BACKGROUND Treatment of diabetic neuropathy is often limited by side effects.Aucubin,an iridoid glycoside derived from natural plants,exhibits notable anti-inflammatory and antioxidant properties.AIM To investigate th...BACKGROUND Treatment of diabetic neuropathy is often limited by side effects.Aucubin,an iridoid glycoside derived from natural plants,exhibits notable anti-inflammatory and antioxidant properties.AIM To investigate the effects of aucubin on diabetic neuropathic pain(DNP)and glycolysis and inflammation in microglia.METHODS Streptozotocin(STZ)was used to establish a DNP animal model.Blood glucose levels and body weight of mice were measured following STZ administration.Paw withdrawal threshold was calculated for mechanical allodynia.Paw withdrawal latency was recorded for thermal hyperalgesia.The open field test and elevated plus maze was used to assess locomotor activity and anxiety-like behavior.Western blotting was utilized for analysis of protein expression.Immunofluorescence staining was measured for morphometric analysis of microglia.Glycolysis and ATP synthesis in BV-2 cell lines were detected by metabolic extracellular flux analysis.The SwissTargetPrediction and STRING databases were used for comprehensive screening to identify potential target proteins for aucubin.The molecular docking between the possible target proteins and aucubin was investigated using Auto Dock Tool.The BV-2 cell line was transfected with lentiviral AKR1B1-shRNA to further ascertain the function of AKR1B1 in the impact of aucubin on aerobic glycolysis and inflammation during high glucose stimulation.RESULTS Aucubin significantly improved pain and anxiety-like behavior in STZ-induced diabetic mice and restored microglial aerobic glycolysis and inflammation.Several public databases and molecular docking studies suggested that AKR1B1,MMP2 and MMP9 are involved in the effect of aucubin on DNP.Aucubin failed to restore aerobic glycolysis and inflammation in the context of AKR1B1 deficiency.CONCLUSION Aucubin has potential as a therapeutic agent for alleviating DNP by inhibiting expression of AKR1B1.展开更多
Vascular oxidative stress serves as a pathological foundation for various vascular injury-related diseases,including atherosclerosis,hypertension,restenosis,and abdominal aortic aneurysms.Recent studies have indicated...Vascular oxidative stress serves as a pathological foundation for various vascular injury-related diseases,including atherosclerosis,hypertension,restenosis,and abdominal aortic aneurysms.Recent studies have indicated that intestinal flora-derived metabolites,especially phenylacetylglutamine(PAGln)and phenylacetylglycine(PAGly),may contribute to the promotion of thrombosis,heart failure,and other related conditions.Aucubin(AU),an iridoid glycoside,has been shown to exhibit anti-cardiovascular properties.Nevertheless,the precise role and underlying mechanisms by which AU mitigates PAGly-induced vascular injury remain poorly understood.Our results indicated that PAGln/PAGly promoted oxidative stress in vascular endothelial cells(ECs)and vascular smooth muscle cells(VSMCs)in vitro and in vivo.Network pharmacology suggest that AU may possess the capacity to regulate lipid and atherosclerosis,and reactive oxygen species(ROS)processes.We found that AU penetrated the blood vessels and mitigated oxidative stress induced by PAGln/PAGly.Mechanistically,combining the results from intersection analysis between the targets of AU and vascular diseases and molecular docking,we found that tumor necrosis factor(TNF)may be the potential target of AU.Further DARTS and molecular docking analysis demonstrated that AU bound to recombinant TNF-α,and AU could interact with multiple amino acid residues of TNF-α,including Asn-92 and Phe-144.Additionally,PAGly upregulated the level of soluble TNF-α(sTNF-α)in mouse VSMCs and plasma,and promoted the interaction between sTNF-αand TNF receptor 1(TNFR1),whereas AU inhibited this interaction.Both AU and Infliximab,a specific monoclonal antibody of TNF-α,inhibit TNF-α-induced ROS production.In summary,our results revealed that TNF-αis a cellular target of AU,and the interaction between AU and s TNF-αmay mitigate PAGln/PAGly-induced vascular oxidative stress by inhibiting the interaction of TNF-α-TNFR1.展开更多
Rheumatoid arthritis(RA)is a chronic inflammatory autoimmune disease.It is known that aucubin(AU)exerts anti-inflammatory activity,but its effects and mechanisms in RA are unclear.This study investigated the anti-infl...Rheumatoid arthritis(RA)is a chronic inflammatory autoimmune disease.It is known that aucubin(AU)exerts anti-inflammatory activity,but its effects and mechanisms in RA are unclear.This study investigated the anti-inflammatory effects and mechanisms of AU in vivo and in vitro.Human fibroblast-like synoviocyte cells from patients with RA(HFLS-RA),RAW264.7 cells,and MC3T3-E1 cells were used to evaluate the effects of AU on migration,invasion,apoptosis,osteoclast differentiation and production.Immunofluorescence was used to observe nuclear translocation of nuclear factor(NF)-/cB,the