Epimedium Brevicornum is a traditional Chinese medicinal plant possessing properties of sweet, warm, tonifying kidney, strong bones and rheumatism. Icariin, a flavonoid compound, is one of the main active ingredients ...Epimedium Brevicornum is a traditional Chinese medicinal plant possessing properties of sweet, warm, tonifying kidney, strong bones and rheumatism. Icariin, a flavonoid compound, is one of the main active ingredients of Epimedium. Icariinriside(ICS) is the main metabolite of icariin. Icariinand ICS have multiple pharmacological effects such as anti-tumor, anti-oxidative stress, improvement of cardiovascular and cerebrovascular, and regulation of endocrine. We have conducted a series of studies on the neuroprotection and mechanisms of action of icariin and ICS for many years. The main findings are reported as follows.(1) Effect on Alzheimer disease(AD) model animals: Icariin significantly attenuated learning and memory loss, hippocampal neuron loss and senile plaque formation in APP/PS1 transgenic AD model mice, which may be related to inhibition of Aβ production and reduction of PDE5(phosphodiesterase 5).In addition, icariin significantly attenuated Aβ25-35-induced learning and memory decline and hippocampal neuronal apoptosis in rats, which may be related to lowering PDE5 content and up-regulating BDNF/Trkb/CREB signaling pathway, inhibiting MAPK and NF-κB signaling pathways, and increasing expression of acetylcholinesterase(ACHE) and choline acetyltransferase(CHAT) in the hippocampus. At the same time, icariin can significantly improve the learning and memory dysfunction induced by amanita proline in rats, which may be related to the inhibition of hippocampal neuronal apoptosis, antiexcitatory amino acid toxicity and regulation of MAPK and NF-κB signaling pathways.(2) Effects on Parkinson disease(PD) model animals: The study found that in LPS-induced dopaminergic neuron injury animal models and cell models, icariin can inhibit microglia by inhibiting the expression of inflammatory factors such as TNF-α, IL-1β, NO and COX-2. Activation of glial cells increases the expression of neurotrophic factors such as BDNF and GDNF, increases the content of dopamine(DA) and its metabolites 3, 4-dihydroxyphenylacetic acid(DOPAC) and homovanillic acid(HVA), inhibits MAPK and the NF-κB signaling pathway, protecting dopaminergic neurons. In addition, icariin significantly attenuated6-OHDA-induced dopaminergic neuronal damage. In Nrf2 knockout mice, the neuroprotective effect of icariin disappeared, suggesting that Nrf2 may be one of the targets of icariin to play neuroprotective effects.(3) Effects on vascular dementia(VD) model animals: Icarin can improve the learning and memory ability and memory function of chronic hypoperfusion rats, and its mechanism may be related to increase the level of VEGF/VEGFR2 protein in the brain and activate multiple downstream signaling pathways to promote angiogenesis to play an indirect protective effect on neurons;The level of BDNF/Trk B protein in the brain increases the phosphorylation level of CREB and exerts direct neuroprotective effects.(4)Effect on cerebral ischemia: In a model of ischemic brain injury, icariin acts to up-regulate Sirt1 by activating p38, thereby exerting an anti-ischemic injury and protecting neuronal cells. In addition, icariin has neuroprotective effects on cerebral ischemia-reperfusion injury in rats, which may increase GSH-Px,SOD activity, decrease MDA content, inhibit free radical damage, reduce NO content, NOS activity,and inhibit neurotoxic damage. Reduction of MPO activity, TNF-α, IL-1β content is associated with inhibition of inflammatory response.