rb1 remarkably inhibited neuronal oxidative stress,protected mitochondria,promoted neuronal metabolic reprogramming,increased glycolytic activity and ATP production,and promoted the survival of motor neurons in the anterior horn and the recovery of motor function in the hind limb.Because sirtuin 3 regulates glycolysis and oxidative stress,mouse and cell models of spinal cord injury were treated with the sirtuin 3 inhibitor 3-TYP.When Sirt3 expression was suppressed,we found that the therapeutic effects of ginsenoside Rb1 on spinal cord injury were remarkably inhibited.Therefore,ginsenoside Rb1 is considered a potential drug for the treatment of spinal cord injury,and its therapeutic effects are closely related to sirtuin 3.展开更多
OBJECTIVE Ginsenoside Rb1(Rb1),an important bioactive ingredient of Panax ginseng,has potent neuroprotective effects.The objective of the study is to elucidate the impact of Rb1 treatment on chronic social defeat stre...OBJECTIVE Ginsenoside Rb1(Rb1),an important bioactive ingredient of Panax ginseng,has potent neuroprotective effects.The objective of the study is to elucidate the impact of Rb1 treatment on chronic social defeat stress(CSDS)-induced depressive-like behaviors and its related mechanism.METHODS AND RE⁃SULTS The daily oral administration of Rb1(35 and 70 mg·kg-1)and imipramine(15 mg·kg-1)for 28 d significantly reversed the social avoidance behavior,anhedonia,and behavioral despair via CSDS exposure,as demonstrated by the consid⁃erable elevation in the time in the zone in social interaction test and consumption of sucrose solu⁃tion in sucrose preference test and decrease of immobility time in forced swim test.Moreover,Rb1 obviously restored the CSDS-induced decrease of BDNF-signaling pathway and hippo⁃campal neurogenesis.Rb1 significantly increased the hippocampal levels of ERK,AKT,and CREB phosphorylation and increased the number of DCX+cells in DG.Importantly,the antidepres⁃sant effects of Rb1 were completely blocked in mice by using K252a(the nonselective tyrosine kinase B inhibitor).CONCLUSION Rb1 exerts promising antidepressant-like effects in mice with CSDS-induced depression,and its effects was facilitated by enhancing the BDNF signaling cas⁃cade and up-regulation of hippocampal neuro⁃genesis.展开更多
OBJECTIVE To evaluate whether ginsenoside Rb1 has antiepileptic effects on pen⁃tylenetetrazole(PTZ)-induced epileptic mice via intranasal therapeutic administration.METHODS Rb1 monoclonal antibody was used to observe ...OBJECTIVE To evaluate whether ginsenoside Rb1 has antiepileptic effects on pen⁃tylenetetrazole(PTZ)-induced epileptic mice via intranasal therapeutic administration.METHODS Rb1 monoclonal antibody was used to observe the distribution of Rb120 mg·kg-1 in mouse brain tissues under different administration routes and to explore the feasibility of intranasal Rb1.PTZ was injected intraperitoneally into healthy ICR mice every 48 hours to construct a tonic-clonic epileptic model.Then Rb120 or 40 mg·kg-1 or valproate 300 mg·kg-1 or saline was administered intranasally for 30 d,and PTZ was continued every five days to imitate occa⁃sional convulsions in the clinic.Racine scale(RCS)and wireless electroencephalogram(EEG)monitoring were used to assess the presence and severity of seizure.Immunofluorescence(IF)was performed after drug treatment to evalu⁃ate the effect of Rb1 on brain neuron,microglia and astrocyte in epileptic mice.RESULTS Rb1 had specific binding with anti-Rb1 in the brain under different administration routes,and intrana⁃sal Rb1 was able to enter the brain and play a therapeutic role(P<0.01).PTZ-injured mice pre⁃sented body mass loss,higher seizure stage and shorter seizure latency.At the same time,epilep⁃tic waves,mainly spikes,were detected by wire⁃less EEG.Compared with PTZ group,intranasal Rb1 increased mice weight(P<0.01)and seizure latency(P<0.05),reduced seizure stage(P<0.01)and EEG spikes.In addition,Rb1 significantly reduced neuron loss(P<0.01)indicated by NeuN staining and decreased the number of acti⁃vated microglia(P<0.01)indicated by Iba-1 staining in the cortex and CA1 area of hippocam⁃pus.Moreover,Rb1 reduced the decrease of GLT-1 and GS expression(P<0.05)induced by PTZ.CONCLUTION Intranasal Rb1 has anti-epi⁃leptic effects on PTZ mice.Moreover,Intranasal Rb1 affects the functions of neurons,astrocytes and microglia through regulating the expression of GLT and GS in astrocytes,which may be related to its anti-epileptic effect.展开更多
