OBJECTIVE:To investigate the effect of Ruyanneixiao cream(RYNX) on the expression of hypoxia inducible factor-1α(HIF-1α), hexokinase 2(HK2),phosphofructokinase(PFK), and pyruvate kinase M2(PKM2) mRNA and protein in ...OBJECTIVE:To investigate the effect of Ruyanneixiao cream(RYNX) on the expression of hypoxia inducible factor-1α(HIF-1α), hexokinase 2(HK2),phosphofructokinase(PFK), and pyruvate kinase M2(PKM2) mRNA and protein in MCF-10 AT cells and in an animal model of precancerous mammary lesions.METHODS:Following treatment of MCF-10 AT cells with RYNX, tamoxifen(TAM) and YC-1 for 48 h,HIF-1α, HK2, PFK, PKM2 mRNA and protein expression was analyzed.Fifty female SD rats were randomly divided into control, model, TAM, and highand low-dose RYNX groups, with 10 rats in each group.A precancerous mammary lesion model was established for all groups except the control group.High-and low-dose RYNX cream containing TAM was coated on the breasts of animals in the corresponding groups.The rat mammary tissue was removed in the 10 th week and HIF-1α, HK2, PFK,PKM2 mRNA and protein was analyzed.RESULTS:In vitro analyses demonstrated that, compared with the matrix group, HIF-1α, HK2, PFK,PKM2 mRNA and protein expression was significantly decreased in the RYNX group(P < 0.05).Compared with the YC-1 + RYNX group, HK2, PFK,and PKM2 protein expression was significantly reduced in the RYNX group.HIF-1α, HK2, PFK, and PKM2 protein expression was increased significantly in the model group(P < 0.05) compared with the control group, while HIF-1α, HK2, PFK, and PKM2 mRNA and protein expression was significantly decreased in both the high-and low-dose RYNX groups(P < 0.05), with the effect being greater in the high-dose group.CONCLUSION:RYNX can block precancerous breast lesions by decreasing the expression of HK2,PFK, and PKM2 mRNA and protein via inhibition of HIF-1α mRNA and protein overexpression in a dose-dependent manner.展开更多
The Pomacea canaliculata,also known as apple snail,is an invasive species in China,posing a serious threat to agricultural production and public health.To prevent its rampant infestation,we developed a highly efficien...The Pomacea canaliculata,also known as apple snail,is an invasive species in China,posing a serious threat to agricultural production and public health.To prevent its rampant infestation,we developed a highly efficient snail-killing lead compound,1-(4-chlorophenyl)-3-(pyridin-3-yl)urea(PBQ).However,the killing mechanism of PBQ remains unclear.To elucidate this,two novel compounds,B-PBQ and J-PBQ,were developed through structural modification of PBQ and were used,through both protein pull-down assay and confocal laser scanning microscopy imaging,to identify and confirm that phosphofructokinase(PFK)in the apple snail is the primary target protein of PBQ.Further studies revealed that PBQ successfully inhibits the activity,content,and expression of PFK in apple snail,disrupting the glycolysis pathway and leading to abnormal energy production.These disruptions impair the snail's tolerance to low oxygen,resulting in its efficient eradication.These findings offer a promising direction for developing next-generation PFK-targeted snail-killing pesticides.Furthermore,the strategy of combining pull-down assays with optical imaging used in this work provides a novel approach for the exploration of mechanisms of other pesticide candidates.展开更多
Zinc-dependent histone deacetylases(HDACs)are pivotal enzymes governing the epigenetic modulation of gene expression through chromatin remodeling.The dysregulated expression of HDACs is intricately linked to various p...Zinc-dependent histone deacetylases(HDACs)are pivotal enzymes governing the epigenetic modulation of gene expression through chromatin remodeling.The dysregulated expression of HDACs is intricately linked to various pathological conditions,including cancer and inflammation.Histone deacetylase inhibitors(HDACi)have shown therapeutic potential in certain hematologic malignancies.However,the clinical performance of HDACi in solid tumors remains unsatisfactory,and the precise