(5) Cell protection: Icariin has a protective effect on 6-OHDA-induced oxidative damage in PC12 cells, which may be related to inhibition of apoptosis and regulation of Keap1/Nrf2/ARE signaling pathway, while ICS can attenuate oxygen-glucose deprivation/reoxygenation-induced cellular damage in PC12 cells. The mechanism of cellular oxidative damage may be related to inhibition of apoptosis and regulation of Nrf2/SIRT3 signaling pathway.Icariin and ICS have good preventive and therapeutic effects on central nervous system diseases such as AD, PD, VD, etc. However, due to the complexity of the molecular mechanisms of icariin and ICS, the molecular mechanisms of the central nervous system are still worthy of further study.展开更多
OBJECTIVE To investigate the protective effect of icariin(ICA) on learning and memory function in APP/PS1/Tau triple transgenic Alzheimer disease mice(3×Tg-AD mice),and then to explore whether its mechanism is re...OBJECTIVE To investigate the protective effect of icariin(ICA) on learning and memory function in APP/PS1/Tau triple transgenic Alzheimer disease mice(3×Tg-AD mice),and then to explore whether its mechanism is related to the improvement of brain glucose metabolism disorder.METHODS Three-month-old male 3 ×Tg-AD mice were randomly divided into three groups(n=10):3×Tg group,3×Tg+ICA low-dose group(30 mg·kg-1) and 3×Tg + ICA high-dose group(60 mg·kg-1).Age-matched male wild type(WT) mice were randomly divided into two groups(n=10):WT control group and WT+ICA60 mg·kg-1 group.ICA in vehicle(0.5% Tween-80 in distilled water) was given orally once a day for five months in the 3×Tg+ICA groups.3×Tg and WT control group were given an equal volume vehicle.Morris water maze was used to detect the learning and memory function of mice.Brain glucose metabolism in 3×Tg mice was observed by 18 F-FDG microPET imaging technique.Nissl staining and HE staining were used to evaluate the survival neurons in hippocampus of mice.Glucose oxidase assay was used to detect glucose contents in cortex of mice.The protein expression of APP,Aβ1-40,Aβ1-42 and glucose transporter 1(GLUT1),and the phosphorylation level of tau protein at multiple sites in hippocampus were detected by Western blotting.RESULTS Behavioral examination revealed a profound decrease learning and memory function,accompanied by a decrease in number of neuronal cells in 3×Tg-AD mice.Moreover,the cerebral18 F-FDG uptake rate per gram tissue was reduced and the glucose contents in the cortex were increased in 3×Tg-AD mice.In addition,Western blotting analysis showed that the expression of APP,Aβ1-40,Aβ1-42 proteins and the levels of tau protein phosphorylation at Ser199/202 and PHF-1(Ser396/404) sites were increased significantly,followed by a decrease of GLUT1 expression in hippocampus of 3×Tg-AD mice.All of these changes in behavioral functions,neuronal loss and related protein expression were reversed when mice were treated with ICA.CONCLUSION ICA can improve the learning and memory ability of AD model mice,the mechanism may be related to the improvement of cerebral glucose metabolism dysfunction by increasing the expression of GLUT1.展开更多
OBJECTIVE Alzheimer disease(AD) is a progressive neurodegenerative disorder involving a gradual decline in many cognitive processes and in neurons.The endoplasmic reticulum(ER) is involved in several crucial cellular ...OBJECTIVE Alzheimer disease(AD) is a progressive neurodegenerative disorder involving a gradual decline in many cognitive processes and in neurons.The endoplasmic reticulum(ER) is involved in several crucial cellular functions,eg protein folding and quality control.Massive misfolded or unfolded proteins in ER can disturb the function of ER and induce ER stress,which results in neuronal death in AD.Icariin(ICA) has a wide range of neuro protection and has been researched in AD treatment.However,whether ICA has the effect on ER