Background Exosomes are crucial mediators of intercellular communication.As a key component of milk,milkderived exosomes are abundant in genetic cargo,particularly micro RNAs(mi RNAs),indicating their potential role i...Background Exosomes are crucial mediators of intercellular communication.As a key component of milk,milkderived exosomes are abundant in genetic cargo,particularly micro RNAs(mi RNAs),indicating their potential role in regulating mammary gland physiology.Therefore,this study aimed to investigate the specificity of mi RNAs in milkderived exosomes and their regulatory roles in lipid synthesis in bovine mammary epithelial cells(BMECs).Results Based on 17,838 DHI records showing a significantly higher milk fat percentage(MFP)in late lactation(4.24%±1.07%),10 high-(5.96%±0.26%,HMF)and 10 low-MFP(1.68%±0.23%,LMF)cows were selected during this stage for milk-derived exosome isolation and mi RNA profiling.Exosomes isolated via differential ultracentrifugation were verified as 50-150 nm vesicles expressing CD9,CD81,and TSG101.mi RNA sequencing identified 1,320 differentially expressed mi RNAs(496 upregulated and 824 downregulated)between the HMF_EXO and LMF_EXO groups.Uptake assays confirmed that BMECs internalized these exosomes,and q RT-PCR validation showed that mi R-423-5p and mi R-125b were significantly upregulated and downregulated in HMF_EXO-and LMF_EXO-treated BMECs,respectively.Functionally,exosomal mi R-423-5p promoted intracellular lipid accumulation and TG synthesis in BMECs by targeting APOA5,whereas mi R-125b inhibited lipolysis and fatty acid oxidation by repressing SLC27A1.Conclusions This study demonstrates that milk-derived exosomal mi RNAs represent a novel mechanism for regulating milk fat synthesis.Specifically,mi R-423-5p and mi R-125b directly modulated lipid metabolism in BMECs via the mi R-423-5p/APOA5 and mi R-125b/SLC27A1 pathways.These findings provide new insights into the molecular regulation of milk fat synthesis and highlight the importance of exosome-mediated intercellular communication in the lactating mammary gland.展开更多
Milk-derived extracellular vesicles(EVs)are promising for oral drug delivery,yet different loading methods exhibit distinct impacts on drug encapsulation and membrane integrity.This study demonstrated that sonication ...Milk-derived extracellular vesicles(EVs)are promising for oral drug delivery,yet different loading methods exhibit distinct impacts on drug encapsulation and membrane integrity.This study demonstrated that sonication method achieved high drug encapsulation in commercial milk-derived EVs(S-CM EVs),but impaired EV structure,compromising transcytosis.Incubation method(I-CM EVs)preserved EVs delivery ability,but had low drug loading.Further proteomic and transmembrane studies showed that sonication greatly damaged membrane proteins involved in trans-epithelial transportation,especially endoplasmic reticulum-Golgi pathway.To overcome this dilemma,we generated a hybrid CM EVs(H-CM EVs)by fusing I-CM EVs and S-CM EVs.H-CM EVs demonstrated comparable drug encapsulation to S-CM EVs(56.14%),significantly higher than I-CM EVs(11.92%).Importantly,H-CM EVs could maintain efficient drug delivery capability by restoring membrane fluidity,repairing damaged proteins,and enhancing enzyme resistance of SCM EVs.H-CM EVs exhibited excellent absorption characteristics with 1.85-fold higher of area under the curve and 2.50-fold higher of max plasma concentration than those of SCM EVs.On typeⅠdiabetic mice,orally delivery of insulin loaded H-CM EVs and I-CM EVs showed improved hypoglycemic effects with pharmacological availabilities of 5.15%and 5.31%,which was 1.7-fold higher than that of S-CM EVs(3.00%).This H-CM EVs platform not only achieved high drug loading and maintained functionality for effective oral delivery but also highlighted the significant translational potential for improved clinical outcomes.展开更多
