BACKGROUND Gestational diabetes mellitus(GDM)is associated with a heightened level of oxidative stress,which is characterized by the overproduction of reactive oxygen species(ROS)from mitochondria.Previous studies sho...BACKGROUND Gestational diabetes mellitus(GDM)is associated with a heightened level of oxidative stress,which is characterized by the overproduction of reactive oxygen species(ROS)from mitochondria.Previous studies showed that mitochondrial dysfunction is regulated by dynamin-related protein 1(Drp1)and p66Shc in GDM.AIM The aim was to investigate the expression of Drp1 and p66Shc and their possible mechanisms in the pathogenesis of GDM.METHODS A total of 30 pregnant women,15 with GDM and 15 without GDM,were enrolled.Peripheral blood mononuclear cells and placental tissue were collected.The human JEG3 trophoblast cell line was cultivated in 5.5 mmol/L or 30 mmol/L glucose and transfected with wild-type(wt)-p66Shc and p66Shc siRNA.P66Shc and Drp1 mRNA levels were detected by quantitative real-time polymerase chain reaction.The expression of p66Shc and Drp1 was assayed by immunohistochemistry and western blotting.ROS was assayed by dihydroethidium staining.RESULTS The p66Shc mRNA level was increased in the serum(P<0.01)and placentas(P<0.01)of women with GDM,and the expression of Drp1 mRNA and protein were also increased in placentas(P<0.05).In JEG3 cells treated with 30 mmol/L glucose,the mRNA and protein expression of p66Shc and Drp1 were increased at 24 h(both P<0.05),48 h(both P<0.01)and 72 h(both P<0.001).ROS expression was also increased.High levels of Drp1 and ROS expression were detected in JEG3 cells transfected with wt-p66Shc(P<0.01),and low levels were detected in JEG3 cells transfected with p66Shc siRNA(P<0.05).CONCLUSION The upregulated expression of Drp1 and p66shc may contribute to the occurrence and development of GDM.Regulation of the mitochondrial fusion-fission balance could be a novel strategy for GDM treatment.展开更多
BACKGROUND Intervertebral disc degeneration(IDD)is a main contributor to low back pain.Oxidative stress,which is highly associated with the progression of IDD,increases senescence of nucleus pulposus-derived mesenchym...BACKGROUND Intervertebral disc degeneration(IDD)is a main contributor to low back pain.Oxidative stress,which is highly associated with the progression of IDD,increases senescence of nucleus pulposus-derived mesenchymal stem cells(NPMSCs)and weakens the differentiation ability of NPMSCs in degenerated intervertebral discs(IVDs).Quercetin(Que)has been demonstrated to reduce oxidative stress in diverse degenerative diseases.AIM To investigate the role of Que in oxidative stress-induced NPMSC damage and to elucidate the underlying mechanism.METHODS In vitro,NPMSCs were isolated from rat tails.Senescence-associatedβ-galactosidase(SA-β-Gal)staining,cell cycle,reactive oxygen species(ROS),realtime quantitative polymerase chain reaction(RT-qPCR),immunofluorescence,and western blot analyses were used to evaluated the protective effects of Que.Meanwhile the relationship between miR-34a-5p and Sirtuins 1(SIRT1)was evaluated by dual-luciferase reporter assay.To explore whether Que modulates tert-butyl hydroperoxide(TBHP)-induced senescence of NPMSCs via the miR-34a-5p/SIRT1 pathway,we used adenovirus vectors to overexpress and downregulate the expression of miR-34a-5p and used SIRT1 siRNA to knockdown SIRT1 expression.In vivo,a puncture-induced rat IDD model was constructed,and X rays and histological analysis were used to assess whether Que could alleviate IDD in vivo.RESULTS We found that TBHP can cause NPMSCs senescence changes,such as reduced cell proliferation ability,increased SA-β-Gal activity,cell cycle arrest,the accumulation of ROS,and increased expression of senescence-related proteins.While abovementioned senescence indicators were significantly alleviated by Que treatment.Que decreased the expression levels of senescence-related proteins(p16,p21,and p53)and senescence-associated secreted phenotype(SASP),including IL-1β,IL-6,and MMP-13,and it increased the expression of SIRT1.In addition,the protective effects of Que on cell senescence were partially reversed by miR-34a-5p overexpression and SIRT1 knockdown.In vivo,X-ray,and histological analyses indicated that Que alleviated IDD in a punctureinduced rat model.CONCLUSION In summary,the present study provides evidence that Que reduces oxidative stress-induced senescence of NPMSCs via the miR-34a/SIRT1 signaling pathway,suggesting that Que may be a potential agent for the treatment of IDD.展开更多
Objective To investigate the influence of chronic fluorosis on protein kinase Cβ(PKCβ)p66shc signal pathway in the brain of rats,and reveal the molecular mechanism of brain damage.Methods According to body weight by...Objective To investigate the influence of chronic fluorosis on protein kinase Cβ(PKCβ)p66shc signal pathway in the brain of rats,and reveal the molecular mechanism of brain damage.Methods According to body weight by the random number table method thirty SD rats were divided into three groups of 10 each(half females and half males),the normal control group[less than0.5 mg/L of fluorine(prepared with Na F)in展开更多
基金The Scientific Research Fund of Qilu Hospital(Qingdao),No.QDKY2015ZD04.