mechanisms of its therapeutic effect in solid tumors has not been fully elucidated.In this study,we identified nucleus-localized PFKL(Liver-type phosphofructokinase),as a key regulator of HDACi efficacy and intracellular epigenetic dynamics.Nuclear PFKL directly binds to classⅠHDACs through interacting with zinc-binding sites,thereby inhibiting HDAC enzymatic activity and promoting intracellular histone acetylation.In addition,the Thr562 residue within PFKL enhances the chelation effect between the zinc-binding group(ZBG)of the HDACi romidepsin and the zinc within the HDACs,further promoting drug efficacy.Based on the mechanism of PFKL facilitates the efficacy of romidepsin,we developed a therapeutic peptide,PFKL-552-572-R8,which significantly enhances the antitumor effect of romidepsin both in vitro and in vivo.Our findings reveal that spatiotemporal regulation confers a moonlight function to PFKL as an endogenous HDAC inhibitor to maintain the stability of epigenetic modifications and highlight PFKL as a promising therapeutic target for enhancing the clinical utility of HDACi in solid tumors.展开更多
Sugars are involved in plant growth,fruit quality,and signaling perception.Therefore,understanding the mechanisms involved in soluble sugar accumulation is essential to understand fruit development.Here,we report that...Sugars are involved in plant growth,fruit quality,and signaling perception.Therefore,understanding the mechanisms involved in soluble sugar accumulation is essential to understand fruit development.Here,we report that Md PFPβ,a pyrophosphatedependent phosphofructokinase gene,regulates soluble sugar accumulation by enhancing the photosynthetic performance and sugar-metabolizing enzyme activities in apple(Malus domestica Borkh.).Biochemical analysis revealed that a basic helix-loop-helix(b HLH)transcription factor,Mdb HLH3,binds to the Md PFPβpromoter and activates its expression,thus promoting soluble sugar accumulation in apple fruit.In addition,Md PFPβoverexpression in tomato influenced photosynthesis and carbon metabolism in the plant.Furthermore,we determined that Mdb HLH3 increases photosynthetic rates and soluble sugar accumulation in apple by activating Md PFPβexpression.Our results thus shed light on the mechanism of soluble sugar accumulation in apple leaves and fruit:Mdb HLH3 regulates soluble sugar accumulation by activating Md PFPβgene expression and coordinating carbohydrate allocation.展开更多
Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to impr...Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to improve migration and survival of bone marrow–derived mesenchymal stem cells and reduce pyroptosis after cardiac arrest,but the specific mechanisms by which hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect against brain injury after cardiac arrest are unknown.To this end,we established an in vitro co-culture model of bone marrow–derived mesenchymal stem cells and oxygen–glucose deprived primary neurons and found that hypoxic preconditioning enhanced the protective effect of bone marrow stromal stem cells against neuronal pyroptosis,possibly through inhibition of the MAPK and nuclear factor κB pathways.Subsequently,we transplanted hypoxia-preconditioned bone marrow–derived mesenchymal stem cells into the lateral ventricle after the return of spontaneous circulation in an 8-minute cardiac arrest rat model induced by asphyxia.The results showed that hypoxia-preconditioned bone marrow–derived mesenchymal stem cells significantly reduced cardiac arrest–induced neuronal pyroptosis,oxidative stress,and mitochondrial damage,whereas knockdown of the liver isoform of phosphofructokinase in bone marrow–derived mesenchymal stem cells inhibited these effects.To conclude,hypoxia-preconditioned bone marrow–derived mesenchymal stem cells offer a promising therapeutic approach for neuronal injury following cardiac arrest,and their beneficial effects are potentially associated with increased expression of the liver isoform of phosphofructokinase following hypoxic preconditioning.展开更多