β-Amyloid(1-42)(Aβ_(42))can induce excessive activation of microglia,resulting in exacerbated neuroinflammation and neuronal damage.Milk fat globule-epidermal growth factor-8(MFG-E8)is known to regulate Aβ_(42)-ind...β-Amyloid(1-42)(Aβ_(42))can induce excessive activation of microglia,resulting in exacerbated neuroinflammation and neuronal damage.Milk fat globule-epidermal growth factor-8(MFG-E8)is known to regulate Aβ_(42)-induced neurotoxicity and inflammatory responses via multiple signaling pathways.However,insufficient secretion of endogenous MFG-E8 may lead to autoimmunity in the central nervous system and the accumulation of apoptotic cells.In this study,we systematically investigated the inhibitory effects and potential mechanisms of exogenously administered milk-derived MFG-E8(M-MFG-E8)on Aβ_(42)-induced inflammation in BV-2 microglia using various techniques,including cell morphology analysis,immunofluorescence staining,ELISA,q RT-PCR,and Western blot assays.The results demonstrated that Aβ_(42)significantly increased the expression levels of pro-inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in BV-2 microglia,whereas treatment with M-MFG-E8 effectively reversed these inflammatory responses.Furthermore,Aβ_(42)stimulation increased the secretion of pro-inflammatory cytokines(TNF-α,IL-1β)and the expression of the M1 marker cluster of differentiation 86(CD86),while suppressing the anti-inflammatory factors(arginase-1(Arg-1),interleukin-10(IL-10))and the M2 marker cluster of differentiation 206(CD206)in microglia.In contrast,M-MFG-E8 treatment promoted the polarization of microglia from the M1 to the M2 phenotype.Notably,M-MFG-E8 also inhibited the Aβ_(42)-induced upregulation of CD68,whereas M-MFG-E8 alone did not elicit this effect.Finally,our findings suggest that exogenous M-MFG-E8 may exert anti-inflammatory actions through the modulation of the NF-κB and PI3K/Akt pathways,providing new insights into neuronal cell repair and the development of functional foods.展开更多
The gastrointestinal tract of livestock and poultry is prone to challenge by feedborne antigens,pathogens,and other stress factors in the farm environment.Excessive physiological inflammation and oxidative stress that...The gastrointestinal tract of livestock and poultry is prone to challenge by feedborne antigens,pathogens,and other stress factors in the farm environment.Excessive physiological inflammation and oxidative stress that arises firstly disrupts the intestinal epithelial barrier followed by other components of the gastrointestinal tract.In the present review,the interrelationship between intestinal barrier inflammation and oxidative stress that contributes to the pathogenesis of inflammatory bowel disease was described.Further,the role of naturally existing immunomodulatory nutrients such as the omega-3 polyunsaturated fatty acids,citrus pectin,and milk-derived exosomes in preventing intestinal barrier inflammation was discussed.Based on the existing evidence,the possible molecular mechanism of these bioactive nutrients in the intestinal barrier was outlined for application in animal diets.展开更多
Exosomes,as promising vehicles,have been widely used in the research of oral drug delivery,but the generally low drug loading efficiency of exosomes seriously limits its application and transformation.In this study,we...Exosomes,as promising vehicles,have been widely used in the research of oral drug delivery,but the generally low drug loading efficiency of exosomes seriously limits its application and transformation.In this study,we systematically investigated the effects of drug loading methods and physicochemical properties(lipophilicity and molecular weight)on drug loading efficiency of milk-derived exosomes to explore the most appropriate loading conditions.Our finding revealed that the drug loading efficiency of exosomes was closely related to the drug loading method,drug lipophilicity,drug molecular weight and exosome/drug proportions.Of note,we demonstrated the universality that hydrophilic biomacromolecule drugs were the most appropriate loading drugs for milk-derived exosomes,which was attributed to the efficient loading capacity and sustained release behavior.Furthermore,milk-derived exosomes could significantly improve the transepithelial transport and oral bioavailability of model hydrophilic biomacromolecule drugs(octreotide,exendin-4 and salmon calcitonin).Collectively,our results suggested that the encapsulation of hydrophilic biomacromolecule drugs might be the most promising direction for milk exosomes as oral drug delivery vehicles.展开更多