文摘BACKGROUND Gestational diabetes mellitus(GDM)is associated with a heightened level of oxidative stress,which is characterized by the overproduction of reactive oxygen species(ROS)from mitochondria.Previous studies showed that mitochondrial dysfunction is regulated by dynamin-related protein 1(Drp1)and p66Shc in GDM.AIM The aim was to investigate the expression of Drp1 and p66Shc and their possible mechanisms in the pathogenesis of GDM.METHODS A total of 30 pregnant women,15 with GDM and 15 without GDM,were enrolled.Peripheral blood mononuclear cells and placental tissue were collected.The human JEG3 trophoblast cell line was cultivated in 5.5 mmol/L or 30 mmol/L glucose and transfected with wild-type(wt)-p66Shc and p66Shc siRNA.P66Shc and Drp1 mRNA levels were detected by quantitative real-time polymerase chain reaction.The expression of p66Shc and Drp1 was assayed by immunohistochemistry and western blotting.ROS was assayed by dihydroethidium staining.RESULTS The p66Shc mRNA level was increased in the serum(P<0.01)and placentas(P<0.01)of women with GDM,and the expression of Drp1 mRNA and protein were also increased in placentas(P<0.05).In JEG3 cells treated with 30 mmol/L glucose,the mRNA and protein expression of p66Shc and Drp1 were increased at 24 h(both P<0.05),48 h(both P<0.01)and 72 h(both P<0.001).ROS expression was also increased.High levels of Drp1 and ROS expression were detected in JEG3 cells transfected with wt-p66Shc(P<0.01),and low levels were detected in JEG3 cells transfected with p66Shc siRNA(P<0.05).CONCLUSION The upregulated expression of Drp1 and p66shc may contribute to the occurrence and development of GDM.Regulation of the mitochondrial fusion-fission balance could be a novel strategy for GDM treatment.
基金Supported by the National Natural Science Foundation of China,No.82172462,No.81972136the Traditional Chinese Medicine Science and Technology Development Plan Project of Jiangsu Province,No.YB2020085Cross Cooperation Project of Northern Jiangsu People’s Hospital,No.SBJC21014.
文摘BACKGROUND Intervertebral disc degeneration(IDD)is a main contributor to low back pain.Oxidative stress,which is highly associated with the progression of IDD,increases senescence of nucleus pulposus-derived mesenchymal stem cells(NPMSCs)and weakens the differentiation ability of NPMSCs in degenerated intervertebral discs(IVDs).Quercetin(Que)has been demonstrated to reduce oxidative stress in diverse degenerative diseases.AIM To investigate the role of Que in oxidative stress-induced NPMSC damage and to elucidate the underlying mechanism.METHODS In vitro,NPMSCs were isolated from rat tails.Senescence-associatedβ-galactosidase(SA-β-Gal)staining,cell cycle,reactive oxygen species(ROS),realtime quantitative polymerase chain reaction(RT-qPCR),immunofluorescence,and western blot analyses were used to evaluated the protective effects of Que.Meanwhile the relationship between miR-34a-5p and Sirtuins 1(SIRT1)was evaluated by dual-luciferase reporter assay.To explore whether Que modulates tert-butyl hydroperoxide(TBHP)-induced senescence of NPMSCs via the miR-34a-5p/SIRT1 pathway,we used adenovirus vectors to overexpress and downregulate the expression of miR-34a-5p and used SIRT1 siRNA to knockdown SIRT1 expression.In vivo,a puncture-induced rat IDD model was constructed,and X rays and histological analysis were used to assess whether Que could alleviate IDD in vivo.RESULTS We found that TBHP can cause NPMSCs senescence changes,such as reduced cell proliferation ability,increased SA-β-Gal activity,cell cycle arrest,the accumulation of ROS,and increased expression of senescence-related proteins.While abovementioned senescence indicators were significantly alleviated by Que treatment.Que decreased the expression levels of senescence-related proteins(p16,p21,and p53)and senescence-associated secreted phenotype(SASP),including IL-1β,IL-6,and MMP-13,and it increased the expression of SIRT1.In addition,the protective effects of Que on cell senescence were partially reversed by miR-34a-5p overexpression and SIRT1 knockdown.In vivo,X-ray,and histological analyses indicated that Que alleviated IDD in a punctureinduced rat model.CONCLUSION In summary,the present study provides evidence that Que reduces oxidative stress-induced senescence of NPMSCs via the miR-34a/SIRT1 signaling pathway,suggesting that Que may be a potential agent for the treatment of IDD.
文摘Objective To investigate the influence of chronic fluorosis on protein kinase Cβ(PKCβ)p66shc signal pathway in the brain of rats,and reveal the molecular mechanism of brain damage.Methods According to body weight by the random number table method thirty SD rats were divided into three groups of 10 each(half females and half males),the normal control group[less than0.5 mg/L of fluorine(prepared with Na F)in