The aim of this study was to evaluate the comparative effects of hexokinase(HK),phosphofructokinase(PFK)and pyruvate kinase(PK)on glycolysis in postmortem meat.Three specific inhibitors were respectively added to post...The aim of this study was to evaluate the comparative effects of hexokinase(HK),phosphofructokinase(PFK)and pyruvate kinase(PK)on glycolysis in postmortem meat.Three specific inhibitors were respectively added to postmortem lamb to produce a HK inhibition(HKI)group,a PFK inhibition(PFKI)group and a PK inhibition(PKI)group.Meat samples that assigned to these three groups and a control(CON)group were stored at 4◦C for 5 d.The results indicated that the activity of HK,PFK and PK were effectively restrained during storage.PFK activity was significantly correlated with lactate content(P<0.05),but HK and PK activity showed a weak relationship with lactate content(P>0.05).The myofibrillar fragmentation index of the PFKI and PKI groups were significantly lower than that of the CON and HKI groups(P<0.05).The degradation levels of troponin T and desmin were the smallest in the PFKI group.The result of the myoglobin spectrum at 540 nm and 580 nm showed a decreasing level of oxymyoglobin after HK,PFK and PK inhibition.In conclusion,PFK showed a more dominant effect on glycolysis than HK and PK in postmortem muscle,which may provide a targeted strategy for meat quality regulation through glycolysis.展开更多
Phosphofructokinase(PFK)is a key glycolytic rate-limiting enzyme that affects final meat quality through glycolysis regulation.PFK occurs protein phosphorylation and acetylation in postmortem meat,but how they affect ...Phosphofructokinase(PFK)is a key glycolytic rate-limiting enzyme that affects final meat quality through glycolysis regulation.PFK occurs protein phosphorylation and acetylation in postmortem meat,but how they affect PFK activity has not been well studied.Phosphorylation and acetylation of PFK were adjusted in an in vitro system by adding protein kinase A(PKA)and P300/CBP-associated factor(PCAF),respectively.PFK activity significantly decreased after increasing its phosphorylation level by PKA(P<0.05).PCAF increased PFK acetylation level,but did not significantly change its activity(P>0.05).Five PFK variants including wtPFK(wild type),PFK_T704D(T704 phosphorylation),PFK_T704A(T704 dephosphorylation),PFK_K678Q(K678 acetylation)and PFK_K678R(K678 acetylation)were constructed to investigated the effect of T704 and K678 on enzyme activity.Kinetic analysis showed a higher Km,lower Vmax,and lower catalytic efficiency of PFK_T704D on fructose 6-phosphate(F6P),suggesting that T704 phosphorylation decreased PFK activity,possibly by weakening the affinity between F6P and PFK.The changes of PFK variants on ATP were slightly lower than those of PFK variants on F6P.The present study identifies T704 as a possibly vital site for PFK activity regulation and provides theoretical support for the development of meat quality preservation technology.展开更多
Objective： To compare the effects of different methods of anesthesia and analgesia on the activities of phosphofructokinase（PFK）, glucose-6-phosphate dehydrogenase（G-6PD） and aldose reductase（AR） in erythrocyte...Objective： To compare the effects of different methods of anesthesia and analgesia on the activities of phosphofructokinase（PFK）, glucose-6-phosphate dehydrogenase（G-6PD） and aldose reductase（AR） in erythrocytes and levels of plasma glucose and stress hormones in patients undergoing esophagus surgery. Methods： Sixty-two patients scheduled for esophagus surgery were randomly divided into three groups： group Ⅰ （n = 20） receiving only general anesthesia（GA） followed by intravenous patient controlled analgesia（PCA） with fentanyl 15 μg/kg. The other two groups receiving both general anesthesia combined with thoracic epidural anesthesia （GEA） and either intravenous PCA with fentanyl 15 μg/kg （group Ⅱ, n = 21） or thoracic epidural analgesia（TEA） with 0.125% ropivacaine and 0.0002% fentanyl mixture（group Ⅲ, n = 21） after the operation. Venous blood samples were collected for measurements of PFK, G-rPD and AR activities in