OBJECTIVE To investigate the anti-arthritic effect and mechanism of action of ginsenoside Rb1 on adju⁃vant-induced arthritis(AIA)in rats.METHODS Male SD rats were received 0.1 mL injections of FCA(10 g·L^-1)emuls...OBJECTIVE To investigate the anti-arthritic effect and mechanism of action of ginsenoside Rb1 on adju⁃vant-induced arthritis(AIA)in rats.METHODS Male SD rats were received 0.1 mL injections of FCA(10 g·L^-1)emulsion into the right hind metatarsal foot pad for arthritis induction.After that,rats were randomly divided into six groups,namely control group,untreated group,dexamethasone(DEX,2.5 mg·kg^-1)group,low(5 mg·kg^-1),medium(10 mg·kg^-1)and high(20 mg·kg^-1)doses of ginsenoside Rb1 groups,and treated intraperitoneally at the above dosage once a day for 2 weeks.After treatment,paw swelling and arthritis indexes were evaluated,the thymus and spleen index were calculated as well.HE staining were used to observe the joint histopathology in rats.Rat ELISA kits were used to determinate the TNF-α,IL-1βand IL-6 levels.Western blotting were used to detect the related protein expression of NF-κB signaling pathway in the tissues of inflamed joints.RESULTS Rb1 significantly decreased the paw swelling and arthritis index,Compared with AIA group.HE staining results revealed that medium and high doses of Rb1 significantly reduced synovial inflammatory cell infiltration,synovial lining hyperplasia and bone destruction,compared with AIA group.Elisa results showed that Rb1 significantly decreased the TNF-α,IL-1β and IL-6 levels(P<0.05,P<0.01).Western blotting results revealed that the expression of p-IκB and p-P65 were significantly reduced in 20 mg·kg^-1 of Rb1 group,compared with AIA group(P<0.05,P<0.01).CONCIUSION Rb1 manifests therapeutic anti-inflammatory effects on rats with AIA,poten⁃tially through a mechanism of inhibiting activation of the NF-κB.展开更多
Objective:To evaluate whether ginsenoside Rb1(Rb1) can attenuate lipopolysaccharide(LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.Methods:Intraperitoneal i...Objective:To evaluate whether ginsenoside Rb1(Rb1) can attenuate lipopolysaccharide(LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.Methods:Intraperitoneal injection with an escalating dose of LPS was used to establish a chronic neuroinflammation model in mice.Once LPS was initiated,10 or 20 mg/kg Rbl,or sterile saline,was administered for 14 consecutive days.Open field test and beam walking test were used to monitor the changes in behavior.The concentration of cytokines in the serum and brain were used to monitor the systemic inflammation and neuroinflammation,respectively.Molecules specific to each glial cell phenotype were used to investigate glial cell polarization.Results:Mice in the LPS group had reduced spontaneous activities and impaired beam walking performance.Rbl obviously eased LPS-induced behavior distu rbances.Regarding the levels of serum cytokines,both tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were significantly increased,while interleukin-10(IL-10) and transforming growth factor β(TGF-β) remarkably decreased after LPS treatment(all P <.001).Rb1 treatment significantly attenuated LPS-induced serum cytokine changes(all P <.05).The results of quantitative polymerase chain reaction and western blotting showed that the mRNA and protein expression levels of TNF-α and complement component 3(C3) in the brain were significantly increased after LPS treatment(all P<.01).Rbl treatment significantly inhibited LPS-induced inflammation in the brain(all P <.05).Glial cell polarization analysis showed that M1 and M2 microglia,and A1 astrocytes increased following LPS treatment,while A2 astrocytes decreased.Rb1 treatment reduced M1 and M2 microglia,and A1 astrocytes,and significantly increased A2 astrocytes.Conclusion:Rb1 can attenuate chronic neuroinflammation induced by LPS in mice,which may be partially attributable to its fine tuning of microglia and astrocyte polarization.Rb1 has potential value for treating neurodegenerative diseases.展开更多
Objective:To investigate whether ginsenoside Rb1(Rb1)can protect human umbilical vein endothelial cells(HUVECs)against high glucose-induced apoptosis and examine the underlying mechanism.Methods:HUVECs were divided in...Objective:To investigate whether ginsenoside Rb1(Rb1)can protect human umbilical vein endothelial cells(HUVECs)against high glucose-induced apoptosis and examine the underlying mechanism.Methods:HUVECs were divided into 5 groups:control group(5.5 mmol/L glucose),high glucose(HG,40 mmol/L)treatment group,Rb1(50μmol/L)treatment group,Rb1 plus HG treatment group,and Rb1 and 3-(1 H-1,2,3-triazol-4-yl)pyridine(3-TYP,16μmol/L)plus HG treatment group.Cell viability was evaluated by cell counting kit-8 assay.Mitochondrial and intracellular reactive oxygen species were detected by Mito Sox Red mitochondrial superoxide indicator and dichloro-dihydro-fluorescein diacetate assay,respectively.Annexin V/propidium iodide staining and fluorescent dye staining were used to measure the apoptosis and the mitochondrial membrane potential of HUVECs,respectively.The protein expressions of apoptosis-related proteins[Bcl-2,Bax,cleaved caspase-3 and cytochrome c(Cyt-c)],mitochondrial biogenesis-related proteins[proliferator-activated receptor gamma coactivator 1-alpha,nuclear respiratory factor-1 and mitochondrial