double luciferase reporter gene method was used to observe NF-/cB-p65 activity in AU-treated MC3T3-E1 cells.RT-qPCR was used to measure expression of bone metabolism and inflammation-related genes,and western blot was used to measure bone metabolism and NF-a:B protein expression levels.Collagen-induced arthritis(CIA)rat model was used for pharmacodynamics study.Arthritis indexes were measured in the ankle and knee,histological staining and Micro-computed tomography were performed on the ankle joints.Also,inflammatory factor gene expression and the levels of NF-/cB-related proteins were detected as in vitro.AU effectively inhibited HFLS-RA cell migration and invasion,promoted apoptosis,and inhibited RAW264.7 cell differentiation into osteoclasts,as well as inhibited NF-/cB-p65 activity in MC3T3-E1 cells.Notably,AU significantly reduced the gene expression levels of three cell-related inflammatory factors and bone metabolism factors,effectively inhibited the expression of p-lKKafi,p-lA:Ba,and p-p65 proteins.In vivo,AU relieved joint inflammation,reduced related inflammatory factors,and inhibited NF-/cB signaling.It could be used to treat RA-related synovial inflammation and bone destruction through the NF-/cB pathway.展开更多
In this paper,the pharmacological effects of aucubin include anti-inflammatory effect,anti-tumor effect,neuroprotective effect,anti-cardiovascular disease effect,hepatoprotective effect,anti-oxidation effect and anti-...In this paper,the pharmacological effects of aucubin include anti-inflammatory effect,anti-tumor effect,neuroprotective effect,anti-cardiovascular disease effect,hepatoprotective effect,anti-oxidation effect and anti-aging effect,antibacterial effect,anti-diabetes effect and bone protection.The purpose of this study is to provide theoretical basis for the development and clinical application of aucubin.展开更多
Hippocampal neuronal apoptosis accompanied by impairment of cognitive function occurs in primary diabetic encephalopathy. In this study, we investigated the neuroprotective mechanism of the iridoid glycoside, aucubin,...Hippocampal neuronal apoptosis accompanied by impairment of cognitive function occurs in primary diabetic encephalopathy. In this study, we investigated the neuroprotective mechanism of the iridoid glycoside, aucubin, using rats (n=8). Diabetes mellitus was induced in the rats by intraperitoneal (i.p.) injection of streptozotocin (60 mg/kg body weight). After 65 d, half of the DM rats were administered aucubin (5 mg/kg; i.p.) for 15 d, yielding treatment DM+A. A third group of rats received no strepto- zotocin or aucibin, and served as controls (CON). Encephalopathy was assessed using Y-maze be- havioral testing. Rats were euthanized on Day 87, and hippocampi were excised for visual (light and transmission electron microscopic) and immunochemical (Western blot; immunohistochemical) as- sessments of the CA1 subfield for apoptosis and expression of regulatory proteins Bcl-2 and Bax. Treatment responses to all the parameters examined (body weight, plasma glucose, Y-maze error rates, pyramidal cell ultrastructure, proportions of apoptotic cells, levels of expression of Bcl-2 and Bax, and survivability of neuronal cells) were identical: there were highly significant differences between DM and CON groups (P<0.001), but the effects were significantly moderated (P<0.01) in DM+A compared with DM. These findings confirm the association of apoptosis with the encephalopathic effects of diabetes mellitus, and suggest a major role of the expression levels of Bcl-2 and Bax in the regulation of apop- totic cell death. All of the results suggest that aucubin could effectively inhibit apoptosis by modulating the expressions of Bcl-2 and Bax genes.展开更多
To establish a HPLC method to determine the content of aucubin in Eucommia ulmoides seeds,to search for the formation and accumulation of aucubin in Eucommia ulmoides seeds,and to provide foundation for the suitable h...To establish a HPLC method to determine the content of aucubin in Eucommia ulmoides seeds,to search for the formation and accumulation of aucubin in Eucommia ulmoides seeds,and to provide foundation for the suitable harvest season of aucubin in Eucommia ulmoides seeds,the Hypersil GLOD-C18 column(250 mm×4.6 mm,5μm)with temperature at 25 ℃ was used.The mobile phase was methanol-water(8∶92),at a flow rate of 1.0 mL·min-1,and the detective wavelength was 206 nm.The results indicated that the method of HPLC was efficient,and the average recovery of the aucubin in Eucommia ulmoides seeds was 98.61%,RSD=1.14%(n=6).The determination of aucubin was gradually raised with the seed growth,while the formation and accumulation from June 25th to September 5th increased slowly,and the determination of aucubin from September 5th to 25th raised quickily,and then kept relatively stable.Considering the extraction and utilization of aucubin,the date from September 25th to October 25th was the most suitable period for collecting Eucommia ulmoides seeds.展开更多
基金Supported by National Natural Science Foundation of China,No.82001424.