Milk-derived exosomes are natural nanoparticles known for their small size,low toxicity,and high biocompat-ibility.Although several studies have examined their potential in breast cancer treatment,a comprehensive revi...Milk-derived exosomes are natural nanoparticles known for their small size,low toxicity,and high biocompat-ibility.Although several studies have examined their potential in breast cancer treatment,a comprehensive review specifically addressing the anti-breast cancer activities and underlying mechanisms of milk-derived exosomes is still lacking.This review systematically evaluates the physicochemical properties and isolation techniques of milk-derived exosomes and discusses their therapeutic efficacy and mechanistic pathways in breast cancer.Common isolation methods include differential centrifugation,size-exclusion chromatography,com-mercial exosome isolation kits,etc.The obtained milk exosomes exhibit intrinsic cytotoxicity toward cancer cells and can be co-delivered with chemopreventive compounds(e.g.,curcumin,resveratrol,cannabidiol,artemisinin,and celery extract)or chemotherapeutic agents(e.g.,doxorubicin,5-fluorouracil,paclitaxel,and oxaliplatin)to form composite nanoparticles with enhanced anti-tumor efficacy.Furthermore,they exert anti-breast cancer effects through multiple mechanisms,including induction of apoptosis,cell-cycle arrest,and modulation of NF-κB and STAT3 signaling pathways.However,further studies are needed to elucidate their detailed molecular mechanisms.Overall,this review highlights the promising potential of milk-derived exosomes as both nano-delivery platforms and standalone therapeutic agents for breast cancer treatment.展开更多
To identify milk-derived peptides with both antioxidant and calcium absorption activities in combating osteoporosis,we employed a comprehensive screening approach that included virtual enzymatic hydrolysis,molecular d...To identify milk-derived peptides with both antioxidant and calcium absorption activities in combating osteoporosis,we employed a comprehensive screening approach that included virtual enzymatic hydrolysis,molecular docking,and cellular experiments using osteoblasts.Under the optimal conditions for dual-enzyme hydrolysis,the 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)radical scavenging rate and soluble calcium binding capacity of the milk-derived peptides were 19.69% and 0.6965μg/mL,respectively.Six peptide segments,namely KEDVPSER,HKEMPFPK,YPSYG,EDVPSE,VPQLE,and IPAVF,were identified through UPLC-Q-Exactive Orbitrap MS and molecular docking for further validation.Among the peptides,YPSYG significantly promoted the proliferation of MC3T3-E1 cells both with and without CaCl_(2)(P<0.05),increasing proliferation by 38.27% and 20.67%,respectively,compared to the control group.Additionally,YPSYG significantly improved proliferation after H_(2)O_(2)-induced oxidative damage(P<0.05),with a 38.23%higher rate than the model group.Compared with rats in the osteoporosis model group,YPSYG significantly enhanced serum alkaline phosphatase(ALP)and N-terminal propeptide of type I procollagen in rats(s-PINP)levels and decreased tartrate-resistant acid phosphatase(TRAP)levels(P<0.05).Furthermore,milk-derived peptides and YPSYG significantly increased the bone weight index,maximum load,and bending energy of the femur and tibia in osteoporotic rats(P<0.05).Additionally,these peptides significantly reduced the number of osteoclasts in the metaphysis of the femur and tibia in osteoporotic rats and alleviated microstructural damage.This study confirmed that milk-derived peptides,including YPSYG,effectively promoted bone formation and improved bone microstructure in osteoporotic rats.These findings provided a foundation for developing functional foods for elderly bone health.展开更多
The imbalance between osteoblasts and osteoclasts is the cause of osteoporosis.Milk-derived extracellular vesicles(mEVs),excellent drug delivery nanocarriers,can promote bone formation and inhibit bone resorption.In t...The imbalance between osteoblasts and osteoclasts is the cause of osteoporosis.Milk-derived extracellular vesicles(mEVs),excellent drug delivery nanocarriers,can promote bone formation and inhibit bone resorption.In this study,we conjugated bone-targeting peptide(AspSerSer,DSS)6 to mEVs by click chemistry and then loaded with SRT2104,a SIRT1(silent mating-type information regulation 2 homolog 1)agonist that was proofed to help reduce bone loss.The engineered(DSS)6-mEV-SRT2104 had the intrinsic anti-osteoporosis function of mEVs and SRT2104 to reverse the imbalance in bone homeostasis by simultaneously regulating osteogenesis and osteoclastogenesis.Furthermore,we labelled mEVs with MnB nanoparticles that can be used for the in vivo magnetic resonance imaging(MRI)visualization.The obtained nanocomposites significantly prevented bone loss in osteoporosis mice and increased bone mineral density,exhibiting superior bone accumulation under MRI.We believe the proposed(DSS)6-mEV-SRT2104/MnB provides a novel paradigm for osteoporosis treatment and monitoring.展开更多
