Iron overload injury is considered to be a part of blood stasis syndrome of arthralgia in traditional Chinese medicine.Its primary therapies include clearing heat and detoxification,activating blood circulation,and re...Iron overload injury is considered to be a part of blood stasis syndrome of arthralgia in traditional Chinese medicine.Its primary therapies include clearing heat and detoxification,activating blood circulation,and removing blood stasis.Lonicera japonica flos(LJF)has long been known as an excellent antipyretic and antidote.Luteoloside(Lut)is one of the main components of LJF and exhibits antioxidant,anti-inflammatory,and cytoprotective properties.However,the protection of Lut against iron overload injury and its underlying mechanisms remain unclear.Therefore,HUVECs were exposed to 50μmol·L^(−1)iron dextran for 48 h to establish an iron overload damage model and the effects of Lut were assessed.Our results showed that 20μmol·L^(−1)Lut not only increased cell viability and weakened LDH activity,but also significantly up-regulated DDAHⅡexpression and activity,increased p-eNOS/eNOS ratio and NO content,and reduced ADMA content in HUVECs exposed to iron overload.Furthermore,Lut significantly attenuated intracellular/mitochondrial ROS generation,improved SOD,CAT,and GSH-Px activities,reduced MDA content,maintained MMP,inhibited mPTP opening,prevented cyt c from mitochondria released into cytoplasm,reduced cleaved-caspase3 expression,and ultimately decreased cell apoptosis induced by iron overload.The effects of Lut were similar to those of L-arginine(an ADMA competitive substrate),cyclosporin A(a mPTP blocker agent),and edaravone(a free radical scavenger)as positive controls.However,addition of pAD/DDAHⅡ-shRNA adenovirus reversed the above beneficial effects of Lut.In conclusion,Lut can protect HUVECs against iron overload injury via the ROS/ADMA/DDAHⅡ/eNOS/NO pathway.The mitochondria are the target organelles of Lut’s protective effects.展开更多
Objective:To explore the mechanism of Profilin-1 in regulating eNOS/NO pathway and its role in the development of myocardial hypertrophy.Methods:Spontaneously hypertensive rats(SHR) aged 5 weeks were injected with dif...Objective:To explore the mechanism of Profilin-1 in regulating eNOS/NO pathway and its role in the development of myocardial hypertrophy.Methods:Spontaneously hypertensive rats(SHR) aged 5 weeks were injected with different adenovirus vectors to induce Profilin-1expression knockdown(SHR-I) or over express(SHR-H) or to use as control(SHR-C).All these treatment were compared with Wistar-Kyoto rats(SKY) treated with control adenovirus vectors(WKY-C).The same injection was executed at the sixth week during the experiment of 12 weeks.After experiment,the left ventricular weight-to-heart weight ratio(LVW/HW)and left ventricular long axis(LVLA) were measured.Meanwhile.NO contents in blood and myocardium,Profilin-1,eNOS and Caveolin-3 niRNA and protein levels and phosphorylated eNOS(P-eNOS) protein level in myocardium were determined.Results:Compared with WKY-C group,the SHR-C group was statistically higher in LVW/HW(0.79±0.03).LVLA(11.82±0.58 mm) and Profilin-1 niRNA and prolein level(P<0.05).but lower in NO content[(18.63±6.23) μmol/L| in blood and[(2.71±0.17) μmol/L]in myocardium).eNOS activity and Caveolin-3 expression(P<0.05).The over expressing Profilin-1 led SHR-H group to a higher value of LVW/HW[(0.93±0.03) mm and LVLA(14.17±0.69) mm]in comparison with SHR-C group(P<0.05).and to a lower value of NO content(in myocardium).eNOS activity and Caveolin-3 expression(P<0.05):however,this phenomenon was reversed by the knockdown Profilin-1 expression(SHR-I group).Conclusions:Profilin-1 expression,being negative in regulating Caveolin-3 expression and eNOS/NO pathway activity,promotes the development of myocardial hypertrophy which can be reversed by Profilin-1 silencing.展开更多
Background:American ginseng has been used in the food processing and pharmaceutical industry as a medicinal plant with both nutritional value and economic benefit.Panax quinquefolius saponins(PQS),the main active comp...Background:American ginseng has been used in the food processing and pharmaceutical industry as a medicinal plant with both nutritional value and economic benefit.Panax quinquefolius saponins(PQS),the main active component,have significant antioxidant,neuroprotective,and cardioprotective effects.Clinically,myocardial ischemia(MI)and depression often interact,which has increasing morbidity and mortality rates.However,the mechanism of PQS on MI with depression remains unclear.Methods:The study employed both in vivo and in vitro experiments.Depression-like behaviour changes and cardiac function were observed in mice with MI and depression induced by a high-fat