Platelets are indispensable for physiological hemostasis and pathological thrombus formation,and platelet adhesion to endothelial collagen is a critical initial step in thrombus formation,often overlooked in current a...Platelets are indispensable for physiological hemostasis and pathological thrombus formation,and platelet adhesion to endothelial collagen is a critical initial step in thrombus formation,often overlooked in current antiplatelet therapies.This study aims to elucidate how ginsenoside CK enhances hemodynamic circulation,alleviates stasis,and proposes therapeutic mechanisms.Inspired by the effects on improving microcirculatory disturbances in an acute soft tissue injury model,CK was identified as a PHD2 inhibitor,effectively suppressing platelet adhesion to collagen.It was proposed that targeting PHD2 regulates collagen hydroxylation modification,thereby influencing the formation of its three-dimensional structure,reducing the binding affinity between VWF and collagen,and ultimately suppressing thrombotic events.The efficacy of this mechanism was subsequently confirmed through a mouse DIC model,demonstrating the feasibility of CK in alleviating circulatory disorders.It is worth noting that when Phd2 was knocked down in mice's lungs,pulmonary embolism was significantly reduced.Additionally,PHD2 inhibitors approved for other diseases have exhibited similar anti-thrombotic effects.Moreover,when PHD2 inhibitors were combined with aspirin,they more effectively inhibited arterial thrombosis in rats.The findings offer valuable insights into potential targets for developing antiplatelet drugs or expanding therapeutic applications for existing PHD2 inhibitors in treating thrombotic diseases.展开更多
Osteoporosis(OP),a systemic and chronic bone disease,is distinguished by low bone mass and destruction of bone microarchitecture.Ginsenoside Compound-K(CK),one of the metabolites of ginsenoside Rb1,has anti-aging,anti...Osteoporosis(OP),a systemic and chronic bone disease,is distinguished by low bone mass and destruction of bone microarchitecture.Ginsenoside Compound-K(CK),one of the metabolites of ginsenoside Rb1,has anti-aging,anti-inflammatory,anti-cancer,and hypolipidemic activities.We have demonstrated CK could promote osteogenesis and fracture healing in our previous study.However,the contribution of CK to osteoporosis has not been examined.In the present study,we investigated the effect of CK on osteoclastogenesis and ovariectomy(OVX)-induced osteoporosis.The results showed that CK inhibited receptor activator for nuclear factor-κB ligand(RANKL)-mediated osteoclast differentiation and reactive oxygen species(ROS)activity by inhibiting the phosphorylation of NF-κB p65 and oxidative stress in RAW264.7 cells.In addition,we also demonstrated that CK could inhibit bone resorption using bone marrow-derived macrophages.Furthermore,we demonstrated that CK attenuated bone loss by suppressing the activity of osteoclast and alleviating oxidative stress in vivo.Taken together,these results showed CK could inhibit osteoclastogenesis and prevent OVX-induced bone loss by inhibiting NF-κB signaling pathway.展开更多
Glycosidases play a pivotal role in the biocatalytic production of high-value glycosides,but their industrial applications are often limited by poor thermo-pH tolerance and incompatible reaction conditions in multienz...Glycosidases play a pivotal role in the biocatalytic production of high-value glycosides,but their industrial applications are often limited by poor thermo-pH tolerance and incompatible reaction conditions in multienzyme systems.To address these universal challenges,we developed a rational N-terminal truncation strategy usingβ-glucosidase B6 as a model.By removing 33 hydrophilic residues from the terminus,the engineered enzyme(33aa)exhibited significantly improved properties:a 10◦C higher optimal temperature(50◦C),enhanced acid tolerance(>50%activity at pH 5.0),and stronger substrate affinity(K_(m)=0.8517 mmol/L).Structural analysis revealed that the truncation reduced solvent-accessible surface area(SASA)and increasedα-helix density,explaining its superior stability.To demonstrate industrial application potential,we paired 33aa with G4 glucosidase-a thermophilic enzyme with complementary regioselectivity-creating an efficient cascade system.The optimized conditions(57.5◦C,pH 6.0,5:1 activity ratio)achieved a 3.32-fold higher product yield compared to single-enzyme systems in ginsenoside C-K production.This study establishes N-terminal truncation as a generalizable protein engineering approach to synchronize reaction conditions and enhance catalytic efficiency in glycosidase-based bioconversions,offering a versatile solution for industrial glycoside production.展开更多
基金supported by National Natural Science Foundation of China,grant number:82274184.