erythrocytes and plasma glucose, cortisol, epinephrine and norepinephrine before induction （T1）, 60 min following the incision （T2）, 60 min（T3） after operation, on the lst（T4） and 2nd postoperative day（T5）. Results： The activities of PFK decreased（P 〈 0.01 or P = 0.004） and the activities of G-6PD and AR increased（P 〈 0.01） in groups Ⅰ and Ⅱ on T4 compared with those on T1 Between the two groups, the activities of these enzymes in group Ⅱ changed less than those of group Ⅰ （P 〈 0.01 or P 〈 0.05）. These enzymes activities changed slightly in group Ⅲ on T4（P 〉 0.05）. There were significant differences between group Ⅲand the other two groups（P 〈 0.0l or P 〈 0.05）. The levels of plasma glucose increased significantly on T2（P 〈 0.01）, reached peak values on Ta（P 〈 0.01） and fell on T5 in the three groups. Compared to those of groups Ⅰ and Ⅱ, the values of plasma glucose in group Ⅲwere lower on T4 and T5（P 〈 0.05 or P 〈 0.01）. The cortisol concentration in each group increased significantly at T2（P 〈 0.01 or P 〈 0.05）, and remained elevated on T5（P 〈 0.01 or P 〈 0.05）, while on T2 and T3 the cortisol levels＇ of group I were higher than that of groups Ⅱand Ⅲ （P 〈 0.05）. The levels of group Ⅲ were lower than those of the other groups on T4 and T5（P 〈 0.01 or P 〈 0.05）. The levels of epinephrine and norepinephrine were also significantly higher in group Ⅰ than those of the other two groups on T2（P 〈 0.01 or P 〈 0.05）, and their levels in group Ⅰ and Ⅱ were higher than that of group Ⅲ on T4. The patients of the three groups obtained satisfactory pain relief, with all Vidual Analogue Scale（VAS） scores less than 3. VAS scores of group I were much greater 4h after operation. Group m VAS scores were the lowest 24h after operation. However, the number of times patients pressed the bolus switch was higher in group Ⅱ （P 〈 0.01）. Conclusion： Compared with GA and intravenous PCA, general anesthesia combined with thoracic epidural anesthesia and analgesia obtain better pain relief and could markedly alleviate the stress response and improve these erythrocyte glucose metabolism changes after esophagus surgery.展开更多
Impairment of cardiomyocyte mitochondrial function caused by pressure overload is a central event in the development of cardiac hypertrophy(CH).Gentianella acuta Hulten,widely consumed as an herbal tea alternative by ...Impairment of cardiomyocyte mitochondrial function caused by pressure overload is a central event in the development of cardiac hypertrophy(CH).Gentianella acuta Hulten,widely consumed as an herbal tea alternative by the Mongolian and Ewenki hunters,has demonstrated significant protective effects on cardiovascular.However,the bioactive components responsible for the medicinal efficacy of G.acuta,as well as the mechanisms underlying its anti-CH effects,remain incompletely understood.In the present study,we demonstrate that Gen-miR-5,isolated from G.acuta,effectively mitigates angiotensin Ⅱ induced CH.Mechanistically,Gen-miR-5 suppresses the sustained activation of Yes-associated protein(YAP) by downregulating platelet isoform phosphofructokinase,thereby disrupting the YAP/p300/histone deacetylase 6(HDAC6) interaction and subsequently reducing lactylation at the HDAC6 K901 site.Moreover,Gen-miR-5 curtails lactate accumulation in cardiomyocytes,further diminishing lactylation at this site.These processes collectively contribute to the alleviation of mitochondrial oxidative stress,the restoration of mitochondrial membrane potential,and the stabilization of mitochondrial dynamics,thus inhibiting myocardial hypertrophy.These findings highlight the therapeutic potential of miRNAs derived from G.acuta in treating cardiac remodeling diseases and clarify the molecular mechanisms of their cardioprotective effects.展开更多
基金Supported by National Natural Science Foundation Project of China(No.81673979,81473688,81173265)Traditional Chinese Medicine Administration Project of Guangdong Province,China(No.20141070)+3 种基金the Science and Technology Program of Guangdong,China(No.2014A020212672,2014A020210015)the Natural Science Foundation of Guangdong,China(No.2016A030313114,2015A030313333)Scientific Research Cultivation and Innovation of Jinan University Special Fund for Basic Scientific Research of Central University(No.21615464,21615412)the New Century Talent Support Program by the Ministry of Education(No.NCET-13-0827)