As endogenous courier vesicles,exosomes play crucial roles in macromolecule transmission and intercellular communication.Therefore,exosomes have drawn increasing attention as biomimetic drug-delivery vehicles over the...As endogenous courier vesicles,exosomes play crucial roles in macromolecule transmission and intercellular communication.Therefore,exosomes have drawn increasing attention as biomimetic drug-delivery vehicles over the past few years.However,few studies have investigated the encapsulation of peptide/protein drugs into exosomes for oral administration.Additionally,the mechanisms underlying their biomimetic properties as oral delivery vehicles remain unknown.Herein,insulin-loaded milk-derived exosomes(EXO@INS)were fabricated and the in vivo hypoglycemic effect was investigated on type I diabetic rats.Surprisingly,EXO@INS(50 and 30 IU/kg)elicited a more superior and more sustained hypoglycemic effect compared with that obtained with subcutaneously injected insulin.Further mechanism studies indicated that the origin of excellent oral-performance of milk-derived exosomes combined active multi-targeting uptake,pH adaptation during gastrointestinal transit,nutrient assimilation related ERK1/2 and p38 MAPK signal pathway activation and intestinal mucus penetration.This study provides the first demonstration that multifunctional milk-derived exosomes offer solutions to many of the challenges arising from oral drug delivery and thus provide new insights into developing naturally-equipped nanovehicles for oral drug administration.展开更多
Oral administration facilitates the direct delivery of drugs to lesions within the small intestine and colon,making it an ideal approach for treating patients with inflammatory bowel disease.However,multiple physical ...Oral administration facilitates the direct delivery of drugs to lesions within the small intestine and colon,making it an ideal approach for treating patients with inflammatory bowel disease.However,multiple physical barriers impede the delivery of oral RNA drugs through the gastrointestinal tract.Herein,we developed a novel oral siRNA delivery system that protects nucleic acids in extreme environments by employing exosomes derived from milk to encapsulate tumor necrosis factor-alpha(TNF-α)siRNA completely.The remarkable structural stability of milk-derived exosomes(M-Exos),as opposed to those from HEK293T cells,makes them exceptional siRNA carriers.Results demonstrate that milk exosomes loaded with TNF-αsiRNA(M-Exo/siR)can effectively inhibit the expression of TNF-α-related inflammatory cytokines.Moreover,given that milk exosomes are composed of unique lipids with high bioavailability,orally administered M-Exo/siR effectively reach colonic tissues,leading to decreased TNF-αexpression and successful alleviation of colitis symptoms in a dextran sulfate sodium-induced inflammatory bowel disease murine model.Hence,milk-derived exosomes carrying TNF-αsiRNA can be effectively employed to treat inflammatory bowel disease.Indeed,using exosomes naturally derived from milk may shift the current paradigm of oral gene delivery,including siRNA.展开更多
Isolation of high-purity milk-derived extracellular vesicles(miEV)is complicated by protein impurities,such as casein.The focus of this study was to investigate whether the treatment of milk with acetic acid(AA),ethyl...Isolation of high-purity milk-derived extracellular vesicles(miEV)is complicated by protein impurities,such as casein.The focus of this study was to investigate whether the treatment of milk with acetic acid(AA),ethylenediaminetetraacetic acid(EDTA),and sodium citrate(SC),which are known chemicals to deplete caseins from miEV,alters their physicochemical and functional characteristics differently.For this purpose,following the treatment of milk with AA,EDTA,and SC,various subpopulations(12 K,35 K,70 K,and 100 K)of miEV were isolated by differential ultracentrifugation and characterized by protein quantification assay,gel electrophoresis,western blotting,dynamic light scattering,and scanning electron microscopy.In addition,the cellular uptake and immunomodulatory potential of miEV were also assessed.The results indicated that protein impurities,including caseins,were differentially depleted from miEV subpopulations by AA,EDTA,and SC.However,the acidification of milk induced partial protein aggregation.Additionally,miEV from both AA-and EDTA-treated milk exhibited a less smooth surface and higher uptake compared to those from SC-treated milk.Although all miEV significantly reduced T cell proliferation,a higher immunomodulatory potential was found in miEV from AA-treated milk.In conclusion,AA,SC,and EDTA have different efficiencies in improving the purity of miEV and can affect their physicochemical and functional characteristics differently.This issue may be critical for the efficient isolation of pure subpopulations of miEV for biomedical research.展开更多