stress in AD condition,and how ICA affects ER stress remains stil unclear.Therefore,the current study aimed to investigate the mechanism of ICA against cognitive impairments in AD model through ER stress pathway and apoptosis.METHODS Twelve months male APP/PS1 or wild-type(WT)mice were randomly divided into four groups:APP/PS1,and APP/PS1+ICA,WT and WT+ICA groups.The treated mice were given ICA60 mg·kg-1 per day and control mice were received the same volume distilled water for consecutive 3 months.The Morris water maze and novel object recognition were used to detect animals′ behavior.Nissl staining was used to observe the neuronal morphology in hippocampus area.The protein and(or) phosphorylation level of GRP78,p-PERK,PERK,p-IRE1,IRE1,ATF6,p-e IF2α,eIF2α,ATF4,CHOP,the level of cleaved-casepase 3,Bax and Bcl-2 were examined by Western blotting.RESULTS The behavior performance testing by Morris water maze and novel object recognition deteriorated in APP/PS1 mice compared with WT mice,however,ICA significantly improved the behavior performance when compared with APP/PS1.The neuron impairments in APP/PS1 mice also were ameliorated after ICA treatment.The protein expression of GRP78,ATF4,CHOP,and the level of p-PERK and p-eI F2α were higher in APP/PS1 mice than that in WT mice.The Bax/Bcl-2 ratio elevation and caspase 3 activation have been found in APP/PS1 mice.After treated with ICA,those above-mentioned parameters were decreased compared with APP/PS1.However,the levels of IRE1,p-IRE1 and ATF6 were not change among other groups.CONCLUSION ICA may decrease the ER stress and apoptosis in AD model,and may be through inhibiting the PERK/eI F2α pathway,not IRE and ATF6 pathways.展开更多
OBJECTIVE To investigate the protective effect of icariin(ICA) on lipopolysaccharide(LPS)-induced BV2 microglia injury,and to clarify the role of nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathway in ...OBJECTIVE To investigate the protective effect of icariin(ICA) on lipopolysaccharide(LPS)-induced BV2 microglia injury,and to clarify the role of nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathway in BV2 microglia-mediated neuroinflammation.METHODS BV2 microglia were randomly divided into control,ICA(0.1 μmol·L^(-1)),LPS(1 mg·L^(-1)),LPS + ICA(0.01 μmol·L^(-1)),and LPS + ICA(0.1 μmol·L^(-1))groups.BV2 microglia were treated with ICA for 30 min and then treated with LPS for 24 h.MTT assay was used to determine the cells survival rate,Griess kit and ELISA kits were used to detect the contents of NO,IL-1β and IL-18 in the culture supernatant,Western blotting was used to detect the expression of Nrf2,HO-1 and NQO1.Real time RT-PCR was used to detect the expression of Nrf2,HO-1 and NQO1 after ICA addition for 2,6 and 24 h.And immunofluorescence was used to observe the activation of Nrf2.RESULTS ICA reduced LPS-induced NO,IL-1β and IL-18 production in the culture supernatant,and ICA increase LPS-induced mRNA and protein expression of Nrf2 signaling pathway.CONCLUSION ICA protects LPS induced neuroinflammation by regulating Nrf2 signaling pathway.展开更多
OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment t...OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.展开更多
文摘Epimedium Brevicornum is a traditional Chinese medicinal plant possessing properties of sweet, warm, tonifying kidney, strong bones and rheumatism. Icariin, a flavonoid compound, is one of the main active ingredients of Epimedium. Icariinriside(ICS) is the main metabolite of icariin. Icariinand ICS have multiple pharmacological effects such as anti-tumor, anti-oxidative stress, improvement of cardiovascular and cerebrovascular, and regulation of endocrine. We have conducted a series of studies on the neuroprotection and mechanisms of action of icariin and ICS for many years. The main findings are reported as follows.