transcription factor A],acetylation levels of forkhead box O3 a and SOD2,and sirtuin-3(SIRT3)signalling pathway were measured by immunoblotting and immunoprecipitation.Results:Rb1 ameliorated survival in cells in which apoptosis was induced by high glucose(P<0.05 or P<0.01).Upon the addition of Rb1,mitochondrial and intracellular reactive oxygen species generation and malondialdehyde levels were decreased(P<0.01),while the activities of antioxidant enzymes were increased(P<0.05 or P<0.01).Rb1 preserved the mitochondrial membrane potential and reduced the release of Cyt-c from the mitochondria into the cytosol(P<0.01).In addition,Rb1 upregulated mitochondrial biogenesis-associated proteins(P<0.01).Notably,the cytoprotective effects of Rb1 were correlated with SIRT3 signalling pathway activation(P<0.01).The effect of Rb1 against high glucose-induced mitochondria-related apoptosis was restrained by 3-TYP(P<0.05 or P<0.01).Conclusion:Rb1 could protect HUVECs from high glucose-induced apoptosis by promoting mitochondrial function and suppressing oxidative stress through the SIRT3 signalling pathway.展开更多
Ischemic stroke causes brain inflammation and multi-organ injury,which is closely associated with the peroxisome proliferator-activated receptor-gamma(PPARγ)signaling pathway.Recent studies have indicated that ginsen...Ischemic stroke causes brain inflammation and multi-organ injury,which is closely associated with the peroxisome proliferator-activated receptor-gamma(PPARγ)signaling pathway.Recent studies have indicated that ginsenoside Rb1(GRb1)can protect the integrity of the blood-brain barrier after stroke.In the current study,a mouse model of middle cerebral artery occlusion/reperfusion(MCAO/R)was established to determine whether GRb1 can ameliorate brain/lung/intestinal barrier damage via the PPARγsignaling pathway.Staining(2,3,5-triphenyltetrazolium chloride,hematoxylin,and eosin)and Doppler ultrasonography were employed to detect pathological changes.Endothelial breakdown was investigated with the leakage of Evans Blue dye and the expression of TJs(tight junctions)and AJs(adherent junctions).Western blot and immunofluorescence were used to determine the levels of cell junction proteins,PPARγand NF-κB.Results showed that GRb1 significantly mitigated multi-organ injury and increased the expression of cerebral microvascular,pulmonary vascular,and intestinal epithelial connexins.In brain,lung,and intestinal tissues,GRb1 activated PPARγ,decreased the levels of phospho-NF-κB p65,and inhibited the production of proinflammatory cytokines,thereby maintaining barrier permeability.However,co-treatment with GRb1 and the PPARγantagonist GW9662 reversed the barrier-protective effect of GRb1.These findings indicated that GRb1 can improve stroke-induced brain/lung/intestinal barrier damagevia the PPARγpathway.展开更多
Objective:The aim of this study was to investigate the protective effects of ginsenoside Rb1 and assess whether these protective effects are related to calcium/calmodulin-dependent protein kinaseⅡ(Ca MKⅡ).Methods:A ...Objective:The aim of this study was to investigate the protective effects of ginsenoside Rb1 and assess whether these protective effects are related to calcium/calmodulin-dependent protein kinaseⅡ(Ca MKⅡ).Methods:A myocardial ischemia(IS)rat.model and a myocardial H9 C2 cell hypoxia model were established.MI was induced by occluding the left anterior descending artery for 120 min.Ginsenoside Rb1(10 mg/kg)was administered 30 min before ischemia induction,and the treatment continued for 7 days.Results:In the rat IS injury model,ginsenoside Rb1 reduced myocardial infarct size,mean left ventricular diastolic pressure,incidence of arrhythmia,and levels of serum creatine kinase,lactate dehydrogenase,and malondialdehyde.However,the mean left ventricular systolic pressure,and maximal rising and falling rates of ventricular pressure(±dp/dtmax)increased.In the myocardial H9 C2 cell hypoxia model,ginsenoside Rb1 reduced intracellular calcium concentrations([Ca2+]i)during hypoxia,and markedly reversed the hypoxia-induced decrease in cell survival.Ginsenoside Rb1 was involved in the downregulation of CaMKⅡand the ryanodine receptor,as well as hypoxia-induced H9 C2 cell survival.Conclusion:The findings of the present study suggest that ginsenoside Rb1 attenuates MI injury in rats,partially through the downregulation of CaMKⅡexpression.展开更多