基金supported by the National Natural Science Foundation of China(no.32372838,U22A20506)the National Key Research and Development Program of China(no.2024YFD1300104)+1 种基金the scientific and technological development program of Jilin province(YDZJ202203CGZH037)the earmarked fund for JLARS-2025-070203。
文摘Background Exosomes are crucial mediators of intercellular communication.As a key component of milk,milkderived exosomes are abundant in genetic cargo,particularly micro RNAs(mi RNAs),indicating their potential role in regulating mammary gland physiology.Therefore,this study aimed to investigate the specificity of mi RNAs in milkderived exosomes and their regulatory roles in lipid synthesis in bovine mammary epithelial cells(BMECs).Results Based on 17,838 DHI records showing a significantly higher milk fat percentage(MFP)in late lactation(4.24%±1.07%),10 high-(5.96%±0.26%,HMF)and 10 low-MFP(1.68%±0.23%,LMF)cows were selected during this stage for milk-derived exosome isolation and mi RNA profiling.Exosomes isolated via differential ultracentrifugation were verified as 50-150 nm vesicles expressing CD9,CD81,and TSG101.mi RNA sequencing identified 1,320 differentially expressed mi RNAs(496 upregulated and 824 downregulated)between the HMF_EXO and LMF_EXO groups.Uptake assays confirmed that BMECs internalized these exosomes,and q RT-PCR validation showed that mi R-423-5p and mi R-125b were significantly upregulated and downregulated in HMF_EXO-and LMF_EXO-treated BMECs,respectively.Functionally,exosomal mi R-423-5p promoted intracellular lipid accumulation and TG synthesis in BMECs by targeting APOA5,whereas mi R-125b inhibited lipolysis and fatty acid oxidation by repressing SLC27A1.Conclusions This study demonstrates that milk-derived exosomal mi RNAs represent a novel mechanism for regulating milk fat synthesis.Specifically,mi R-423-5p and mi R-125b directly modulated lipid metabolism in BMECs via the mi R-423-5p/APOA5 and mi R-125b/SLC27A1 pathways.These findings provide new insights into the molecular regulation of milk fat synthesis and highlight the importance of exosome-mediated intercellular communication in the lactating mammary gland.
基金financial support from the the Regional Innovation and Development Joint Fund of National Natural Science Foundation of China(grant numbers:U22A20356)the National Key R&D Program of China(No.2021YFE0115200)the National Natural Science Foundation of China(No.81872818).
文摘Milk-derived extracellular vesicles(EVs)are promising for oral drug delivery,yet different loading methods exhibit distinct impacts on drug encapsulation and membrane integrity.This study demonstrated that sonication method achieved high drug encapsulation in commercial milk-derived EVs(S-CM EVs),but impaired EV structure,compromising transcytosis.Incubation method(I-CM EVs)preserved EVs delivery ability,but had low drug loading.Further proteomic and transmembrane studies showed that sonication greatly damaged membrane proteins involved in trans-epithelial transportation,especially endoplasmic reticulum-Golgi pathway.To overcome this dilemma,we generated a hybrid CM EVs(H-CM EVs)by fusing I-CM EVs and S-CM EVs.H-CM EVs demonstrated comparable drug encapsulation to S-CM EVs(56.14%),significantly higher than I-CM EVs(11.92%).Importantly,H-CM EVs could maintain efficient drug delivery capability by restoring membrane fluidity,repairing damaged proteins,and enhancing enzyme resistance of SCM EVs.H-CM EVs exhibited excellent absorption characteristics with 1.85-fold higher of area under the curve and 2.50-fold higher of max plasma concentration than those of SCM EVs.On typeⅠdiabetic mice,orally delivery of insulin loaded H-CM EVs and I-CM EVs showed improved hypoglycemic effects with pharmacological availabilities of 5.15%and 5.31%,which was 1.7-fold higher than that of S-CM EVs(3.00%).This H-CM EVs platform not only achieved high drug loading and maintained functionality for effective oral delivery but also highlighted the significant translational potential for improved clinical outcomes.
