Alzheimer’s disease is a neurodegenerative disease induced by multiple interconnected mechanisms.Peptide drug candidates with multi-modal efficacy generated from fusion strategy are suitable for addressing multi-face...Alzheimer’s disease is a neurodegenerative disease induced by multiple interconnected mechanisms.Peptide drug candidates with multi-modal efficacy generated from fusion strategy are suitable for addressing multi-facet pathology.However,clinical translation of peptide drugs is greatly hampered by their low permeability into brain.Herein,a hybrid peptide HNSS is generated by merging two therapeutic peptides(SS31 and S-14 G Humanin(HNG)),using a different approach from the classical shuttle-therapeutic peptide conjugate design.HNSS demonstrated increased bio-permeability,with a 2-fold improvement in brain distribution over HNG,thanks to its structure mimicking the design of signal peptide-derived cell-penetrating peptides.HNSS efficiently alleviated mitochondrial dysfunction through the combined effects of mitochondrial targeting,ROS scavenging and p-STAT3 activation.Meanwhile,HNSS with increased Aβaffinity greatly inhibited Aβoligomerization/fibrillation,and interrupted Aβinteraction with neuron/microglia by reducing neuronal mitochondrial Aβdeposition and promoting microglial phagocytosis of Aβ.In3×Tg-AD transgenic mice,HNSS treatment efficiently inhibited brain neuron loss and improved the cognitive performance.This work validates the rational fusion design-based strategy for bio-permeability improvement and efficacy amplification,providing a paradigm for developing therapeutic peptide candidates against neurodegenerative disease.展开更多
To optimize the conditions for the establishment of two-dimensional gel electrophoresis(2-DE)of soybean protein isolate(SPI),we investigated Ampholine mixture,anodic and cathodic electrolytes,loading amount of sample,...To optimize the conditions for the establishment of two-dimensional gel electrophoresis(2-DE)of soybean protein isolate(SPI),we investigated Ampholine mixture,anodic and cathodic electrolytes,loading amount of sample,acrylamide concentration,p H gradient and gel staining method in twodimensional gel electrophoresis to optimize the protein imaging conditions in two-dimensional gel.The results of mixed-level design experiments showed that Ampholine,loading amount and gel staining method had significant effect(P<0.05)on 2-DE of SPI.The optimal conditions were Ampholine mixture(pH 3–10+pH 5–7 or pH 4–6+pH 5–7),loading amount of 2 mg sample and silver staining.Although the acrylamide concentration of the gel,the p H gradient,the anodic and cathodic electrolyte solutions had significant statistical effects on the protein separation degree,the complexity of the protein composition and the visibility of the gel images were more inclined to the 12%gel,the 3–10 pH gradient and the H3PO4/NaOH electrolyte.According to the established conditions,the hydrolyzed products of SPI emulsion were determined by 2-DE,and the dynamic changes of protein in the process of enzymatic hydrolysis were described.展开更多
Objective:Primary tumor treatment through surgical resection and adjuvant therapy has been extensively studied,but there is a lack of effective strategies and drugs for the treatment of tumor metastases.Here,we descri...Objective:Primary tumor treatment through surgical resection and adjuvant therapy has been extensively studied,but there is a lack of effective strategies and drugs for the treatment of tumor metastases.Here,we describe a functional product based on a combination of compounds,which can be used as an adjuvant therapy and has well-known mechanisms for inhibiting cancer metastases,improving anti-cancer treatment,and enhancing immunity and antioxidant capacity.Our designed combination,named MVBL,consists of four inexpensive compounds:L-selenium-methylselenocysteine(MSC),D-α-tocopheryl succinic acid(VES),β-carotene(β-Ca),and L-lysine(Lys).Methods:The effects of MVBL on cell viability,cell cycle,cell apoptosis,cell migration,cell invasion,reactive oxygen species(ROS),and paclitaxel(PTX)-combined treatment were studied in vitro.The inhibition of tumor metastasis,antioxidation,and immune