文摘BACKGROUND Treatment of diabetic neuropathy is often limited by side effects.Aucubin,an iridoid glycoside derived from natural plants,exhibits notable anti-inflammatory and antioxidant properties.AIM To investigate the effects of aucubin on diabetic neuropathic pain(DNP)and glycolysis and inflammation in microglia.METHODS Streptozotocin(STZ)was used to establish a DNP animal model.Blood glucose levels and body weight of mice were measured following STZ administration.Paw withdrawal threshold was calculated for mechanical allodynia.Paw withdrawal latency was recorded for thermal hyperalgesia.The open field test and elevated plus maze was used to assess locomotor activity and anxiety-like behavior.Western blotting was utilized for analysis of protein expression.Immunofluorescence staining was measured for morphometric analysis of microglia.Glycolysis and ATP synthesis in BV-2 cell lines were detected by metabolic extracellular flux analysis.The SwissTargetPrediction and STRING databases were used for comprehensive screening to identify potential target proteins for aucubin.The molecular docking between the possible target proteins and aucubin was investigated using Auto Dock Tool.The BV-2 cell line was transfected with lentiviral AKR1B1-shRNA to further ascertain the function of AKR1B1 in the impact of aucubin on aerobic glycolysis and inflammation during high glucose stimulation.RESULTS Aucubin significantly improved pain and anxiety-like behavior in STZ-induced diabetic mice and restored microglial aerobic glycolysis and inflammation.Several public databases and molecular docking studies suggested that AKR1B1,MMP2 and MMP9 are involved in the effect of aucubin on DNP.Aucubin failed to restore aerobic glycolysis and inflammation in the context of AKR1B1 deficiency.CONCLUSION Aucubin has potential as a therapeutic agent for alleviating DNP by inhibiting expression of AKR1B1.
基金supported by the National Key Research and Development Program of China(2022YFF1100300)National Natural Science Foundation of China(32272328)+2 种基金Natural Science Foundation of Hebei Province(H2024206177)Hebei Provincial Key Laboratory of Nutrition and Health(2023YDYY-KF05)College Students’Innovative Entrepreneurial Training Plan Program of Hebei Medical University(USIP2023232)。
文摘Vascular oxidative stress serves as a pathological foundation for various vascular injury-related diseases,including atherosclerosis,hypertension,restenosis,and abdominal aortic aneurysms.Recent studies have indicated that intestinal flora-derived metabolites,especially phenylacetylglutamine(PAGln)and phenylacetylglycine(PAGly),may contribute to the promotion of thrombosis,heart failure,and other related conditions.Aucubin(AU),an iridoid glycoside,has been shown to exhibit anti-cardiovascular properties.Nevertheless,the precise role and underlying mechanisms by which AU mitigates PAGly-induced vascular injury remain poorly understood.Our results indicated that PAGln/PAGly promoted oxidative stress in vascular endothelial cells(ECs)and vascular smooth muscle cells(VSMCs)in vitro and in vivo.Network pharmacology suggest that AU may possess the capacity to regulate lipid and atherosclerosis,and reactive oxygen species(ROS)processes.We found that AU penetrated the blood vessels and mitigated oxidative stress induced by PAGln/PAGly.Mechanistically,combining the results from intersection analysis between the targets of AU and vascular diseases and molecular docking,we found that tumor necrosis factor(TNF)may be the potential target of AU.Further DARTS and molecular docking analysis demonstrated that AU bound to recombinant TNF-α,and AU could interact with multiple amino acid residues of TNF-α,including Asn-92 and Phe-144.Additionally,PAGly upregulated the level of soluble TNF-α(sTNF-α)in mouse VSMCs and plasma,and promoted the interaction between sTNF-αand TNF receptor 1(TNFR1),whereas AU inhibited this interaction.Both AU and Infliximab,a specific monoclonal antibody of TNF-α,inhibit TNF-α-induced ROS production.In summary,our results revealed that TNF-αis a cellular target of AU,and the interaction between AU and s TNF-αmay mitigate PAGln/PAGly-induced vascular oxidative stress by inhibiting the interaction of TNF-α-TNFR1.
基金This work was supported by the Natural Science Foundation of China(No.81773922)Shanghai Natural Science Foundation(No.19ZR1452000).