Ginsenoside Rb1 shows a strong antioxidant effect and has potential activation effects on Akt.The aim of the present study was to investigate the protective effect of Rb1 on age-related ovarian granulosa cell injury.O...Ginsenoside Rb1 shows a strong antioxidant effect and has potential activation effects on Akt.The aim of the present study was to investigate the protective effect of Rb1 on age-related ovarian granulosa cell injury.Ovarian granulosa cells(GCs)were obtained from 50 young women(≤30 years)and 50 aged women(≥38 years)at an IVF center.Young and aged ICR mice were administered with or without Rb1(10 mg kg^(-1),i.p.)for 2 weeks.The protective effects of Rb1 were investigated and the role of Rb1 on the modulation of Akt-FoxO1 interaction was determined with immunofluorescence,Western blotting,immunoprecipitation,si RNA silencing and pharmacological inhibitor.Rb1 effectively decreased LDH and MDA,and reversed the apoptotic-related protein levels in h GL cells from old patients.Similar results were found in mice.In addition,the mitochondrial membrane potential was restored and the overaccumulation of ROS was reversed by Rb1.Rb1 preserved peroxide-impaired Akt activation,to some extent,by increasing phosphorylation at Ser473.Rb1 also facilitated p-Akt binding to FoxO1 and promoted the phosphorylation of FoxO1.Si RNA silencing of Akt,Akt inhibitor LY294002,and FoxO1 inhibitor AS1842856 attenuated the effects of Rb1.Ginsenoside Rb1 inhibits age-related GCs oxidative damage by activating Akt phosphorylation at Ser473 and by further interaction with FoxO1.展开更多
Background Myocardial ischemia/reperfusion injury(MI/RI)during myocardial infarction worsens outcomes. It has been proved that ginsenoside Rb1 has a great impact on ischemia/reperfusion(I/R)injury. The aim of this stu...Background Myocardial ischemia/reperfusion injury(MI/RI)during myocardial infarction worsens outcomes. It has been proved that ginsenoside Rb1 has a great impact on ischemia/reperfusion(I/R)injury. The aim of this study is to explore the protective effect of the pretreatment with ginsenoside Rb1 on MI/RI and investigate the underlying mechanisms about the preventive action. Methods A total of 27 healthy adult Sprague-Dawley(SD)rats were randomly divided into 3 groups(n=9 per group):A sham-operated group(Sham n=9);an ischemia 40 min/reperfusion 2 h(I/R n=9)of the cardiac muscle group;Rb1-treated group was divided into 3 subgroups(Rb1 10 mg/kg,20 mg/kg,40 mg/kg,n=3). Ginsenosides Rb1 at different concentrations were injected intraperitoneally for 3 days continuously before ligation. A model of MI/RI was constructed by ligation of the left coronary artery anterior descending branch in rats. Myocardial infarction area after I/R was measured bytriphenyltetrazolium chloride(TTC)staining of myocardial tissue. Hematoxylin and eosin(HE)staining and electrocardiogram indication were used to observe myocardial cell injury and ischemia,respectively. The expression of caspase-8 protein was observed by immunohistochemistry staining. Results The pretreatment of 40 mg/kg dose of ginsenoside Rb1 could decrease the expression of caspase-8 protein caused by I/R and the apoptosis of myocardial cells,improve myocardial ischemia,reduce the area of myocardial infarction and ameliorate MI/RI.Conclusions These results demonstrate that ginsenoside Rb1 has a significant protective effect on MI/RI through attenuating the apoptosis of myocardial cells and improving myocardial ischemia at appropriate dose,which provides new insights into the potential therapy of MI/RI.展开更多
基金supported by the grant International Cooperation Project of Prevention and Treatment of Major Diseases with Chinese Medicine(GZYYGJ2021047)the High-end Experts Support Program from the Ministry of Science and Technology(DL 2021110001L)the Basic Research Funds from the Ministry of Education(1000061223731).
文摘Objectives:To investigate whether the protective actions of ginsenoside Rb1(Rb1)on astrocytes are mediated through the G_(s)-type G-protein-coupled receptor(GPCR-G_(s)).Methods:Primary astrocyte cultures derived from neonatal mouse brain were used.Astrocyte injury was induced via oxygen-glucose deprivation/re-oxygenation(OGD/R).Cell morphology,viability,lactate dehydrogenase(LDH)leakage,apoptosis,glutamate uptake,and brain-derived neurotrophic factor(BDNF)secretion were assessed to gauge cell survival and functionality.Western blot was used to investigate the cyclic adenosine monophosphate(cAMP)and protein kinase B(Akt)signaling pathways.GPCR-G_(s)-specific inhibitors and molecular docking were used to identify target receptors.Results:Rb1 at concentrations ranging from 0.8 to 5μM did not significantly affect the viability,glutamate uptake,or BDNF secretion in normal astrocytes.OGD/R reduced astrocyte viability,increasing their LDH leakage and apoptosis rate.It also decreased glutamate uptake and BDNF secretion by these cells.Rb1 had protective effects of astrocytes challenged by OGD/R,by improving viability,reducing apoptosis,and enhancing glutamate uptake and BDNF secretion.Additionally,Rb1 activated the cAMP and Akt pathways in these cells.When the GPCR-G_(s) inhibitor NF449 was introduced,the protective effects of Rb1 completely disappeared,and its activation of cAMP and Akt signaling pathways was significantly inhibited.Conclusion:Rb1 protects against astrocytes from OGD/R-induced injury through GPCR-G_(s) mediation.
