Objective:To investigate the mechanism of fire needling in improving autophagy and oxidative stress in rats with cervical spondylosis of vertebral artery type(CSA)by regulating protein kinase B(PKB/Akt)/mammalian targ...Objective:To investigate the mechanism of fire needling in improving autophagy and oxidative stress in rats with cervical spondylosis of vertebral artery type(CSA)by regulating protein kinase B(PKB/Akt)/mammalian target of rapamycin(mTOR)signaling pathway.Methods:The rats were randomly divided into a sham-operation group(Sham group)and a model group.After successful modeling,the rats were randomly divided into a CSA group,a fire needling group,and a fire needling+insulin-like growth factor 1(IGF-1)group.No intervention was performed in the Sham and CSA groups;the fire needling group received fire needling intervention;the fire needling+IGF-1 group received both fire needling and intraperitoneal injection of IGF-1 solution intervention.The inclined plate test was used to detect the exercise ability of rats.Laser Doppler was used to detect the blood flow in the pia mater microcirculation.Multi-mode high-frequency acoustic was used to detect the blood flow velocity of both sides of the vertebral artery.The serum levels of endothelin-1(ET-1),nitric oxide(NO),superoxide dismutase(SOD),and malondialdehyde(MDA)were measured.A transmission electron microscope was used to observe vertebral artery autophagosomes.Western blotting was used to detect the ratios of phosphorylated(phospho)-phosphoinositide 3-kinase(PI3K)/PI3K,phospho-Akt/Akt,phospho-mTOR/mTOR,autophagy-related proteins(Beclin-1 and p62),and the ratios of microtubule-associated protein 1A/1B light chain 3(LC3Ⅱ/LC3Ⅰ)in vertebral artery tissues.Results:Compared to the Sham group,the inclination angle of the inclined plate,pia mater microcirculation blood flow,blood flow velocity of the left vertebral artery(LVA),right vertebral artery(RVA),NO level,and SOD activity were significantly decreased(P<0.05),and the serum ET-1 and MDA levels were significantly increased(P<0.05)in the CSA group.Compared to the CSA group,the inclination angle of the inclined plate,blood flow of pia mater microcirculation,blood flow velocity of the LVA and RVA,NO level,and SOD activity were significantly increased(P<0.05),and the serum ET-1 and MDA levels were significantly decreased(P<0.05)in the fire needling group.The inclination angle of the inclined plate,blood flow of pia mater microcirculation,blood flow velocity of the LVA and RVA,NO level,and SOD activity in the fire needling+IGF-1 group were significantly lower than those in the fire needling group(P<0.05),and the serum ET-1 and MDA levels were higher than those in the fire needling group(P<0.05).Compared to the Sham group,a large number of autophagosomes and autophagy degradation vesicles were found in the vertebral artery tissues of the CSA group.Compared to the CSA group,autophagosomes and autophagy degradation vesicles in rat vertebral artery tissues of the fire needling group were significantly reduced.Compared to the fire needling group,the autophagosomes and autophagy degradation vesicles in the vertebral artery tissues of the fire needling+IGF-1 group were increased significantly.The expression ratios of phospho-PI3K/PI3K,phospho-Akt/Akt,phospho-mTOR/mTOR,LC3Ⅱ/LC3Ⅰ,and Beclin protein expression in rat vertebral artery tissues of the CSA group were higher than those in the Sham group(P<0.05),and p62 protein expression was lower than that in the Sham group(P<0.05).The above expression ratios in rat vertebral artery tissues of the fire needling group were lower than those of the CSA group(P<0.05),and p62 protein expression was higher than that of the CSA group(P<0.05).The above protein expression ratios in rat vertebral artery tissues of the fire needling+IGF-1 group were higher than those of the fire needling group(P<0.05),and p62 protein expression was lower than that of the fire needling group(P<0.05).Conclusion:Fire needling can reduce oxidative stress levels by promoting autophagy in CSA rats.The mechanism may be related to the inhibition of PI3K/Akt/mTOR signaling pathway activation.展开更多
Nekemias megalophylla is a popular folk tea consumed by people in the Western Hubei(China)of which ampelopsin(AMP)is the main active ingredient.In this study,we investigated the effect of AMP on cervical cancer and ex...Nekemias megalophylla is a popular folk tea consumed by people in the Western Hubei(China)of which ampelopsin(AMP)is the main active ingredient.In this study,we investigated the effect of AMP on cervical cancer and explored its mechanism of action,focusing on apoptosis and autophagy.Firstly,we verified that AMP strongly inhibited the growth of C-33A cells and observed apoptosis and autophagy phenomenon in vivo,and found that AMP induces C-33A cell apoptosis via death receptor or mitochondrial pathways.The results also indicated that AMP-induced autophagy occurs via the PI3K/Akt/m TOR pathway.Secondly,when autophagy was inhibited,the AMP-induced apoptosis of C-33A cells was strengthened,when apoptosis was inhibited,the AMP-induced autophagy of C-33A cells was strengthened.PI3K/Akt/m TOR pathway activation enhances AMP-induced apoptosis in C-33A cells,while its inhibition strengthens AMP-induced autophagy.Finally,we confirmed that AMP inhibited cell growth and induced apoptosis and autophagy of C-33A cells in an in vivo nude mouse model of C-33A tumor xenografts.These results elucidate that AMP bidirectionally regulates apoptosis and autophagy in human cervical cancer C-33A cells by mediating the PI3K/Akt/m TOR pathway.展开更多