基金the financial support received from the Science and Technology Planning Project of Sichuan Province(Jiebangguashuai Project)(2023YFN0101)。
文摘β-Amyloid(1-42)(Aβ_(42))can induce excessive activation of microglia,resulting in exacerbated neuroinflammation and neuronal damage.Milk fat globule-epidermal growth factor-8(MFG-E8)is known to regulate Aβ_(42)-induced neurotoxicity and inflammatory responses via multiple signaling pathways.However,insufficient secretion of endogenous MFG-E8 may lead to autoimmunity in the central nervous system and the accumulation of apoptotic cells.In this study,we systematically investigated the inhibitory effects and potential mechanisms of exogenously administered milk-derived MFG-E8(M-MFG-E8)on Aβ_(42)-induced inflammation in BV-2 microglia using various techniques,including cell morphology analysis,immunofluorescence staining,ELISA,q RT-PCR,and Western blot assays.The results demonstrated that Aβ_(42)significantly increased the expression levels of pro-inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in BV-2 microglia,whereas treatment with M-MFG-E8 effectively reversed these inflammatory responses.Furthermore,Aβ_(42)stimulation increased the secretion of pro-inflammatory cytokines(TNF-α,IL-1β)and the expression of the M1 marker cluster of differentiation 86(CD86),while suppressing the anti-inflammatory factors(arginase-1(Arg-1),interleukin-10(IL-10))and the M2 marker cluster of differentiation 206(CD206)in microglia.In contrast,M-MFG-E8 treatment promoted the polarization of microglia from the M1 to the M2 phenotype.Notably,M-MFG-E8 also inhibited the Aβ_(42)-induced upregulation of CD68,whereas M-MFG-E8 alone did not elicit this effect.Finally,our findings suggest that exogenous M-MFG-E8 may exert anti-inflammatory actions through the modulation of the NF-κB and PI3K/Akt pathways,providing new insights into neuronal cell repair and the development of functional foods.
基金a European Joint Doctorate Degree programme in Molecular Animal Nutrition(MANNA)between University of Milan(Italy)and University of Veterinary Medicine and Pharmacy in Kosice(Slovakia)funding from the European Union’s Horizon 2020 programme under the Marie Slodowska-Curie Grant agreement No 765423.
文摘The gastrointestinal tract of livestock and poultry is prone to challenge by feedborne antigens,pathogens,and other stress factors in the farm environment.Excessive physiological inflammation and oxidative stress that arises firstly disrupts the intestinal epithelial barrier followed by other components of the gastrointestinal tract.In the present review,the interrelationship between intestinal barrier inflammation and oxidative stress that contributes to the pathogenesis of inflammatory bowel disease was described.Further,the role of naturally existing immunomodulatory nutrients such as the omega-3 polyunsaturated fatty acids,citrus pectin,and milk-derived exosomes in preventing intestinal barrier inflammation was discussed.Based on the existing evidence,the possible molecular mechanism of these bioactive nutrients in the intestinal barrier was outlined for application in animal diets.
基金The authors gratefully acknowledge financial support from National Natural Science Foundation of China(81872818)National Key R&D Program of China(2021YFE0115200).
文摘Exosomes,as promising vehicles,have been widely used in the research of oral drug delivery,but the generally low drug loading efficiency of exosomes seriously limits its application and transformation.In this study,we systematically investigated the effects of drug loading methods and physicochemical properties(lipophilicity and molecular weight)on drug loading efficiency of milk-derived exosomes to explore the most appropriate loading conditions.Our finding revealed that the drug loading efficiency of exosomes was closely related to the drug loading method,drug lipophilicity,drug molecular weight and exosome/drug proportions.Of note,we demonstrated the universality that hydrophilic biomacromolecule drugs were the most appropriate loading drugs for milk-derived exosomes,which was attributed to the efficient loading capacity and sustained release behavior.Furthermore,milk-derived exosomes could significantly improve the transepithelial transport and oral bioavailability of model hydrophilic biomacromolecule drugs(octreotide,exendin-4 and salmon calcitonin).Collectively,our results suggested that the encapsulation of hydrophilic biomacromolecule drugs might be the most promising direction for milk exosomes as oral drug delivery vehicles.
基金supported by National Natural Science Foundation of China(No.82172356,No.82203559)Natural Science Foundation of Shenzhen,China(JCYJ20230807115112024)Guangdong Basic and Applied Basic Research Foundation(2023A1515220238).