基金supported by the National Natural Science Foundation of ChinaNos.81871556+2 种基金82072165Liaoning Revitalization Talents ProgramNo.XLYC1902108 (all to XFM)
文摘Mitochondrial damage caused by oxidative stress and energy deficiency induced by focal ischemia and hypoxia are important factors that aggravate diseases.Studies have shown that ginsenoside Rb1 has neurotrophic and neuroprotective effects.However,whether it influences energy metabolism after spinal cord injury remains unclear.In this study,we treated mouse and cell models of spinal cord injury with ginsenoside Rb1.We found that ginsenoside Rb1 remarkably inhibited neuronal oxidative stress,protected mitochondria,promoted neuronal metabolic reprogramming,increased glycolytic activity and ATP production,and promoted the survival of motor neurons in the anterior horn and the recovery of motor function in the hind limb.Because sirtuin 3 regulates glycolysis and oxidative stress,mouse and cell models of spinal cord injury were treated with the sirtuin 3 inhibitor 3-TYP.When Sirt3 expression was suppressed,we found that the therapeutic effects of ginsenoside Rb1 on spinal cord injury were remarkably inhibited.Therefore,ginsenoside Rb1 is considered a potential drug for the treatment of spinal cord injury,and its therapeutic effects are closely related to sirtuin 3.
基金Ministry of Science and Tech⁃nology of China(2017ZX09301029)and Space Medical Experiment Project of China Manned Space Program(HYZHXM05003)。
文摘OBJECTIVE Ginsenoside Rb1(Rb1),an important bioactive ingredient of Panax ginseng,has potent neuroprotective effects.The objective of the study is to elucidate the impact of Rb1 treatment on chronic social defeat stress(CSDS)-induced depressive-like behaviors and its related mechanism.METHODS AND RE⁃SULTS The daily oral administration of Rb1(35 and 70 mg·kg-1)and imipramine(15 mg·kg-1)for 28 d significantly reversed the social avoidance behavior,anhedonia,and behavioral despair via CSDS exposure,as demonstrated by the consid⁃erable elevation in the time in the zone in social interaction test and consumption of sucrose solu⁃tion in sucrose preference test and decrease of immobility time in forced swim test.Moreover,Rb1 obviously restored the CSDS-induced decrease of BDNF-signaling pathway and hippo⁃campal neurogenesis.Rb1 significantly increased the hippocampal levels of ERK,AKT,and CREB phosphorylation and increased the number of DCX+cells in DG.Importantly,the antidepres⁃sant effects of Rb1 were completely blocked in mice by using K252a(the nonselective tyrosine kinase B inhibitor).CONCLUSION Rb1 exerts promising antidepressant-like effects in mice with CSDS-induced depression,and its effects was facilitated by enhancing the BDNF signaling cas⁃cade and up-regulation of hippocampal neuro⁃genesis.
文摘OBJECTIVE To evaluate whether ginsenoside Rb1 has antiepileptic effects on pen⁃tylenetetrazole(PTZ)-induced epileptic mice via intranasal therapeutic administration.METHODS Rb1 monoclonal antibody was used to observe the distribution of Rb120 mg·kg-1 in mouse brain tissues under different administration routes and to explore the feasibility of intranasal Rb1.PTZ was injected intraperitoneally into healthy ICR mice every 48 hours to construct a tonic-clonic epileptic model.Then Rb120 or 40 mg·kg-1 or valproate 300 mg·kg-1 or saline was administered intranasally for 30 d,and PTZ was continued every five days to imitate occa⁃sional convulsions in the clinic.Racine scale(RCS)and wireless electroencephalogram(EEG)monitoring were used to assess the presence and severity of seizure.Immunofluorescence(IF)was performed after drug treatment to evalu⁃ate the effect of Rb1 on brain neuron,microglia and astrocyte in epileptic mice.RESULTS Rb1 had specific binding with anti-Rb1 in the brain under different administration routes,and intrana⁃sal Rb1 was able to enter the brain and play a therapeutic role(P<0.01).PTZ-injured mice pre⁃sented body mass loss,higher seizure stage and shorter seizure latency.At the same time,epilep⁃tic waves,mainly spikes,were detected by wire⁃less EEG.Compared with PTZ group,intranasal Rb1 increased mice weight(P<0.01)and seizure latency(P<0.05),reduced seizure stage(P<0.01)and EEG spikes.In addition,Rb1 significantly reduced neuron loss(P<0.01)indicated by NeuN staining and decreased the number of acti⁃vated microglia(P<0.01)indicated by Iba-1 staining in the cortex and CA1 area of hippocam⁃pus.Moreover,Rb1 reduced the decrease of GLT-1 and GS expression(P<0.05)induced by PTZ.CONCLUTION Intranasal Rb1 has anti-epi⁃leptic effects on PTZ mice.Moreover,Intranasal Rb1 affects the functions of neurons,astrocytes and microglia through regulating the expression of GLT and GS in astrocytes,which may be related to its anti-epileptic effect.