enhancement capacity of MVBL were determined in vivo.Results:MVBL exhibited higher toxicity to tumor cells than to normal cells.It did not significantly affect the cell cycle of cancer cells,but increased their apoptosis.Wound healing,adhesion,and transwell assays showed that MVBL significantly inhibited tumor cell migration,adhesion,and invasion.MVBL sensitized MDA-MB-231 breast cancer cells to PTX,indicating that it can be used as an adjuvant to enhance the therapeutic effect of chemotherapy drugs.In mice,experimental data showed that MVBL inhibited tumor metastasis,prolonged their survival time,and enhanced their antioxidant capacity and immune function.Conclusions:This study revealed the roles of MVBL in improving immunity and antioxidation,preventing tumor growth,and inhibiting metastasis in vitro and in vivo.MVBL may be used as an adjuvant drug in cancer therapy for improving the survival and quality of life of cancer patients.展开更多
Intraneuronal dysproteostasis and extraneuronal microenvironmental abnormalities in Alzheimer’s disease(AD)collectively culminate in neuronal deterioration.In the context of AD,autophagy dysfunction,a multi-link obst...Intraneuronal dysproteostasis and extraneuronal microenvironmental abnormalities in Alzheimer’s disease(AD)collectively culminate in neuronal deterioration.In the context of AD,autophagy dysfunction,a multi-link obstacle involving autophagy downregulation and lysosome defects in neurons/microglia is highly implicated in intra/extraneuronal pathological processes.Therefore,multidimensional autophagy regulation strategies co-manipulating“autophagy induction”and“lysosome degradation”in dual targets(neuron and microglia)are more reliable for AD treatment.Accordingly,we designed an RP-1 peptide-modified reactive oxygen species(ROS)-responsive micelles(RT-NM)loading rapamycin or gypenoside XVII.Guided by RP-1 peptide,the ligand of receptor for advanced glycation end products(RAGE),RT-NM efficiently targeted neurons and microglia in AD-affected region.This nanocombination therapy activated the whole autophagy-lysosome pathway by autophagy induction(rapamycin)and lysosome improvement(gypenoside XVII),thus enhancing autophagic degradation of neurotoxic aggregates and inflammasomes,and promoting Aβ phagocytosis.Resultantly,it decreased aberrant protein burden,alleviated neuroinflammation,and eventually ameliorated memory defects in 3×Tg-AD transgenic mice.Our research developed a multidimensional autophagy nano-regulator to boost the efficacy of autophagy-centered AD therapy.展开更多
Compromised autophagy and defective lysosomal clearance significantly contribute to impaired neuronal proteostasis,which represents a hallmark of Alzheimer’s disease(AD)and other age-related neurodegenerative disorde...Compromised autophagy and defective lysosomal clearance significantly contribute to impaired neuronal proteostasis,which represents a hallmark of Alzheimer’s disease(AD)and other age-related neurodegenerative disorders.Growing evidence has implicated that modulating autophagic flux,instead of inducing autophagosome formation alone,would be more reliable to rescue neuronal proteostasis.Concurrently,selectively enhancing drug concentrations in the leision areas,instead of the whole brain,will maximize therapeutic efficacy while reduing non-selective autophagy induction.Herein,we design a ROS-responsive targeted micelle system(TT-NM/Rapa)to enhance the delivery efficiency of rapamycin to neurons in AD lesions guided by the fusion peptide TPL,and facilitate its intracellular release via ROS-mediated disassembly of micelles,thereby maximizing autophagic flux modulating efficacy of rapamycin in neurons.Consequently,it promotes the efficient clearance of intracellular neurotoxic proteins,β-amyloid and hyperphosphorylated tau proteins,and ameliorates memory defects and neuronal damage in 3×Tg-AD transgenic mice.Our studies demonstrate a promising strategy to restore autophagic flux and improve neuronal proteostasis by rationally-engineered nano-systems for delaying the progression of AD.展开更多
基金supported by National Natural Science Foundation of China(82273868 and 82073780)Shanghai Municipal Natural Science Foundation(19ZR1406200).