Objective:To investigate the anti-atherosclerosis effect of chikusetsusaponinⅣ(CSⅣ)against high-fat diet-induced atherosclerosis in rats.Methods:A high-fat diet was used for the induction of atherosclerosis in rats,...Objective:To investigate the anti-atherosclerosis effect of chikusetsusaponinⅣ(CSⅣ)against high-fat diet-induced atherosclerosis in rats.Methods:A high-fat diet was used for the induction of atherosclerosis in rats,and the rats received oral CSⅣor atorvastatin.The body weight,organ weights,food intake,calorie intake,lipid parameters,3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA)/mevalonate ratio,collagen,free fatty acid,cardiac parameters,apolipoprotein(A and B),antioxidant parameters,inflammatory cytokines,and inflammatory parameters were assessed.The mRNA expressions of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6,IL-17,PI3K,AKT,and mTOR were estimated.Results:CSⅣsignificantly modulated food intake,body weight,organ weight(liver,kidney,and heart),and calories(P<0.05).Total cholesterol,triglycerides,very low-density lipoprotein cholesterol,low-density lipoprotein cholesterol,cardiovascular risk index-1,and cardiovascular risk index-2 were decreased,while high-density lipoprotein cholesterol and anti-atherogenic index were increased significantly in the CSⅣgroup(P<0.05).Besides,CSⅣsignificantly restored the level of HMG-CoA/mevalonate ratio,collagen,free fatty acid,cardiac parameters(creatinine kinase-MB,lactate dehydrogenase,cTnT,cTnI),apolipoprotein(apolipoprotein A and apolipoprotein B),antioxidant parameters(MDA,CAT,GPx,GSH,SOD),inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10),inflammatory parameters(COX-2,TGF-β,NF-κB),intercellular adhesion molecule-1,vascular cell adhesion molecule-1,and monocyte chemoattractant protein-1.CSⅣalso decreased the mRNA expression of IL-1β,TNF-α,IL-6,IL-17,PI3K,AKT,and mTOR.Conclusions:This study showed the anti-atherosclerosis effect of CSⅣagainst high-fat diet-induced atherosclerosis in rats via alteration of NF-κB/COX-2 and PI3K/AKT/mTOR signaling pathway.展开更多
基金supported by Chongqing Science and Technology Project of Traditional Chinese Medicine Jointly Launched by Science and Health Commission in 2018(重庆市2018年科卫联合中医药科技项目,No.ZY201802027).
文摘Objective:To investigate the mechanism of fire needling in improving autophagy and oxidative stress in rats with cervical spondylosis of vertebral artery type(CSA)by regulating protein kinase B(PKB/Akt)/mammalian target of rapamycin(mTOR)signaling pathway.Methods:The rats were randomly divided into a sham-operation group(Sham group)and a model group.After successful modeling,the rats were randomly divided into a CSA group,a fire needling group,and a fire needling+insulin-like growth factor 1(IGF-1)group.No intervention was performed in the Sham and CSA groups;the fire needling group received fire needling intervention;the fire needling+IGF-1 group received both fire needling and intraperitoneal injection of IGF-1 solution intervention.The inclined plate test was used to detect the exercise ability of rats.Laser Doppler was used to detect the blood flow in the pia mater microcirculation.Multi-mode high-frequency acoustic was used to detect the blood flow velocity of both sides of the vertebral artery.The serum levels of endothelin-1(ET-1),nitric oxide(NO),superoxide dismutase(SOD),and malondialdehyde(MDA)were measured.A transmission electron microscope was used to observe vertebral artery autophagosomes.Western blotting was used to detect the ratios of phosphorylated(phospho)-phosphoinositide 3-kinase(PI3K)/PI3K,phospho-Akt/Akt,phospho-mTOR/mTOR,autophagy-related proteins(Beclin-1 and p62),and the ratios of microtubule-associated protein 1A/1B light chain 3(LC3Ⅱ/LC3Ⅰ)in vertebral artery tissues.Results:Compared to the Sham group,the inclination angle of the inclined plate,pia mater microcirculation blood flow,blood flow velocity of the left vertebral artery(LVA),right vertebral artery(RVA),NO level,and SOD activity were significantly decreased(P<0.05),and the serum ET-1 and MDA levels were significantly increased(P<0.05)in the CSA group.Compared to the CSA group,the inclination angle of the inclined plate,blood flow of pia mater microcirculation,blood flow velocity of the LVA and RVA,NO level,and SOD activity were significantly increased(P<0.05),and the serum ET-1 and MDA levels were significantly decreased(P<0.05)in the fire needling group.The inclination angle of the inclined plate,blood flow