文摘OBJECTIVE To investigate the anti-arthritic effect and mechanism of action of ginsenoside Rb1 on adju⁃vant-induced arthritis(AIA)in rats.METHODS Male SD rats were received 0.1 mL injections of FCA(10 g·L^-1)emulsion into the right hind metatarsal foot pad for arthritis induction.After that,rats were randomly divided into six groups,namely control group,untreated group,dexamethasone(DEX,2.5 mg·kg^-1)group,low(5 mg·kg^-1),medium(10 mg·kg^-1)and high(20 mg·kg^-1)doses of ginsenoside Rb1 groups,and treated intraperitoneally at the above dosage once a day for 2 weeks.After treatment,paw swelling and arthritis indexes were evaluated,the thymus and spleen index were calculated as well.HE staining were used to observe the joint histopathology in rats.Rat ELISA kits were used to determinate the TNF-α,IL-1βand IL-6 levels.Western blotting were used to detect the related protein expression of NF-κB signaling pathway in the tissues of inflamed joints.RESULTS Rb1 significantly decreased the paw swelling and arthritis index,Compared with AIA group.HE staining results revealed that medium and high doses of Rb1 significantly reduced synovial inflammatory cell infiltration,synovial lining hyperplasia and bone destruction,compared with AIA group.Elisa results showed that Rb1 significantly decreased the TNF-α,IL-1β and IL-6 levels(P<0.05,P<0.01).Western blotting results revealed that the expression of p-IκB and p-P65 were significantly reduced in 20 mg·kg^-1 of Rb1 group,compared with AIA group(P<0.05,P<0.01).CONCIUSION Rb1 manifests therapeutic anti-inflammatory effects on rats with AIA,poten⁃tially through a mechanism of inhibiting activation of the NF-κB.
基金This research was supported by the grant“Prevention and Treatment of Major Diseases with Chinese Medicine from the National Administration of Traditional Chinese Medicine(GZYYGJ2021047)”the“High-end Experts Support Program from the Ministry of Science and Technology(DL 2021110001L)”the“Basic Research Funds from the Ministry of Education 1000061223731)”。
文摘Objective:To evaluate whether ginsenoside Rb1(Rb1) can attenuate lipopolysaccharide(LPS)-induced chronic neuroinflammation in mice and to explore its relationship with glial cell polarization.Methods:Intraperitoneal injection with an escalating dose of LPS was used to establish a chronic neuroinflammation model in mice.Once LPS was initiated,10 or 20 mg/kg Rbl,or sterile saline,was administered for 14 consecutive days.Open field test and beam walking test were used to monitor the changes in behavior.The concentration of cytokines in the serum and brain were used to monitor the systemic inflammation and neuroinflammation,respectively.Molecules specific to each glial cell phenotype were used to investigate glial cell polarization.Results:Mice in the LPS group had reduced spontaneous activities and impaired beam walking performance.Rbl obviously eased LPS-induced behavior distu rbances.Regarding the levels of serum cytokines,both tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) were significantly increased,while interleukin-10(IL-10) and transforming growth factor β(TGF-β) remarkably decreased after LPS treatment(all P <.001).Rb1 treatment significantly attenuated LPS-induced serum cytokine changes(all P <.05).The results of quantitative polymerase chain reaction and western blotting showed that the mRNA and protein expression levels of TNF-α and complement component 3(C3) in the brain were significantly increased after LPS treatment(all P<.01).Rbl treatment significantly inhibited LPS-induced inflammation in the brain(all P <.05).Glial cell polarization analysis showed that M1 and M2 microglia,and A1 astrocytes increased following LPS treatment,while A2 astrocytes decreased.Rb1 treatment reduced M1 and M2 microglia,and A1 astrocytes,and significantly increased A2 astrocytes.Conclusion:Rb1 can attenuate chronic neuroinflammation induced by LPS in mice,which may be partially attributable to its fine tuning of microglia and astrocyte polarization.Rb1 has potential value for treating neurodegenerative diseases.