文摘OBJECTIVE:To investigate the effect of Ruyanneixiao cream(RYNX) on the expression of hypoxia inducible factor-1α(HIF-1α), hexokinase 2(HK2),phosphofructokinase(PFK), and pyruvate kinase M2(PKM2) mRNA and protein in MCF-10 AT cells and in an animal model of precancerous mammary lesions.METHODS:Following treatment of MCF-10 AT cells with RYNX, tamoxifen(TAM) and YC-1 for 48 h,HIF-1α, HK2, PFK, PKM2 mRNA and protein expression was analyzed.Fifty female SD rats were randomly divided into control, model, TAM, and highand low-dose RYNX groups, with 10 rats in each group.A precancerous mammary lesion model was established for all groups except the control group.High-and low-dose RYNX cream containing TAM was coated on the breasts of animals in the corresponding groups.The rat mammary tissue was removed in the 10 th week and HIF-1α, HK2, PFK,PKM2 mRNA and protein was analyzed.RESULTS:In vitro analyses demonstrated that, compared with the matrix group, HIF-1α, HK2, PFK,PKM2 mRNA and protein expression was significantly decreased in the RYNX group(P < 0.05).Compared with the YC-1 + RYNX group, HK2, PFK,and PKM2 protein expression was significantly reduced in the RYNX group.HIF-1α, HK2, PFK, and PKM2 protein expression was increased significantly in the model group(P < 0.05) compared with the control group, while HIF-1α, HK2, PFK, and PKM2 mRNA and protein expression was significantly decreased in both the high-and low-dose RYNX groups(P < 0.05), with the effect being greater in the high-dose group.CONCLUSION:RYNX can block precancerous breast lesions by decreasing the expression of HK2,PFK, and PKM2 mRNA and protein via inhibition of HIF-1α mRNA and protein overexpression in a dose-dependent manner.
基金supported by the National Natural Science Foundation of China(82072309,22067019,22367023,32160236)the Major Science and Technology Project of Yunnan Province(202402AE090006)+1 种基金the Yunnan Provincial Science and Technology Department-Yunnan University Joint Special Project(202201BF070001-001)the Three-Year Initiative Plan for Strengthening Public Health System Construction in Shanghai(2023–2025)Key Discipline Project(GWV111.1-12)。
文摘The Pomacea canaliculata,also known as apple snail,is an invasive species in China,posing a serious threat to agricultural production and public health.To prevent its rampant infestation,we developed a highly efficient snail-killing lead compound,1-(4-chlorophenyl)-3-(pyridin-3-yl)urea(PBQ).However,the killing mechanism of PBQ remains unclear.To elucidate this,two novel compounds,B-PBQ and J-PBQ,were developed through structural modification of PBQ and were used,through both protein pull-down assay and confocal laser scanning microscopy imaging,to identify and confirm that phosphofructokinase(PFK)in the apple snail is the primary target protein of PBQ.Further studies revealed that PBQ successfully inhibits the activity,content,and expression of PFK in apple snail,disrupting the glycolysis pathway and leading to abnormal energy production.These disruptions impair the snail's tolerance to low oxygen,resulting in its efficient eradication.These findings offer a promising direction for developing next-generation PFK-targeted snail-killing pesticides.Furthermore,the strategy of combining pull-down assays with optical imaging used in this work provides a novel approach for the exploration of mechanisms of other pesticide candidates.
基金supported by grants from the National Key R&D Program of China(2022YFC2503704 to L.-W.D.)the National Natural Science Foundation of China(81988101and 91859205 to H.-Y.W.,82473208 to L.-W.D.,92253306 to Y.-X.T.,and 32270814 and 92359301 to T.-Y.J.)+1 种基金Open Project of State Key Laboratory of Systems Medicine for Cancer(KF2125-93 to L.-W.D.)Shanghai Science and Technology Committee Rising-Star Program(24QA2711800 to T.-Y.J.).