文摘Platelets are indispensable for physiological hemostasis and pathological thrombus formation,and platelet adhesion to endothelial collagen is a critical initial step in thrombus formation,often overlooked in current antiplatelet therapies.This study aims to elucidate how ginsenoside CK enhances hemodynamic circulation,alleviates stasis,and proposes therapeutic mechanisms.Inspired by the effects on improving microcirculatory disturbances in an acute soft tissue injury model,CK was identified as a PHD2 inhibitor,effectively suppressing platelet adhesion to collagen.It was proposed that targeting PHD2 regulates collagen hydroxylation modification,thereby influencing the formation of its three-dimensional structure,reducing the binding affinity between VWF and collagen,and ultimately suppressing thrombotic events.The efficacy of this mechanism was subsequently confirmed through a mouse DIC model,demonstrating the feasibility of CK in alleviating circulatory disorders.It is worth noting that when Phd2 was knocked down in mice's lungs,pulmonary embolism was significantly reduced.Additionally,PHD2 inhibitors approved for other diseases have exhibited similar anti-thrombotic effects.Moreover,when PHD2 inhibitors were combined with aspirin,they more effectively inhibited arterial thrombosis in rats.The findings offer valuable insights into potential targets for developing antiplatelet drugs or expanding therapeutic applications for existing PHD2 inhibitors in treating thrombotic diseases.
基金the grant from National Natural Science Foundation of China(81871778)Guangdong Provincial Science and Technology Collaborative Innovation Center for Sport Science(2019B110210004)the key project of Sport Research Foundation of Guangdong Province(GDSS2022M005).
文摘Osteoporosis(OP),a systemic and chronic bone disease,is distinguished by low bone mass and destruction of bone microarchitecture.Ginsenoside Compound-K(CK),one of the metabolites of ginsenoside Rb1,has anti-aging,anti-inflammatory,anti-cancer,and hypolipidemic activities.We have demonstrated CK could promote osteogenesis and fracture healing in our previous study.However,the contribution of CK to osteoporosis has not been examined.In the present study,we investigated the effect of CK on osteoclastogenesis and ovariectomy(OVX)-induced osteoporosis.The results showed that CK inhibited receptor activator for nuclear factor-κB ligand(RANKL)-mediated osteoclast differentiation and reactive oxygen species(ROS)activity by inhibiting the phosphorylation of NF-κB p65 and oxidative stress in RAW264.7 cells.In addition,we also demonstrated that CK could inhibit bone resorption using bone marrow-derived macrophages.Furthermore,we demonstrated that CK attenuated bone loss by suppressing the activity of osteoclast and alleviating oxidative stress in vivo.Taken together,these results showed CK could inhibit osteoclastogenesis and prevent OVX-induced bone loss by inhibiting NF-κB signaling pathway.
基金funded by Shandong Provincial Technological Innovation Guidance Plan(YDZX2024106)the National Key Research and Development Program of China(2021YFC2102800)the National Natural Science Foundation of China(22078014).
文摘Glycosidases play a pivotal role in the biocatalytic production of high-value glycosides,but their industrial applications are often limited by poor thermo-pH tolerance and incompatible reaction conditions in multienzyme systems.To address these universal challenges,we developed a rational N-terminal truncation strategy usingβ-glucosidase B6 as a model.By removing 33 hydrophilic residues from the terminus,the engineered enzyme(33aa)exhibited significantly improved properties:a 10◦C higher optimal temperature(50◦C),enhanced acid tolerance(>50%activity at pH 5.0),and stronger substrate affinity(K_(m)=0.8517 mmol/L).Structural analysis revealed that the truncation reduced solvent-accessible surface area(SASA)and increasedα-helix density,explaining its superior stability.To demonstrate industrial application potential,we paired 33aa with G4 glucosidase-a thermophilic enzyme with complementary regioselectivity-creating an efficient cascade system.The optimized conditions(57.5◦C,pH 6.0,5:1 activity ratio)achieved a 3.32-fold higher product yield compared to single-enzyme systems in ginsenoside C-K production.This study establishes N-terminal truncation as a generalizable protein engineering approach to synchronize reaction conditions and enhance catalytic efficiency in glycosidase-based bioconversions,offering a versatile solution for industrial glycoside production.