基金Supported by the National Natural Science Foundation of China(No.81370447)Science and Technology Planning Project of Guangdong Province,China(No.2016A050502014)the Ph.D.Start-up Fund of Natural Science Foundation of Guangdong Province,China(No.2015A030310048,and 2016A030310203)。
文摘Objective:To investigate whether ginsenoside Rb1(Rb1)can protect human umbilical vein endothelial cells(HUVECs)against high glucose-induced apoptosis and examine the underlying mechanism.Methods:HUVECs were divided into 5 groups:control group(5.5 mmol/L glucose),high glucose(HG,40 mmol/L)treatment group,Rb1(50μmol/L)treatment group,Rb1 plus HG treatment group,and Rb1 and 3-(1 H-1,2,3-triazol-4-yl)pyridine(3-TYP,16μmol/L)plus HG treatment group.Cell viability was evaluated by cell counting kit-8 assay.Mitochondrial and intracellular reactive oxygen species were detected by Mito Sox Red mitochondrial superoxide indicator and dichloro-dihydro-fluorescein diacetate assay,respectively.Annexin V/propidium iodide staining and fluorescent dye staining were used to measure the apoptosis and the mitochondrial membrane potential of HUVECs,respectively.The protein expressions of apoptosis-related proteins[Bcl-2,Bax,cleaved caspase-3 and cytochrome c(Cyt-c)],mitochondrial biogenesis-related proteins[proliferator-activated receptor gamma coactivator 1-alpha,nuclear respiratory factor-1 and mitochondrial transcription factor A],acetylation levels of forkhead box O3 a and SOD2,and sirtuin-3(SIRT3)signalling pathway were measured by immunoblotting and immunoprecipitation.Results:Rb1 ameliorated survival in cells in which apoptosis was induced by high glucose(P<0.05 or P<0.01).Upon the addition of Rb1,mitochondrial and intracellular reactive oxygen species generation and malondialdehyde levels were decreased(P<0.01),while the activities of antioxidant enzymes were increased(P<0.05 or P<0.01).Rb1 preserved the mitochondrial membrane potential and reduced the release of Cyt-c from the mitochondria into the cytosol(P<0.01).In addition,Rb1 upregulated mitochondrial biogenesis-associated proteins(P<0.01).Notably,the cytoprotective effects of Rb1 were correlated with SIRT3 signalling pathway activation(P<0.01).The effect of Rb1 against high glucose-induced mitochondria-related apoptosis was restrained by 3-TYP(P<0.05 or P<0.01).Conclusion:Rb1 could protect HUVECs from high glucose-induced apoptosis by promoting mitochondrial function and suppressing oxidative stress through the SIRT3 signalling pathway.
基金supported by the National Natural Science Foundation of China(Nos.81773971,82074058,82104438,and 82104437)the China Postdoctoral Science Foundation(Nos.2021M693518 and 2021M693519)+2 种基金the Natural Science Foundation of Jiangsu Province(Nos.SBK20210432 and SBK20210431)the National Science Foundation for the Third Batch of Special Funding for Postdoctoral Fellows(No.2021TQ0367)the“Double First-Class”University Project(No.CPU2018GF07)。
文摘Ischemic stroke causes brain inflammation and multi-organ injury,which is closely associated with the peroxisome proliferator-activated receptor-gamma(PPARγ)signaling pathway.Recent studies have indicated that ginsenoside Rb1(GRb1)can protect the integrity of the blood-brain barrier after stroke.In the current study,a mouse model of middle cerebral artery occlusion/reperfusion(MCAO/R)was established to determine whether GRb1 can ameliorate brain/lung/intestinal barrier damage via the PPARγsignaling pathway.Staining(2,3,5-triphenyltetrazolium chloride,hematoxylin,and eosin)and Doppler ultrasonography were employed to detect pathological changes.Endothelial breakdown was investigated with the leakage of Evans Blue dye and the expression of TJs(tight junctions)and AJs(adherent junctions).Western blot and immunofluorescence were used to determine the levels of cell junction proteins,PPARγand NF-κB.Results showed that GRb1 significantly mitigated multi-organ injury and increased the expression of cerebral microvascular,pulmonary vascular,and intestinal epithelial connexins.In brain,lung,and intestinal tissues,GRb1 activated PPARγ,decreased the levels of phospho-NF-κB p65,and inhibited the production of proinflammatory cytokines,thereby maintaining barrier permeability.However,co-treatment with GRb1 and the PPARγantagonist GW9662 reversed the barrier-protective effect of GRb1.These findings indicated that GRb1 can improve stroke-induced brain/lung/intestinal barrier damagevia the PPARγpathway.