文摘Milk-derived exosomes are natural nanoparticles known for their small size,low toxicity,and high biocompat-ibility.Although several studies have examined their potential in breast cancer treatment,a comprehensive review specifically addressing the anti-breast cancer activities and underlying mechanisms of milk-derived exosomes is still lacking.This review systematically evaluates the physicochemical properties and isolation techniques of milk-derived exosomes and discusses their therapeutic efficacy and mechanistic pathways in breast cancer.Common isolation methods include differential centrifugation,size-exclusion chromatography,com-mercial exosome isolation kits,etc.The obtained milk exosomes exhibit intrinsic cytotoxicity toward cancer cells and can be co-delivered with chemopreventive compounds(e.g.,curcumin,resveratrol,cannabidiol,artemisinin,and celery extract)or chemotherapeutic agents(e.g.,doxorubicin,5-fluorouracil,paclitaxel,and oxaliplatin)to form composite nanoparticles with enhanced anti-tumor efficacy.Furthermore,they exert anti-breast cancer effects through multiple mechanisms,including induction of apoptosis,cell-cycle arrest,and modulation of NF-κB and STAT3 signaling pathways.However,further studies are needed to elucidate their detailed molecular mechanisms.Overall,this review highlights the promising potential of milk-derived exosomes as both nano-delivery platforms and standalone therapeutic agents for breast cancer treatment.
基金financially supported by Chinese Nutrition Society(CNS)Nutrition Science Foundation-Feihe Tizhi,nutrition and health research fund(CNS-Feihe2021-38).
文摘To identify milk-derived peptides with both antioxidant and calcium absorption activities in combating osteoporosis,we employed a comprehensive screening approach that included virtual enzymatic hydrolysis,molecular docking,and cellular experiments using osteoblasts.Under the optimal conditions for dual-enzyme hydrolysis,the 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)radical scavenging rate and soluble calcium binding capacity of the milk-derived peptides were 19.69% and 0.6965μg/mL,respectively.Six peptide segments,namely KEDVPSER,HKEMPFPK,YPSYG,EDVPSE,VPQLE,and IPAVF,were identified through UPLC-Q-Exactive Orbitrap MS and molecular docking for further validation.Among the peptides,YPSYG significantly promoted the proliferation of MC3T3-E1 cells both with and without CaCl_(2)(P<0.05),increasing proliferation by 38.27% and 20.67%,respectively,compared to the control group.Additionally,YPSYG significantly improved proliferation after H_(2)O_(2)-induced oxidative damage(P<0.05),with a 38.23%higher rate than the model group.Compared with rats in the osteoporosis model group,YPSYG significantly enhanced serum alkaline phosphatase(ALP)and N-terminal propeptide of type I procollagen in rats(s-PINP)levels and decreased tartrate-resistant acid phosphatase(TRAP)levels(P<0.05).Furthermore,milk-derived peptides and YPSYG significantly increased the bone weight index,maximum load,and bending energy of the femur and tibia in osteoporotic rats(P<0.05).Additionally,these peptides significantly reduced the number of osteoclasts in the metaphysis of the femur and tibia in osteoporotic rats and alleviated microstructural damage.This study confirmed that milk-derived peptides,including YPSYG,effectively promoted bone formation and improved bone microstructure in osteoporotic rats.These findings provided a foundation for developing functional foods for elderly bone health.
基金supported by National Natural Science Foundation of China(52103198,52311530079)Basic and Applied Basic Research Foundation of Guangdong Province(2023A1515010067)Natural Science Foundation of Shenzhen Municipality(RCBS20210609104333005,JCYJ20220530145001003).
文摘The imbalance between osteoblasts and osteoclasts is the cause of osteoporosis.Milk-derived extracellular vesicles(mEVs),excellent drug delivery nanocarriers,can promote bone formation and inhibit bone resorption.In this study,we conjugated bone-targeting peptide(AspSerSer,DSS)6 to mEVs by click chemistry and then loaded with SRT2104,a SIRT1(silent mating-type information regulation 2 homolog 1)agonist that was proofed to help reduce bone loss.The engineered(DSS)6-mEV-SRT2104 had the intrinsic anti-osteoporosis function of mEVs and SRT2104 to reverse the imbalance in bone homeostasis by simultaneously regulating osteogenesis and osteoclastogenesis.Furthermore,we labelled mEVs with MnB nanoparticles that can be used for the in vivo magnetic resonance imaging(MRI)visualization.The obtained nanocomposites significantly prevented bone loss in osteoporosis mice and increased bone mineral density,exhibiting superior bone accumulation under MRI.We believe the proposed(DSS)6-mEV-SRT2104/MnB provides a novel paradigm for osteoporosis treatment and monitoring.