(1) Effect on Alzheimer disease(AD) model animals: Icariin significantly attenuated learning and memory loss, hippocampal neuron loss and senile plaque formation in APP/PS1 transgenic AD model mice, which may be related to inhibition of Aβ production and reduction of PDE5(phosphodiesterase 5).In addition, icariin significantly attenuated Aβ25-35-induced learning and memory decline and hippocampal neuronal apoptosis in rats, which may be related to lowering PDE5 content and up-regulating BDNF/Trkb/CREB signaling pathway, inhibiting MAPK and NF-κB signaling pathways, and increasing expression of acetylcholinesterase(ACHE) and choline acetyltransferase(CHAT) in the hippocampus. At the same time, icariin can significantly improve the learning and memory dysfunction induced by amanita proline in rats, which may be related to the inhibition of hippocampal neuronal apoptosis, antiexcitatory amino acid toxicity and regulation of MAPK and NF-κB signaling pathways.(2) Effects on Parkinson disease(PD) model animals: The study found that in LPS-induced dopaminergic neuron injury animal models and cell models, icariin can inhibit microglia by inhibiting the expression of inflammatory factors such as TNF-α, IL-1β, NO and COX-2. Activation of glial cells increases the expression of neurotrophic factors such as BDNF and GDNF, increases the content of dopamine(DA) and its metabolites 3, 4-dihydroxyphenylacetic acid(DOPAC) and homovanillic acid(HVA), inhibits MAPK and the NF-κB signaling pathway, protecting dopaminergic neurons. In addition, icariin significantly attenuated6-OHDA-induced dopaminergic neuronal damage. In Nrf2 knockout mice, the neuroprotective effect of icariin disappeared, suggesting that Nrf2 may be one of the targets of icariin to play neuroprotective effects.(3) Effects on vascular dementia(VD) model animals: Icarin can improve the learning and memory ability and memory function of chronic hypoperfusion rats, and its mechanism may be related to increase the level of VEGF/VEGFR2 protein in the brain and activate multiple downstream signaling pathways to promote angiogenesis to play an indirect protective effect on neurons;The level of BDNF/Trk B protein in the brain increases the phosphorylation level of CREB and exerts direct neuroprotective effects.(4)Effect on cerebral ischemia: In a model of ischemic brain injury, icariin acts to up-regulate Sirt1 by activating p38, thereby exerting an anti-ischemic injury and protecting neuronal cells. In addition, icariin has neuroprotective effects on cerebral ischemia-reperfusion injury in rats, which may increase GSH-Px,SOD activity, decrease MDA content, inhibit free radical damage, reduce NO content, NOS activity,and inhibit neurotoxic damage. Reduction of MPO activity, TNF-α, IL-1β content is associated with inhibition of inflammatory response.(5) Cell protection: Icariin has a protective effect on 6-OHDA-induced oxidative damage in PC12 cells, which may be related to inhibition of apoptosis and regulation of Keap1/Nrf2/ARE signaling pathway, while ICS can attenuate oxygen-glucose deprivation/reoxygenation-induced cellular damage in PC12 cells. The mechanism of cellular oxidative damage may be related to inhibition of apoptosis and regulation of Nrf2/SIRT3 signaling pathway.Icariin and ICS have good preventive and therapeutic effects on central nervous system diseases such as AD, PD, VD, etc. However, due to the complexity of the molecular mechanisms of icariin and ICS, the molecular mechanisms of the central nervous system are still worthy of further study.
基金National Natural Science Foundation of China(81660599)Foundation of Zunyi Medical University (2013F-686+1 种基金2013F-738)Postgraduate Education Foundation of Guizhou Province(KYJJ2017008).