基金supported by the National Natural Science Funds(81073134)085 First-Class Discipline Construction Innovation Science and Technology Support Project of Shanghai University of TCM(085ZY1206)E-institutes of Shanghai Municipal Education Commission(No E 03008)。
文摘Objective:The aim of this study was to investigate the protective effects of ginsenoside Rb1 and assess whether these protective effects are related to calcium/calmodulin-dependent protein kinaseⅡ(Ca MKⅡ).Methods:A myocardial ischemia(IS)rat.model and a myocardial H9 C2 cell hypoxia model were established.MI was induced by occluding the left anterior descending artery for 120 min.Ginsenoside Rb1(10 mg/kg)was administered 30 min before ischemia induction,and the treatment continued for 7 days.Results:In the rat IS injury model,ginsenoside Rb1 reduced myocardial infarct size,mean left ventricular diastolic pressure,incidence of arrhythmia,and levels of serum creatine kinase,lactate dehydrogenase,and malondialdehyde.However,the mean left ventricular systolic pressure,and maximal rising and falling rates of ventricular pressure(±dp/dtmax)increased.In the myocardial H9 C2 cell hypoxia model,ginsenoside Rb1 reduced intracellular calcium concentrations([Ca2+]i)during hypoxia,and markedly reversed the hypoxia-induced decrease in cell survival.Ginsenoside Rb1 was involved in the downregulation of CaMKⅡand the ryanodine receptor,as well as hypoxia-induced H9 C2 cell survival.Conclusion:The findings of the present study suggest that ginsenoside Rb1 attenuates MI injury in rats,partially through the downregulation of CaMKⅡexpression.
基金the National Natural Science Foundation of China(82101714,81925013,81971381,81771580,81771650,82071612,and 81571400)China Postdoctoral Science Foundation(2021M690259,2021M702223)the National Key Research and Development Project of China(2016YFC1000601,2018YFC1004101)。
文摘Ginsenoside Rb1 shows a strong antioxidant effect and has potential activation effects on Akt.The aim of the present study was to investigate the protective effect of Rb1 on age-related ovarian granulosa cell injury.Ovarian granulosa cells(GCs)were obtained from 50 young women(≤30 years)and 50 aged women(≥38 years)at an IVF center.Young and aged ICR mice were administered with or without Rb1(10 mg kg^(-1),i.p.)for 2 weeks.The protective effects of Rb1 were investigated and the role of Rb1 on the modulation of Akt-FoxO1 interaction was determined with immunofluorescence,Western blotting,immunoprecipitation,si RNA silencing and pharmacological inhibitor.Rb1 effectively decreased LDH and MDA,and reversed the apoptotic-related protein levels in h GL cells from old patients.Similar results were found in mice.In addition,the mitochondrial membrane potential was restored and the overaccumulation of ROS was reversed by Rb1.Rb1 preserved peroxide-impaired Akt activation,to some extent,by increasing phosphorylation at Ser473.Rb1 also facilitated p-Akt binding to FoxO1 and promoted the phosphorylation of FoxO1.Si RNA silencing of Akt,Akt inhibitor LY294002,and FoxO1 inhibitor AS1842856 attenuated the effects of Rb1.Ginsenoside Rb1 inhibits age-related GCs oxidative damage by activating Akt phosphorylation at Ser473 and by further interaction with FoxO1.
文摘Background Myocardial ischemia/reperfusion injury(MI/RI)during myocardial infarction worsens outcomes. It has been proved that ginsenoside Rb1 has a great impact on ischemia/reperfusion(I/R)injury. The aim of this study is to explore the protective effect of the pretreatment with ginsenoside Rb1 on MI/RI and investigate the underlying mechanisms about the preventive action. Methods A total of 27 healthy adult Sprague-Dawley(SD)rats were randomly divided into 3 groups(n=9 per group):A sham-operated group(Sham n=9);an ischemia 40 min/reperfusion 2 h(I/R n=9)of the cardiac muscle group;Rb1-treated group was divided into 3 subgroups(Rb1 10 mg/kg,20 mg/kg,40 mg/kg,n=3). Ginsenosides Rb1 at different concentrations were injected intraperitoneally for 3 days continuously before ligation. A model of MI/RI was constructed by ligation of the left coronary artery anterior descending branch in rats. Myocardial infarction area after I/R was measured bytriphenyltetrazolium chloride(TTC)staining of myocardial tissue. Hematoxylin and eosin(HE)staining and electrocardiogram indication were used to observe myocardial cell injury and ischemia,respectively. The expression of caspase-8 protein was observed by immunohistochemistry staining. Results The pretreatment of 40 mg/kg dose of ginsenoside Rb1 could decrease the expression of caspase-8 protein caused by I/R and the apoptosis of myocardial cells,improve myocardial ischemia,reduce the area of myocardial infarction and ameliorate MI/RI.Conclusions These results demonstrate that ginsenoside Rb1 has a significant protective effect on MI/RI through attenuating the apoptosis of myocardial cells and improving myocardial ischemia at appropriate dose,which provides new insights into the potential therapy of MI/RI.