文摘Zinc-dependent histone deacetylases(HDACs)are pivotal enzymes governing the epigenetic modulation of gene expression through chromatin remodeling.The dysregulated expression of HDACs is intricately linked to various pathological conditions,including cancer and inflammation.Histone deacetylase inhibitors(HDACi)have shown therapeutic potential in certain hematologic malignancies.However,the clinical performance of HDACi in solid tumors remains unsatisfactory,and the precise mechanisms of its therapeutic effect in solid tumors has not been fully elucidated.In this study,we identified nucleus-localized PFKL(Liver-type phosphofructokinase),as a key regulator of HDACi efficacy and intracellular epigenetic dynamics.Nuclear PFKL directly binds to classⅠHDACs through interacting with zinc-binding sites,thereby inhibiting HDAC enzymatic activity and promoting intracellular histone acetylation.In addition,the Thr562 residue within PFKL enhances the chelation effect between the zinc-binding group(ZBG)of the HDACi romidepsin and the zinc within the HDACs,further promoting drug efficacy.Based on the mechanism of PFKL facilitates the efficacy of romidepsin,we developed a therapeutic peptide,PFKL-552-572-R8,which significantly enhances the antitumor effect of romidepsin both in vitro and in vivo.Our findings reveal that spatiotemporal regulation confers a moonlight function to PFKL as an endogenous HDAC inhibitor to maintain the stability of epigenetic modifications and highlight PFKL as a promising therapeutic target for enhancing the clinical utility of HDACi in solid tumors.
基金supported by grants from the National Natural Science Foundation of China(32122080,31972375,31902049)National Key Research and Development Program of China(2018YFD1000200)Shandong Province(ZR2020YQ25)。
文摘Sugars are involved in plant growth,fruit quality,and signaling perception.Therefore,understanding the mechanisms involved in soluble sugar accumulation is essential to understand fruit development.Here,we report that Md PFPβ,a pyrophosphatedependent phosphofructokinase gene,regulates soluble sugar accumulation by enhancing the photosynthetic performance and sugar-metabolizing enzyme activities in apple(Malus domestica Borkh.).Biochemical analysis revealed that a basic helix-loop-helix(b HLH)transcription factor,Mdb HLH3,binds to the Md PFPβpromoter and activates its expression,thus promoting soluble sugar accumulation in apple fruit.In addition,Md PFPβoverexpression in tomato influenced photosynthesis and carbon metabolism in the plant.Furthermore,we determined that Mdb HLH3 increases photosynthetic rates and soluble sugar accumulation in apple by activating Md PFPβexpression.Our results thus shed light on the mechanism of soluble sugar accumulation in apple leaves and fruit:Mdb HLH3 regulates soluble sugar accumulation by activating Md PFPβgene expression and coordinating carbohydrate allocation.
基金supported by the Natural Science Fund of Fujian Province,No.2020J011058(to JK)the Project of Fujian Provincial Hospital for High-level Hospital Construction,No.2020HSJJ12(to JK)+1 种基金the Fujian Provincial Finance Department Special Fund,No.(2021)848(to FC)the Fujian Provincial Major Scientific and Technological Special Projects on Health,No.2022ZD01008(to FC).
文摘Cardiac arrest can lead to severe neurological impairment as a result of inflammation,mitochondrial dysfunction,and post-cardiopulmonary resuscitation neurological damage.Hypoxic preconditioning has been shown to improve migration and survival of bone marrow–derived mesenchymal stem cells and reduce pyroptosis after cardiac arrest,but the specific mechanisms by which hypoxia-preconditioned bone marrow–derived mesenchymal stem cells protect against brain injury after cardiac arrest are unknown.To this end,we established an in vitro co-culture model of bone marrow–derived mesenchymal stem cells and oxygen–glucose deprived primary neurons and found that hypoxic preconditioning enhanced the protective effect of bone marrow stromal stem cells against neuronal pyroptosis,possibly through inhibition of the MAPK and nuclear factor κB pathways.Subsequently,we transplanted hypoxia-preconditioned bone marrow–derived mesenchymal stem cells into the lateral ventricle after the return of spontaneous circulation in an 8-minute cardiac arrest rat model induced by asphyxia.The results showed that hypoxia-preconditioned bone marrow–derived mesenchymal stem cells significantly reduced cardiac arrest–induced neuronal pyroptosis,oxidative stress,and mitochondrial damage,whereas knockdown of the liver isoform of phosphofructokinase in bone marrow–derived mesenchymal stem cells inhibited these effects.To conclude,hypoxia-preconditioned bone marrow–derived mesenchymal stem cells offer a promising therapeutic approach for neuronal injury following cardiac arrest,and their beneficial effects are potentially associated with increased expression of the liver isoform of phosphofructokinase following hypoxic preconditioning.