文摘OBJECTIVE To investigate the protective effect of icariin(ICA) on learning and memory function in APP/PS1/Tau triple transgenic Alzheimer disease mice(3×Tg-AD mice),and then to explore whether its mechanism is related to the improvement of brain glucose metabolism disorder.METHODS Three-month-old male 3 ×Tg-AD mice were randomly divided into three groups(n=10):3×Tg group,3×Tg+ICA low-dose group(30 mg·kg-1) and 3×Tg + ICA high-dose group(60 mg·kg-1).Age-matched male wild type(WT) mice were randomly divided into two groups(n=10):WT control group and WT+ICA60 mg·kg-1 group.ICA in vehicle(0.5% Tween-80 in distilled water) was given orally once a day for five months in the 3×Tg+ICA groups.3×Tg and WT control group were given an equal volume vehicle.Morris water maze was used to detect the learning and memory function of mice.Brain glucose metabolism in 3×Tg mice was observed by 18 F-FDG microPET imaging technique.Nissl staining and HE staining were used to evaluate the survival neurons in hippocampus of mice.Glucose oxidase assay was used to detect glucose contents in cortex of mice.The protein expression of APP,Aβ1-40,Aβ1-42 and glucose transporter 1(GLUT1),and the phosphorylation level of tau protein at multiple sites in hippocampus were detected by Western blotting.RESULTS Behavioral examination revealed a profound decrease learning and memory function,accompanied by a decrease in number of neuronal cells in 3×Tg-AD mice.Moreover,the cerebral18 F-FDG uptake rate per gram tissue was reduced and the glucose contents in the cortex were increased in 3×Tg-AD mice.In addition,Western blotting analysis showed that the expression of APP,Aβ1-40,Aβ1-42 proteins and the levels of tau protein phosphorylation at Ser199/202 and PHF-1(Ser396/404) sites were increased significantly,followed by a decrease of GLUT1 expression in hippocampus of 3×Tg-AD mice.All of these changes in behavioral functions,neuronal loss and related protein expression were reversed when mice were treated with ICA.CONCLUSION ICA can improve the learning and memory ability of AD model mice,the mechanism may be related to the improvement of cerebral glucose metabolism dysfunction by increasing the expression of GLUT1.
基金National Natural Science Foundation of China(81560594).
文摘OBJECTIVE Alzheimer disease(AD) is a progressive neurodegenerative disorder involving a gradual decline in many cognitive processes and in neurons.The endoplasmic reticulum(ER) is involved in several crucial cellular functions,eg protein folding and quality control.Massive misfolded or unfolded proteins in ER can disturb the function of ER and induce ER stress,which results in neuronal death in AD.Icariin(ICA) has a wide range of neuro protection and has been researched in AD treatment.However,whether ICA has the effect on ER stress in AD condition,and how ICA affects ER stress remains stil unclear.Therefore,the current study aimed to investigate the mechanism of ICA against cognitive impairments in AD model through ER stress pathway and apoptosis.METHODS Twelve months male APP/PS1 or wild-type(WT)mice were randomly divided into four groups:APP/PS1,and APP/PS1+ICA,WT and WT+ICA groups.The treated mice were given ICA60 mg·kg-1 per day and control mice were received the same volume distilled water for consecutive 3 months.The Morris water maze and novel object recognition were used to detect animals′ behavior.Nissl staining was used to observe the neuronal morphology in hippocampus area.The protein and(or) phosphorylation level of GRP78,p-PERK,PERK,p-IRE1,IRE1,ATF6,p-e IF2α,eIF2α,ATF4,CHOP,the level of cleaved-casepase 3,Bax and Bcl-2 were examined by Western blotting.RESULTS The behavior performance testing by Morris water maze and novel object recognition deteriorated in APP/PS1 mice compared with WT mice,however,ICA significantly improved the behavior performance when compared with APP/PS1.The neuron impairments in APP/PS1 mice also were ameliorated after ICA treatment.The protein expression of GRP78,ATF4,CHOP,and the level of p-PERK and p-eI F2α were higher in APP/PS1 mice than that in WT mice.The Bax/Bcl-2 ratio elevation and caspase 3 activation have been found in APP/PS1 mice.After treated with ICA,those above-mentioned parameters were decreased compared with APP/PS1.However,the levels of IRE1,p-IRE1 and ATF6 were not change among other groups.CONCLUSION ICA may decrease the ER stress and apoptosis in AD model,and may be through inhibiting the PERK/eI F2α pathway,not IRE and ATF6 pathways.
基金National Natural Science Foundation of China(81760658).