diet(HFD)and intraperitoneal injection of isoproterenol(ISO)plus chronic unpredictable mild stress(CUMS).Both ISO-exposed H9c2 cells and corticosterone(CORT)-induced HT22 cells were selected for in vitro experiments.Biochemical indices and PI3K/Akt/eNOS pathway-related proteins were measured through enzyme-linked immunosorbent assay(ELISA)and Western blotting.Results:PQS significantly improved depression-like behaviour and heart damage in mice and substantially increased H9c2 and HT22 cell activities in vitro.Western blotting analysis showed that PQS could dramatically reverse the changes in the PI3K/AKT/eNOS signalling pathway both in vivo and in vitro.In addition,applying the PI3K inhibitor LY294002 weakened the neuroprotective and cardioprotective effects of PQS.Conclusion:PQS can effectively improve MI with depression,probably through activating PI3K/Akt/eNOS pathway.展开更多
Objective:To establish a mouse model of homocysteine(Hcy)-induced coronary microvascular dysfunction(CMD),and to evaluate the therapeutic efficacy of Shexiang Tongxin dropping pill(STDP)and elucidate its underlying me...Objective:To establish a mouse model of homocysteine(Hcy)-induced coronary microvascular dysfunction(CMD),and to evaluate the therapeutic efficacy of Shexiang Tongxin dropping pill(STDP)and elucidate its underlying mechanisms.Methods:The chemical composition and quality of STDP were characterized using ultra-high performance liquid chromatography,and its absorbed components were identified using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry.CMD was induced in C57BL/6J mice by feeding a 3%methionine diet for four weeks.STDP efficacy was evaluated using laser speckle perfusion imaging,tomato lectin staining,and quantification of plasma nitric oxide(NO),reactive oxygen species(ROS),and endothelial adhesion molecules(intercellular cell adhesion molecule-1[ICAM-1],vascular cell adhesion molecule-1[VCAM-1]).Network pharmacology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed to identify potential targets and regulatory pathways.An in vitro Hcy-induced endothelial injury model was used to validate the effects of STDP on cell viability,NO production,and activation of phosphatidylinositol 3-kinase/protein kinase B/endothelial nitric oxide synthase(PI3K/Akt/eNOS)pathway.Results:STDP was stable,with 180 constituents identified in the preparation and 30 absorbed components in plasma.STDP treatment restored perfusion,increased plasma NO,decreased ROS,and downregulated ICAM-1 and VCAM-1.Network analysis identified 152 putative targets,highlighting the PI3K/Akt pathway as the central,with PIK3CA,AKT1,and NOS3 as key nodes.In vitro,STDP enhanced cell viability,NO production,and PI3K/Akt/eNOS phosphorylation,these effects were abolished by pharmacological inhibition of PI3K and eNOS.Conclusion:A 3%methionine diet for four weeks effectively induces CMD in C57BL/6J mice.STDP,rich in bioactive components,alleviates Hcy-induced CMD by activating the PI3K/Akt/eNOS pathway,thereby improving endothelial function and microvascular perfusion.These findings support STDP as a promising therapeutic candidate for CMD management.展开更多
LVLPGE is an angiotensin-converting enzyme(ACE)inhibitory peptide extracted from broccoli protein hydrolysates.The antihypertensive mechanism of LVLPGE was explored in the present study.The results indicated that the ...LVLPGE is an angiotensin-converting enzyme(ACE)inhibitory peptide extracted from broccoli protein hydrolysates.The antihypertensive mechanism of LVLPGE was explored in the present study.The results indicated that the IC50 value of LVLPGE was 19.49μM,and its binding to ACE occurred via a static fluorescence quenching involving the secondary structure changes of ACE.In addition,LVLPGE competitively bound to the active site of ACE through hydrogen bonding.Furthermore,LVLPGE markedly enhanced the levels of the vasodilator nitric oxide(NO)in the Ang I-induced HUVECs model,suppressed the expression of vasoconstrictor endothelin-1(ET-1),and increased the phosphorylation of eNOS.Moreover,LVLPGE was found to enhance the production of cyclic guanosine monophosphate(cGMP),which leading to a subsequent decrease in intracellular calcium concentration,ultimately facilitating the relaxation of vascular smooth muscles and lowering the blood pressure.These results demonstrated that LVLPGE achieves the antihypertensive effects by inhibiting ACE activity and activating the eNOS/NO/cGMP signaling pathway.This study provides new insights into the antihypertensive mechanism of LVLPGE and contributes to the high-value utilization of broccoli resources.展开更多
基金the National Natural Science Foundation of China(Nos.21467017,81660583,81673431,and 81803534)Jiangxi Applied Research and Cultivation Program(No.20181BBG78059).