文摘Rheumatoid arthritis(RA)is a chronic inflammatory autoimmune disease.It is known that aucubin(AU)exerts anti-inflammatory activity,but its effects and mechanisms in RA are unclear.This study investigated the anti-inflammatory effects and mechanisms of AU in vivo and in vitro.Human fibroblast-like synoviocyte cells from patients with RA(HFLS-RA),RAW264.7 cells,and MC3T3-E1 cells were used to evaluate the effects of AU on migration,invasion,apoptosis,osteoclast differentiation and production.Immunofluorescence was used to observe nuclear translocation of nuclear factor(NF)-/cB,the double luciferase reporter gene method was used to observe NF-/cB-p65 activity in AU-treated MC3T3-E1 cells.RT-qPCR was used to measure expression of bone metabolism and inflammation-related genes,and western blot was used to measure bone metabolism and NF-a:B protein expression levels.Collagen-induced arthritis(CIA)rat model was used for pharmacodynamics study.Arthritis indexes were measured in the ankle and knee,histological staining and Micro-computed tomography were performed on the ankle joints.Also,inflammatory factor gene expression and the levels of NF-/cB-related proteins were detected as in vitro.AU effectively inhibited HFLS-RA cell migration and invasion,promoted apoptosis,and inhibited RAW264.7 cell differentiation into osteoclasts,as well as inhibited NF-/cB-p65 activity in MC3T3-E1 cells.Notably,AU significantly reduced the gene expression levels of three cell-related inflammatory factors and bone metabolism factors,effectively inhibited the expression of p-lKKafi,p-lA:Ba,and p-p65 proteins.In vivo,AU relieved joint inflammation,reduced related inflammatory factors,and inhibited NF-/cB signaling.It could be used to treat RA-related synovial inflammation and bone destruction through the NF-/cB pathway.
基金Talent Training Program by the Central Government for the Reform and Development of Local Universities(2020GSP16)Innovation and Entrepreneurship Project for College Students(202110223003).
文摘In this paper,the pharmacological effects of aucubin include anti-inflammatory effect,anti-tumor effect,neuroprotective effect,anti-cardiovascular disease effect,hepatoprotective effect,anti-oxidation effect and anti-aging effect,antibacterial effect,anti-diabetes effect and bone protection.The purpose of this study is to provide theoretical basis for the development and clinical application of aucubin.
基金the National Natural Science Foundation of China (Grant No. 30371053)the National Outstanding Youth Foundation of China (Grant No. 30125034)the Scientific and Technological Plan Project of Dalian City (Grant No. 2003B3NS024)
文摘Hippocampal neuronal apoptosis accompanied by impairment of cognitive function occurs in primary diabetic encephalopathy. In this study, we investigated the neuroprotective mechanism of the iridoid glycoside, aucubin, using rats (n=8). Diabetes mellitus was induced in the rats by intraperitoneal (i.p.) injection of streptozotocin (60 mg/kg body weight). After 65 d, half of the DM rats were administered aucubin (5 mg/kg; i.p.) for 15 d, yielding treatment DM+A. A third group of rats received no strepto- zotocin or aucibin, and served as controls (CON). Encephalopathy was assessed using Y-maze be- havioral testing. Rats were euthanized on Day 87, and hippocampi were excised for visual (light and transmission electron microscopic) and immunochemical (Western blot; immunohistochemical) as- sessments of the CA1 subfield for apoptosis and expression of regulatory proteins Bcl-2 and Bax. Treatment responses to all the parameters examined (body weight, plasma glucose, Y-maze error rates, pyramidal cell ultrastructure, proportions of apoptotic cells, levels of expression of Bcl-2 and Bax, and survivability of neuronal cells) were identical: there were highly significant differences between DM and CON groups (P<0.001), but the effects were significantly moderated (P<0.01) in DM+A compared with DM. These findings confirm the association of apoptosis with the encephalopathic effects of diabetes mellitus, and suggest a major role of the expression levels of Bcl-2 and Bax in the regulation of apop- totic cell death. All of the results suggest that aucubin could effectively inhibit apoptosis by modulating the expressions of Bcl-2 and Bax genes.
文摘To establish a HPLC method to determine the content of aucubin in Eucommia ulmoides seeds,to search for the formation and accumulation of aucubin in Eucommia ulmoides seeds,and to provide foundation for the suitable harvest season of aucubin in Eucommia ulmoides seeds,the Hypersil GLOD-C18 column(250 mm×4.6 mm,5μm)with temperature at 25 ℃ was used.The mobile phase was methanol-water(8∶92),at a flow rate of 1.0 mL·min-1,and the detective wavelength was 206 nm.The results indicated that the method of HPLC was efficient,and the average recovery of the aucubin in Eucommia ulmoides seeds was 98.61%,RSD=1.14%(n=6).The determination of aucubin was gradually raised with the seed growth,while the formation and accumulation from June 25th to September 5th increased slowly,and the determination of aucubin from September 5th to 25th raised quickily,and then kept relatively stable.Considering the extraction and utilization of aucubin,the date from September 25th to October 25th was the most suitable period for collecting Eucommia ulmoides seeds.