文摘OBJECTIVE To investigate the protective effect of icariin(ICA) on lipopolysaccharide(LPS)-induced BV2 microglia injury,and to clarify the role of nuclear factor erythroid 2-related factor 2(Nrf2) signaling pathway in BV2 microglia-mediated neuroinflammation.METHODS BV2 microglia were randomly divided into control,ICA(0.1 μmol·L^(-1)),LPS(1 mg·L^(-1)),LPS + ICA(0.01 μmol·L^(-1)),and LPS + ICA(0.1 μmol·L^(-1))groups.BV2 microglia were treated with ICA for 30 min and then treated with LPS for 24 h.MTT assay was used to determine the cells survival rate,Griess kit and ELISA kits were used to detect the contents of NO,IL-1β and IL-18 in the culture supernatant,Western blotting was used to detect the expression of Nrf2,HO-1 and NQO1.Real time RT-PCR was used to detect the expression of Nrf2,HO-1 and NQO1 after ICA addition for 2,6 and 24 h.And immunofluorescence was used to observe the activation of Nrf2.RESULTS ICA reduced LPS-induced NO,IL-1β and IL-18 production in the culture supernatant,and ICA increase LPS-induced mRNA and protein expression of Nrf2 signaling pathway.CONCLUSION ICA protects LPS induced neuroinflammation by regulating Nrf2 signaling pathway.
基金National Natural Science Foundation of China(82030124)National Natural Science Foundation of China(82174015)Science and Technology Innovation Project of China Academy of Traditional Chinese Medicine(CI2021A04609)。
文摘OBJECTIVE To investigate the regulatory effects of icariin(ICA)on cardiac micro⁃vascular endothelial cells(CMEC)after oxygenglucose deprivation reperfusion(OGD/R)injury.METHODS CMEC were subjected to OGD/R treatment to construct a myocardial ischemiareperfusion model,and were divided into normal,model,low(10μmol·L^(-1)),medium(20μmol·L^(-1))and high(40μmol·L^(-1))ICA group,and high ICA+inhibitor group(40μmol·L^(-1)+20 nmol·L^(-1)).CCK-8 assay was used to assess the protective ability of ICA against CMEC,and cell migration assay and tube-formation assay were used to detect the migration and generation ability of CMEC.The TCMSP database,Swiss-Target database and literature mining methods were used to col⁃lect ICA-related targets,the GeneCards data⁃base was used to collect target genes related to myocardial ischemia/reperfusion,and Cytoscape 3.8.0 software was used to construct a"drug-tar⁃get-disease"network.The potential targets were imported into STRING 11.5 database to obtain the PPI network.GO and KEGG enrichment analyses were performed on the potential targets using the DAVID database.Molecular docking was performed using AutoDock-vina 1.1.2 soft⁃ware.Western blot detected the expression of related proteins.RESULTS After CMEC was subjected to OGD/R treatment,ICA had a protec⁃tive effect at 10^(-1)60μmol·L^(-1);the results of the cell migration assay showed that each group of ICA could promote the migratory effect of CMEC(P<0.01,P<0.01);and the results of tube-for⁃mation assay showed that each group of ICA could significantly promote the generation of branches(P<0.01)and the capillary length exten⁃sion(P<0.05).Network pharmacology collected a total of 23 ICA action targets,1500 disease tar⁃gets and 12 key targets.GO function enrichment analysis found 85 results.KEGG pathway enrich⁃ment analysis found 53 results,involving AGERAGE signaling pathway,sphingolipid signaling pathway and VEGF signaling pathway.Molecu⁃lar docking results showed that ICA had better binding with core targets PRKCB,PRKCA and PTGS2.Western blot results showed that ICA could regulate the expression of PRKCB,PRKCA and PTGS2 proteins.The results of cell migra⁃tion assay,tube-formation assay and protein expression were reversed after addition of PKC inhibitor.CONCLUSION The potential mecha⁃nism of action of ICA against myocardial isch⁃emia-reperfusion injury may be related to the reg⁃ulation of processes such as CMEC migration and angiogenesis,and it functions through the key target gene PKC.