文摘Iron overload injury is considered to be a part of blood stasis syndrome of arthralgia in traditional Chinese medicine.Its primary therapies include clearing heat and detoxification,activating blood circulation,and removing blood stasis.Lonicera japonica flos(LJF)has long been known as an excellent antipyretic and antidote.Luteoloside(Lut)is one of the main components of LJF and exhibits antioxidant,anti-inflammatory,and cytoprotective properties.However,the protection of Lut against iron overload injury and its underlying mechanisms remain unclear.Therefore,HUVECs were exposed to 50μmol·L^(−1)iron dextran for 48 h to establish an iron overload damage model and the effects of Lut were assessed.Our results showed that 20μmol·L^(−1)Lut not only increased cell viability and weakened LDH activity,but also significantly up-regulated DDAHⅡexpression and activity,increased p-eNOS/eNOS ratio and NO content,and reduced ADMA content in HUVECs exposed to iron overload.Furthermore,Lut significantly attenuated intracellular/mitochondrial ROS generation,improved SOD,CAT,and GSH-Px activities,reduced MDA content,maintained MMP,inhibited mPTP opening,prevented cyt c from mitochondria released into cytoplasm,reduced cleaved-caspase3 expression,and ultimately decreased cell apoptosis induced by iron overload.The effects of Lut were similar to those of L-arginine(an ADMA competitive substrate),cyclosporin A(a mPTP blocker agent),and edaravone(a free radical scavenger)as positive controls.However,addition of pAD/DDAHⅡ-shRNA adenovirus reversed the above beneficial effects of Lut.In conclusion,Lut can protect HUVECs against iron overload injury via the ROS/ADMA/DDAHⅡ/eNOS/NO pathway.The mitochondria are the target organelles of Lut’s protective effects.
基金funded by Key Disciplines Group Construction Project of Pudong Health Bureau of Shanghai(No.PWZxq2014-01)Natural Science Foundation of China(No.81401428)the fund of the Academic Leaders Training Program of Pudong Health Bureau of Shanghai(No.PWRd2013-02)
文摘Objective:To explore the mechanism of Profilin-1 in regulating eNOS/NO pathway and its role in the development of myocardial hypertrophy.Methods:Spontaneously hypertensive rats(SHR) aged 5 weeks were injected with different adenovirus vectors to induce Profilin-1expression knockdown(SHR-I) or over express(SHR-H) or to use as control(SHR-C).All these treatment were compared with Wistar-Kyoto rats(SKY) treated with control adenovirus vectors(WKY-C).The same injection was executed at the sixth week during the experiment of 12 weeks.After experiment,the left ventricular weight-to-heart weight ratio(LVW/HW)and left ventricular long axis(LVLA) were measured.Meanwhile.NO contents in blood and myocardium,Profilin-1,eNOS and Caveolin-3 niRNA and protein levels and phosphorylated eNOS(P-eNOS) protein level in myocardium were determined.Results:Compared with WKY-C group,the SHR-C group was statistically higher in LVW/HW(0.79±0.03).LVLA(11.82±0.58 mm) and Profilin-1 niRNA and prolein level(P<0.05).but lower in NO content[(18.63±6.23) μmol/L| in blood and[(2.71±0.17) μmol/L]in myocardium).eNOS activity and Caveolin-3 expression(P<0.05).The over expressing Profilin-1 led SHR-H group to a higher value of LVW/HW[(0.93±0.03) mm and LVLA(14.17±0.69) mm]in comparison with SHR-C group(P<0.05).and to a lower value of NO content(in myocardium).eNOS activity and Caveolin-3 expression(P<0.05):however,this phenomenon was reversed by the knockdown Profilin-1 expression(SHR-I group).Conclusions:Profilin-1 expression,being negative in regulating Caveolin-3 expression and eNOS/NO pathway activity,promotes the development of myocardial hypertrophy which can be reversed by Profilin-1 silencing.
基金supported by the study on the material basis and mechanism of action of American ginseng in the treatment of myocardial ischaemia comorbid depression(2024JH2/102500059).