基金supported by the National Natural Science Foundation of China(32372263).
文摘The aim of this study was to evaluate the comparative effects of hexokinase(HK),phosphofructokinase(PFK)and pyruvate kinase(PK)on glycolysis in postmortem meat.Three specific inhibitors were respectively added to postmortem lamb to produce a HK inhibition(HKI)group,a PFK inhibition(PFKI)group and a PK inhibition(PKI)group.Meat samples that assigned to these three groups and a control(CON)group were stored at 4◦C for 5 d.The results indicated that the activity of HK,PFK and PK were effectively restrained during storage.PFK activity was significantly correlated with lactate content(P<0.05),but HK and PK activity showed a weak relationship with lactate content(P>0.05).The myofibrillar fragmentation index of the PFKI and PKI groups were significantly lower than that of the CON and HKI groups(P<0.05).The degradation levels of troponin T and desmin were the smallest in the PFKI group.The result of the myoglobin spectrum at 540 nm and 580 nm showed a decreasing level of oxymyoglobin after HK,PFK and PK inhibition.In conclusion,PFK showed a more dominant effect on glycolysis than HK and PK in postmortem muscle,which may provide a targeted strategy for meat quality regulation through glycolysis.
基金supported by the National Natural Science Foundation of China(32030086,32072245)Agricultural Science and Technology Innovation Program(CAAS-ASTIP-2023-IFST-03)in China.
文摘Phosphofructokinase(PFK)is a key glycolytic rate-limiting enzyme that affects final meat quality through glycolysis regulation.PFK occurs protein phosphorylation and acetylation in postmortem meat,but how they affect PFK activity has not been well studied.Phosphorylation and acetylation of PFK were adjusted in an in vitro system by adding protein kinase A(PKA)and P300/CBP-associated factor(PCAF),respectively.PFK activity significantly decreased after increasing its phosphorylation level by PKA(P<0.05).PCAF increased PFK acetylation level,but did not significantly change its activity(P>0.05).Five PFK variants including wtPFK(wild type),PFK_T704D(T704 phosphorylation),PFK_T704A(T704 dephosphorylation),PFK_K678Q(K678 acetylation)and PFK_K678R(K678 acetylation)were constructed to investigated the effect of T704 and K678 on enzyme activity.Kinetic analysis showed a higher Km,lower Vmax,and lower catalytic efficiency of PFK_T704D on fructose 6-phosphate(F6P),suggesting that T704 phosphorylation decreased PFK activity,possibly by weakening the affinity between F6P and PFK.The changes of PFK variants on ATP were slightly lower than those of PFK variants on F6P.The present study identifies T704 as a possibly vital site for PFK activity regulation and provides theoretical support for the development of meat quality preservation technology.