基金We gratefully acknowledge financial support from the National Science Foundation for Distinguished Young Scholars(81625023,China)the National Natural Science Foundation of China(81872818)the Major Research Plan of National Natural Science Foundation of China(81690261).
文摘As endogenous courier vesicles,exosomes play crucial roles in macromolecule transmission and intercellular communication.Therefore,exosomes have drawn increasing attention as biomimetic drug-delivery vehicles over the past few years.However,few studies have investigated the encapsulation of peptide/protein drugs into exosomes for oral administration.Additionally,the mechanisms underlying their biomimetic properties as oral delivery vehicles remain unknown.Herein,insulin-loaded milk-derived exosomes(EXO@INS)were fabricated and the in vivo hypoglycemic effect was investigated on type I diabetic rats.Surprisingly,EXO@INS(50 and 30 IU/kg)elicited a more superior and more sustained hypoglycemic effect compared with that obtained with subcutaneously injected insulin.Further mechanism studies indicated that the origin of excellent oral-performance of milk-derived exosomes combined active multi-targeting uptake,pH adaptation during gastrointestinal transit,nutrient assimilation related ERK1/2 and p38 MAPK signal pathway activation and intestinal mucus penetration.This study provides the first demonstration that multifunctional milk-derived exosomes offer solutions to many of the challenges arising from oral drug delivery and thus provide new insights into developing naturally-equipped nanovehicles for oral drug administration.
基金supported by the Bio&Medical Technology Development Program(NRF-2022M3E5F2018170)the Intramural Research Program of the Korea Institute of Science and Technology(KIST).
文摘Oral administration facilitates the direct delivery of drugs to lesions within the small intestine and colon,making it an ideal approach for treating patients with inflammatory bowel disease.However,multiple physical barriers impede the delivery of oral RNA drugs through the gastrointestinal tract.Herein,we developed a novel oral siRNA delivery system that protects nucleic acids in extreme environments by employing exosomes derived from milk to encapsulate tumor necrosis factor-alpha(TNF-α)siRNA completely.The remarkable structural stability of milk-derived exosomes(M-Exos),as opposed to those from HEK293T cells,makes them exceptional siRNA carriers.Results demonstrate that milk exosomes loaded with TNF-αsiRNA(M-Exo/siR)can effectively inhibit the expression of TNF-α-related inflammatory cytokines.Moreover,given that milk exosomes are composed of unique lipids with high bioavailability,orally administered M-Exo/siR effectively reach colonic tissues,leading to decreased TNF-αexpression and successful alleviation of colitis symptoms in a dextran sulfate sodium-induced inflammatory bowel disease murine model.Hence,milk-derived exosomes carrying TNF-αsiRNA can be effectively employed to treat inflammatory bowel disease.Indeed,using exosomes naturally derived from milk may shift the current paradigm of oral gene delivery,including siRNA.
基金supported by Tehran University of Medical Science(Grant No.1400-03-148-55020)Royan Institute(401000177).
文摘Isolation of high-purity milk-derived extracellular vesicles(miEV)is complicated by protein impurities,such as casein.The focus of this study was to investigate whether the treatment of milk with acetic acid(AA),ethylenediaminetetraacetic acid(EDTA),and sodium citrate(SC),which are known chemicals to deplete caseins from miEV,alters their physicochemical and functional characteristics differently.For this purpose,following the treatment of milk with AA,EDTA,and SC,various subpopulations(12 K,35 K,70 K,and 100 K)of miEV were isolated by differential ultracentrifugation and characterized by protein quantification assay,gel electrophoresis,western blotting,dynamic light scattering,and scanning electron microscopy.In addition,the cellular uptake and immunomodulatory potential of miEV were also assessed.The results indicated that protein impurities,including caseins,were differentially depleted from miEV subpopulations by AA,EDTA,and SC.However,the acidification of milk induced partial protein aggregation.Additionally,miEV from both AA-and EDTA-treated milk exhibited a less smooth surface and higher uptake compared to those from SC-treated milk.Although all miEV significantly reduced T cell proliferation,a higher immunomodulatory potential was found in miEV from AA-treated milk.In conclusion,AA,SC,and EDTA have different efficiencies in improving the purity of miEV and can affect their physicochemical and functional characteristics differently.This issue may be critical for the efficient isolation of pure subpopulations of miEV for biomedical research.