文摘Alzheimer’s disease is a neurodegenerative disease induced by multiple interconnected mechanisms.Peptide drug candidates with multi-modal efficacy generated from fusion strategy are suitable for addressing multi-facet pathology.However,clinical translation of peptide drugs is greatly hampered by their low permeability into brain.Herein,a hybrid peptide HNSS is generated by merging two therapeutic peptides(SS31 and S-14 G Humanin(HNG)),using a different approach from the classical shuttle-therapeutic peptide conjugate design.HNSS demonstrated increased bio-permeability,with a 2-fold improvement in brain distribution over HNG,thanks to its structure mimicking the design of signal peptide-derived cell-penetrating peptides.HNSS efficiently alleviated mitochondrial dysfunction through the combined effects of mitochondrial targeting,ROS scavenging and p-STAT3 activation.Meanwhile,HNSS with increased Aβaffinity greatly inhibited Aβoligomerization/fibrillation,and interrupted Aβinteraction with neuron/microglia by reducing neuronal mitochondrial Aβdeposition and promoting microglial phagocytosis of Aβ.In3×Tg-AD transgenic mice,HNSS treatment efficiently inhibited brain neuron loss and improved the cognitive performance.This work validates the rational fusion design-based strategy for bio-permeability improvement and efficacy amplification,providing a paradigm for developing therapeutic peptide candidates against neurodegenerative disease.
基金the National Natural Science Foundation of China(No.31371782)the Key Project of Science and Technology Research of Henan Education Department(No.14A550007)the Basic Research Project of Henan University of Technology(No.171157)。
文摘To optimize the conditions for the establishment of two-dimensional gel electrophoresis(2-DE)of soybean protein isolate(SPI),we investigated Ampholine mixture,anodic and cathodic electrolytes,loading amount of sample,acrylamide concentration,p H gradient and gel staining method in twodimensional gel electrophoresis to optimize the protein imaging conditions in two-dimensional gel.The results of mixed-level design experiments showed that Ampholine,loading amount and gel staining method had significant effect(P<0.05)on 2-DE of SPI.The optimal conditions were Ampholine mixture(pH 3–10+pH 5–7 or pH 4–6+pH 5–7),loading amount of 2 mg sample and silver staining.Although the acrylamide concentration of the gel,the p H gradient,the anodic and cathodic electrolyte solutions had significant statistical effects on the protein separation degree,the complexity of the protein composition and the visibility of the gel images were more inclined to the 12%gel,the 3–10 pH gradient and the H3PO4/NaOH electrolyte.According to the established conditions,the hydrolyzed products of SPI emulsion were determined by 2-DE,and the dynamic changes of protein in the process of enzymatic hydrolysis were described.
基金the National Natural Science Foundation of China(Nos.81961138017 and 81703555)the Natural Science Foundation of Fujian Province(Nos.2021J011046 and 2020J01859)+4 种基金the Young and Middle-aged Teacher Education Research Project of Fujian Province(No.JAT200412)the Science and Technology Planning Projects of Fuzhou(No.2021S094)the Science and Technology Planning Projects of Fuzhou Institute of Oceanography(No.2021F08)the Belt&Road Program(No.KXPT-2021-3,CAST)the Ministry of Science and Technology of China(No.2015CB931804)。
文摘Objective:Primary tumor treatment through surgical resection and adjuvant therapy has been extensively studied,but there is a lack of effective strategies and drugs for the treatment of tumor metastases.Here,we describe a functional product based on a combination of compounds,which can be used as an adjuvant therapy and has well-known mechanisms for inhibiting cancer metastases,improving anti-cancer treatment,and enhancing immunity and antioxidant capacity.Our designed combination,named MVBL,consists of four inexpensive compounds:L-selenium-methylselenocysteine(MSC),D-α-tocopheryl succinic acid(VES),β-carotene(β-Ca),and L-lysine(Lys).Methods:The effects of MVBL on cell viability,cell cycle,cell apoptosis,cell migration,cell invasion,reactive oxygen species(ROS),and paclitaxel(PTX)-combined treatment were studied in vitro.The inhibition of tumor metastasis,antioxidation,and immune enhancement capacity of MVBL were determined in vivo.Results:MVBL exhibited higher toxicity to tumor cells than to normal cells.It did not significantly affect the cell cycle of cancer cells,but increased their apoptosis.Wound healing,adhesion,and transwell assays showed that MVBL significantly inhibited tumor cell migration,adhesion,and invasion.MVBL sensitized MDA-MB-231 breast cancer cells to PTX,indicating that it can be used as an adjuvant to enhance the therapeutic effect of chemotherapy drugs.In mice,experimental data showed that MVBL inhibited tumor metastasis,prolonged their survival time,and enhanced their antioxidant capacity and immune function.Conclusions:This study revealed the roles of MVBL in improving immunity and antioxidation,preventing tumor growth,and inhibiting metastasis in vitro and in vivo.MVBL may be used as an adjuvant drug in cancer therapy for improving the survival and quality of life of cancer patients.