of pia mater microcirculation,blood flow velocity of the LVA and RVA,NO level,and SOD activity in the fire needling+IGF-1 group were significantly lower than those in the fire needling group(P<0.05),and the serum ET-1 and MDA levels were higher than those in the fire needling group(P<0.05).Compared to the Sham group,a large number of autophagosomes and autophagy degradation vesicles were found in the vertebral artery tissues of the CSA group.Compared to the CSA group,autophagosomes and autophagy degradation vesicles in rat vertebral artery tissues of the fire needling group were significantly reduced.Compared to the fire needling group,the autophagosomes and autophagy degradation vesicles in the vertebral artery tissues of the fire needling+IGF-1 group were increased significantly.The expression ratios of phospho-PI3K/PI3K,phospho-Akt/Akt,phospho-mTOR/mTOR,LC3Ⅱ/LC3Ⅰ,and Beclin protein expression in rat vertebral artery tissues of the CSA group were higher than those in the Sham group(P<0.05),and p62 protein expression was lower than that in the Sham group(P<0.05).The above expression ratios in rat vertebral artery tissues of the fire needling group were lower than those of the CSA group(P<0.05),and p62 protein expression was higher than that of the CSA group(P<0.05).The above protein expression ratios in rat vertebral artery tissues of the fire needling+IGF-1 group were higher than those of the fire needling group(P<0.05),and p62 protein expression was lower than that of the fire needling group(P<0.05).Conclusion:Fire needling can reduce oxidative stress levels by promoting autophagy in CSA rats.The mechanism may be related to the inhibition of PI3K/Akt/mTOR signaling pathway activation.
基金supported by the Major Science and Technology Project of Hubei Province(2020ACA007)the Scientific and Technological Bureau of Wuhan(2018060401011308).
文摘Nekemias megalophylla is a popular folk tea consumed by people in the Western Hubei(China)of which ampelopsin(AMP)is the main active ingredient.In this study,we investigated the effect of AMP on cervical cancer and explored its mechanism of action,focusing on apoptosis and autophagy.Firstly,we verified that AMP strongly inhibited the growth of C-33A cells and observed apoptosis and autophagy phenomenon in vivo,and found that AMP induces C-33A cell apoptosis via death receptor or mitochondrial pathways.The results also indicated that AMP-induced autophagy occurs via the PI3K/Akt/m TOR pathway.Secondly,when autophagy was inhibited,the AMP-induced apoptosis of C-33A cells was strengthened,when apoptosis was inhibited,the AMP-induced autophagy of C-33A cells was strengthened.PI3K/Akt/m TOR pathway activation enhances AMP-induced apoptosis in C-33A cells,while its inhibition strengthens AMP-induced autophagy.Finally,we confirmed that AMP inhibited cell growth and induced apoptosis and autophagy of C-33A cells in an in vivo nude mouse model of C-33A tumor xenografts.These results elucidate that AMP bidirectionally regulates apoptosis and autophagy in human cervical cancer C-33A cells by mediating the PI3K/Akt/m TOR pathway.
基金funded by the Yancheng Municipal Health Commission 2024 Medical Research Project(YK2024166).
文摘Objective:To investigate the anti-atherosclerosis effect of chikusetsusaponinⅣ(CSⅣ)against high-fat diet-induced atherosclerosis in rats.Methods:A high-fat diet was used for the induction of atherosclerosis in rats,and the rats received oral CSⅣor atorvastatin.The body weight,organ weights,food intake,calorie intake,lipid parameters,3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA)/mevalonate ratio,collagen,free fatty acid,cardiac parameters,apolipoprotein(A and B),antioxidant parameters,inflammatory cytokines,and inflammatory parameters were assessed.The mRNA expressions of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),IL-6,IL-17,PI3K,AKT,and mTOR were estimated.Results:CSⅣsignificantly modulated food intake,body weight,organ weight(liver,kidney,and heart),and calories(P<0.05).Total cholesterol,triglycerides,very low-density lipoprotein cholesterol,low-density lipoprotein cholesterol,cardiovascular risk index-1,and cardiovascular risk index-2 were decreased,while high-density lipoprotein cholesterol and anti-atherogenic index were increased significantly in the CSⅣgroup(P<0.05).Besides,CSⅣsignificantly restored the level of HMG-CoA/mevalonate ratio,collagen,free fatty acid,cardiac parameters(creatinine kinase-MB,lactate dehydrogenase,cTnT,cTnI),apolipoprotein(apolipoprotein A and apolipoprotein B),antioxidant parameters(MDA,CAT,GPx,GSH,SOD),inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10),inflammatory parameters(COX-2,TGF-β,NF-κB),intercellular adhesion molecule-1,vascular cell adhesion molecule-1,and monocyte chemoattractant protein-1.CSⅣalso decreased the mRNA expression of IL-1β,TNF-α,IL-6,IL-17,PI3K,AKT,and mTOR.Conclusions:This study showed the anti-atherosclerosis effect of CSⅣagainst high-fat diet-induced atherosclerosis in rats via alteration of NF-κB/COX-2 and PI3K/AKT/mTOR signaling pathway.