文摘Background:American ginseng has been used in the food processing and pharmaceutical industry as a medicinal plant with both nutritional value and economic benefit.Panax quinquefolius saponins(PQS),the main active component,have significant antioxidant,neuroprotective,and cardioprotective effects.Clinically,myocardial ischemia(MI)and depression often interact,which has increasing morbidity and mortality rates.However,the mechanism of PQS on MI with depression remains unclear.Methods:The study employed both in vivo and in vitro experiments.Depression-like behaviour changes and cardiac function were observed in mice with MI and depression induced by a high-fat diet(HFD)and intraperitoneal injection of isoproterenol(ISO)plus chronic unpredictable mild stress(CUMS).Both ISO-exposed H9c2 cells and corticosterone(CORT)-induced HT22 cells were selected for in vitro experiments.Biochemical indices and PI3K/Akt/eNOS pathway-related proteins were measured through enzyme-linked immunosorbent assay(ELISA)and Western blotting.Results:PQS significantly improved depression-like behaviour and heart damage in mice and substantially increased H9c2 and HT22 cell activities in vitro.Western blotting analysis showed that PQS could dramatically reverse the changes in the PI3K/AKT/eNOS signalling pathway both in vivo and in vitro.In addition,applying the PI3K inhibitor LY294002 weakened the neuroprotective and cardioprotective effects of PQS.Conclusion:PQS can effectively improve MI with depression,probably through activating PI3K/Akt/eNOS pathway.
基金supported by the National Key Research and Development Program of China(2022YFC3500100)the National Natural Science Foundation of China(82230126 and U24A20800)+2 种基金the National Science and Technology Major Project of the Ministry of Science and Technology of China(2023ZD0502600)National Science Fund for Excellent Young Scholars(82222075)the Incubation Program for the Science and Technology Development of Chinese Medicine Guangdong Laboratory(Project HQL2024PZ045 and HQCML).
文摘Objective:To establish a mouse model of homocysteine(Hcy)-induced coronary microvascular dysfunction(CMD),and to evaluate the therapeutic efficacy of Shexiang Tongxin dropping pill(STDP)and elucidate its underlying mechanisms.Methods:The chemical composition and quality of STDP were characterized using ultra-high performance liquid chromatography,and its absorbed components were identified using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry.CMD was induced in C57BL/6J mice by feeding a 3%methionine diet for four weeks.STDP efficacy was evaluated using laser speckle perfusion imaging,tomato lectin staining,and quantification of plasma nitric oxide(NO),reactive oxygen species(ROS),and endothelial adhesion molecules(intercellular cell adhesion molecule-1[ICAM-1],vascular cell adhesion molecule-1[VCAM-1]).Network pharmacology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed to identify potential targets and regulatory pathways.An in vitro Hcy-induced endothelial injury model was used to validate the effects of STDP on cell viability,NO production,and activation of phosphatidylinositol 3-kinase/protein kinase B/endothelial nitric oxide synthase(PI3K/Akt/eNOS)pathway.Results:STDP was stable,with 180 constituents identified in the preparation and 30 absorbed components in plasma.STDP treatment restored perfusion,increased plasma NO,decreased ROS,and downregulated ICAM-1 and VCAM-1.Network analysis identified 152 putative targets,highlighting the PI3K/Akt pathway as the central,with PIK3CA,AKT1,and NOS3 as key nodes.In vitro,STDP enhanced cell viability,NO production,and PI3K/Akt/eNOS phosphorylation,these effects were abolished by pharmacological inhibition of PI3K and eNOS.Conclusion:A 3%methionine diet for four weeks effectively induces CMD in C57BL/6J mice.STDP,rich in bioactive components,alleviates Hcy-induced CMD by activating the PI3K/Akt/eNOS pathway,thereby improving endothelial function and microvascular perfusion.These findings support STDP as a promising therapeutic candidate for CMD management.
基金Grants from National Key R&D Program of China(2022YFF11002)Zhejiang Provincial Natural Science Foundation(LQ23C200009)National Natural Science Foundation of China(32302052).
文摘LVLPGE is an angiotensin-converting enzyme(ACE)inhibitory peptide extracted from broccoli protein hydrolysates.The antihypertensive mechanism of LVLPGE was explored in the present study.The results indicated that the IC50 value of LVLPGE was 19.49μM,and its binding to ACE occurred via a static fluorescence quenching involving the secondary structure changes of ACE.In addition,LVLPGE competitively bound to the active site of ACE through hydrogen bonding.Furthermore,LVLPGE markedly enhanced the levels of the vasodilator nitric oxide(NO)in the Ang I-induced HUVECs model,suppressed the expression of vasoconstrictor endothelin-1(ET-1),and increased the phosphorylation of eNOS.Moreover,LVLPGE was found to enhance the production of cyclic guanosine monophosphate(cGMP),which leading to a subsequent decrease in intracellular calcium concentration,ultimately facilitating the relaxation of vascular smooth muscles and lowering the blood pressure.These results demonstrated that LVLPGE achieves the antihypertensive effects by inhibiting ACE activity and activating the eNOS/NO/cGMP signaling pathway.This study provides new insights into the antihypertensive mechanism of LVLPGE and contributes to the high-value utilization of broccoli resources.