基金supported by National Natural Science Foundation of China(Nos.82073780 and 82273868,China)Shanghai Municipal Natural Science Foundation(No.19ZR1406200,China)。
文摘Intraneuronal dysproteostasis and extraneuronal microenvironmental abnormalities in Alzheimer’s disease(AD)collectively culminate in neuronal deterioration.In the context of AD,autophagy dysfunction,a multi-link obstacle involving autophagy downregulation and lysosome defects in neurons/microglia is highly implicated in intra/extraneuronal pathological processes.Therefore,multidimensional autophagy regulation strategies co-manipulating“autophagy induction”and“lysosome degradation”in dual targets(neuron and microglia)are more reliable for AD treatment.Accordingly,we designed an RP-1 peptide-modified reactive oxygen species(ROS)-responsive micelles(RT-NM)loading rapamycin or gypenoside XVII.Guided by RP-1 peptide,the ligand of receptor for advanced glycation end products(RAGE),RT-NM efficiently targeted neurons and microglia in AD-affected region.This nanocombination therapy activated the whole autophagy-lysosome pathway by autophagy induction(rapamycin)and lysosome improvement(gypenoside XVII),thus enhancing autophagic degradation of neurotoxic aggregates and inflammasomes,and promoting Aβ phagocytosis.Resultantly,it decreased aberrant protein burden,alleviated neuroinflammation,and eventually ameliorated memory defects in 3×Tg-AD transgenic mice.Our research developed a multidimensional autophagy nano-regulator to boost the efficacy of autophagy-centered AD therapy.
基金supported by National Natural Science Foundation of China(No.82073780 and 81690263)Shanghai Municipal Natural Science Foundation(No.19ZR140620).
文摘Compromised autophagy and defective lysosomal clearance significantly contribute to impaired neuronal proteostasis,which represents a hallmark of Alzheimer’s disease(AD)and other age-related neurodegenerative disorders.Growing evidence has implicated that modulating autophagic flux,instead of inducing autophagosome formation alone,would be more reliable to rescue neuronal proteostasis.Concurrently,selectively enhancing drug concentrations in the leision areas,instead of the whole brain,will maximize therapeutic efficacy while reduing non-selective autophagy induction.Herein,we design a ROS-responsive targeted micelle system(TT-NM/Rapa)to enhance the delivery efficiency of rapamycin to neurons in AD lesions guided by the fusion peptide TPL,and facilitate its intracellular release via ROS-mediated disassembly of micelles,thereby maximizing autophagic flux modulating efficacy of rapamycin in neurons.Consequently,it promotes the efficient clearance of intracellular neurotoxic proteins,β-amyloid and hyperphosphorylated tau proteins,and ameliorates memory defects and neuronal damage in 3×Tg-AD transgenic mice.Our studies demonstrate a promising strategy to restore autophagic flux and improve neuronal proteostasis by rationally-engineered nano-systems for delaying the progression of AD.