基金supported by Jiangsu Province Department of health Fund(No.H200705)
文摘Objective： To compare the effects of different methods of anesthesia and analgesia on the activities of phosphofructokinase（PFK）, glucose-6-phosphate dehydrogenase（G-6PD） and aldose reductase（AR） in erythrocytes and levels of plasma glucose and stress hormones in patients undergoing esophagus surgery. Methods： Sixty-two patients scheduled for esophagus surgery were randomly divided into three groups： group Ⅰ （n = 20） receiving only general anesthesia（GA） followed by intravenous patient controlled analgesia（PCA） with fentanyl 15 μg/kg. The other two groups receiving both general anesthesia combined with thoracic epidural anesthesia （GEA） and either intravenous PCA with fentanyl 15 μg/kg （group Ⅱ, n = 21） or thoracic epidural analgesia（TEA） with 0.125% ropivacaine and 0.0002% fentanyl mixture（group Ⅲ, n = 21） after the operation. Venous blood samples were collected for measurements of PFK, G-rPD and AR activities in erythrocytes and plasma glucose, cortisol, epinephrine and norepinephrine before induction （T1）, 60 min following the incision （T2）, 60 min（T3） after operation, on the lst（T4） and 2nd postoperative day（T5）. Results： The activities of PFK decreased（P 〈 0.01 or P = 0.004） and the activities of G-6PD and AR increased（P 〈 0.01） in groups Ⅰ and Ⅱ on T4 compared with those on T1 Between the two groups, the activities of these enzymes in group Ⅱ changed less than those of group Ⅰ （P 〈 0.01 or P 〈 0.05）. These enzymes activities changed slightly in group Ⅲ on T4（P 〉 0.05）. There were significant differences between group Ⅲand the other two groups（P 〈 0.0l or P 〈 0.05）. The levels of plasma glucose increased significantly on T2（P 〈 0.01）, reached peak values on Ta（P 〈 0.01） and fell on T5 in the three groups. Compared to those of groups Ⅰ and Ⅱ, the values of plasma glucose in group Ⅲwere lower on T4 and T5（P 〈 0.05 or P 〈 0.01）. The cortisol concentration in each group increased significantly at T2（P 〈 0.01 or P 〈 0.05）, and remained elevated on T5（P 〈 0.01 or P 〈 0.05）, while on T2 and T3 the cortisol levels＇ of group I were higher than that of groups Ⅱand Ⅲ （P 〈 0.05）. The levels of group Ⅲ were lower than those of the other groups on T4 and T5（P 〈 0.01 or P 〈 0.05）. The levels of epinephrine and norepinephrine were also significantly higher in group Ⅰ than those of the other two groups on T2（P 〈 0.01 or P 〈 0.05）, and their levels in group Ⅰ and Ⅱ were higher than that of group Ⅲ on T4. The patients of the three groups obtained satisfactory pain relief, with all Vidual Analogue Scale（VAS） scores less than 3. VAS scores of group I were much greater 4h after operation. Group m VAS scores were the lowest 24h after operation. However, the number of times patients pressed the bolus switch was higher in group Ⅱ （P 〈 0.01）. Conclusion： Compared with GA and intravenous PCA, general anesthesia combined with thoracic epidural anesthesia and analgesia obtain better pain relief and could markedly alleviate the stress response and improve these erythrocyte glucose metabolism changes after esophagus surgery.
基金supported by the National Natural Science Foundation of China(82174080)the S&T Program of Hebei(20372501D)+1 种基金the Hebei Natural Science Foundation(H2021423069,H2022423333)the Science Research Project of Hebei Education Department(BJ2021031)。
文摘Impairment of cardiomyocyte mitochondrial function caused by pressure overload is a central event in the development of cardiac hypertrophy(CH).Gentianella acuta Hulten,widely consumed as an herbal tea alternative by the Mongolian and Ewenki hunters,has demonstrated significant protective effects on cardiovascular.However,the bioactive components responsible for the medicinal efficacy of G.acuta,as well as the mechanisms underlying its anti-CH effects,remain incompletely understood.In the present study,we demonstrate that Gen-miR-5,isolated from G.acuta,effectively mitigates angiotensin Ⅱ induced CH.Mechanistically,Gen-miR-5 suppresses the sustained activation of Yes-associated protein(YAP) by downregulating platelet isoform phosphofructokinase,thereby disrupting the YAP/p300/histone deacetylase 6(HDAC6) interaction and subsequently reducing lactylation at the HDAC6 K901 site.Moreover,Gen-miR-5 curtails lactate accumulation in cardiomyocytes,further diminishing lactylation at this site.These processes collectively contribute to the alleviation of mitochondrial oxidative stress,the restoration of mitochondrial membrane potential,and the stabilization of mitochondrial dynamics,thus inhibiting myocardial hypertrophy.These findings highlight the therapeutic potential of miRNAs derived from G.acuta in treating cardiac remodeling diseases and clarify the molecular mechanisms of their cardioprotective effects.
