Effect of Ranae Oviductus Protein Hydrolysate on Ethanol-induced L-02 Cell Injury / 中国实验方剂学杂志

Objective:To study the protective effect and mechanism of Ranae Oviductus protein hydrolysate （ROPH） on the expression of pathway-related proteins in ethanol-induced L-02 cell injury. Method:The ROPH was prepared by compound enzymatic hydrolysis. L-02 cell injury model was induced with 400 mmol·L^-1ethanol. Cell viability was detected by cell counting kit-8 （CCK-8） assay. Cell cycle and apoptosis were examined by flow cytometry. JC-1/Hochest staining was employed for qualitative investigation. The expression of related proteins in apoptosis， mitogen-activated protein kinase （MAPK） signaling pathway， and pyroptosis in L-02 cells was detected by Western blot. Result:The results of the CCK-8 assay showed that 400 mmol·L^-1ethanol could induce L-02 cell injury within 12 hours. Compared with the blank group， the model group showed decreased viability of L-02 cells （<italic>P</italic><0.01）， elevated percentage of the cell cycle in the G₀/G₁ phase （<italic>P</italic><0.01）， increased total cell apoptosis rate （<italic>P</italic><0.01）， reduced mitochondrial membrane potential （<italic>P</italic><0.01）， up-regulated expression of apoptosis-related proteins ［B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Cytochrome C （Cyt C）， and cysteine-dependent aspartate specific protease-3 （Caspase-3）］ （<italic>P</italic><0.05， <italic>P</italic><0.01） and MAPK signaling pathway-related proteins ［C-Jun amino-terminal kinase （JNK） and p38 MAPK］ （<italic>P</italic><0.05， <italic>P</italic><0.01）， and potentiated expression of pyrolysis-related proteins Caspase-1 and interleukin-1<italic>β </italic>（IL-1<italic>β</italic>） （<italic>P</italic><0.05）. Compared with the model group， the ROPH treatment group exhibited improved cell cycle arrest （<italic>P</italic><0.05， <italic>P</italic><0.01）， diminished total cell apoptosis rate （<italic>P</italic><0.01）， elevated mitochondrial membrane potential in a dose-dependent manner， down-regulated expression of Bax， Cyt C， and Caspase-3 proteins （<italic>P</italic><0.05， <italic>P</italic><0.01）， up-regulated expression of Bcl-2 protein （<italic>P</italic><0.05， <italic>P</italic><0.01）， and a downward trend in expression of proteins related to MAPK signaling pathway and pyrolysis （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:ROPH could inhibit oxidative stress-triggered liver injury in ethanol-induced cells by improving mitochondrial membrane potential， reducing the expression of proteins in the mitochondria-mediated apoptosis pathway， and inhibiting the expression of proteins related to the MAPK signaling pathway and pyrolysis pathway to reduce the mitochondrial dysfunction and inflammatory response in ethanol-induced L-02 liver cells and inhibit oxidative stress， thereby exerting a therapeutic role in alcoholic liver injury.

Two novel antimicrobial and anticancer peptides prediction from Vibrio sp. strain ES25

Proteins from microbes are a large source of potential raw material for novel pharmaceuticals. Endophytic bacteriaassociated with algae associated have been shown to be particularly promising candidates. In a previous study, a crudeprotein extract was isolated from an endophytic Vibrio sp. strain ES25. The extract was found to have anti-microbialactivity. Despite these promising results, further work was needed to characterize both the protein and the bacteria thatproduce it. In this study, we sought to increase the activity of the previously discovered protein through pepsin digestionas well as provide sequence data to characterize the bacterial isolate. Here, we show promising evidence that thehydrolysate produced (QA1d) was found to have a promising antimicrobial as well as anticancer properties. Through theDNA sequencing, we also provide better taxonomic information on the isolate. Finally using Liquid ChromatographyMass Spectroscopy/Mass Spectroscopy (LC-MS/MS) and in silico prediction, we identify to novel peptides producedfrom the bacterial protein that were predicted to have multifunctional activity. These results shown here indicate thesepeptides are promising candidates for anticancer drug development or antibacterial agents for the food industry.

Antioxidant and cytotoxic properties of protein hydrolysates obtained from enzymatic hydrolysis of Klunzinger's mullet (Liza klunzingeri) muscle

Today, consumers are looking for functional foods that promote health and prevent certain diseases in addition to provide nutritional requirements. This study aimed to evaluate the antioxidant and cytotoxic properties of Liza klunzingeri protein hydrolysates. Fish protein hydrolysates (FPHs) were prepared from L. klunzingeri muscle using enzymatic hydrolysis with papain at enzyme/substrate ratios of 1:25 and 1:50 for 45, 90 and 180 min. The antioxidant activities of the FPHs were investigated through five antioxidant assays. The cytotoxic effects on 4T1 carcinoma cell line were also evaluated. The amino acid composition and molecular weight distribution of the hydrolysate with the highest antioxidant activity were determined by HPLC. All six FPHs exhibited good scavenging activity on ABTS (IC50=0.60-0.12 mg/mL), DPPH (IC50= 3.18-2.08 mg/mL), and hydroxyl (IC50=4.13-2.07 mg/mL) radicals. They also showed moderate Fe+2 chelating capacity (IC50=2.12-12.60 mg/mL) and relatively poor ferric reducing activity (absorbance at 70 nm= 0.01-0.15, 5 mg/mL). In addition, all hydrolysates showed cytotoxic activities against the 4T1 cells (IC50=1.62-2.61 mg/mL). 94.6% of peptide in hydrolysate with the highest antioxidant activity had molecular weight less than 1,000 Da. L. klunzingeri protein hydrolysates show significant antioxidant and anticancer activities in vitro and are suggested to be used in animal studies.

Analysis of the Efficacy of Brain Protein Hydrolysate Injection in the Treatment of Elderly Patients with Parkinson / 现代生物医学进展

Objective:To investigate the efficacy and safety of brain protein hydrolysate injection in the treatment of elderly patients with parkinson.Methods:120 patients with parkinson were selected and randomly divided into two groups.The control group (57 cases) was treated by routine treatment,while the observation group (63 cases) was given brain protein hydrolysate injection on the basis of routine treatment.The UPDRS scores,SF-36 scores and adverse reactions during treatment were observed and recorded.Results:Before treatment,the UPDRS and SF-36 scores of both groups had no significant difference (P＞0.05).After treatment,the UPDRS scores of both groups were significantly decreased,and the UPDRS score of observation group was lower than that of the control group(P＜0.05).The social function,emotional function,mental health score of observation group were higher than those of the control group(P＜0.05).During treatment,1 cases of dizziness,1 cases of nausea were found in the control group.The adverse reaction rate was 3.5％.2 cases of insomnia,1 cases of fatigue,1 cases of dizziness and 1 cases of nausea were found in the observation group.The adverse reaction rate was 7.9％.There was no statistical difference in the incidence of adverse reactions between two groups (P＞0.05).Conclusion:Brain protein hydrolysate injection had significant effect on the Parkinson.It could improve the patients' thinking ability,mental health and quality of life with high safety.

Therapeutic effect of Xiefu Zhuyu decoction joint brain protein hydrolysate injection in traumatic brain injury and its effect on cerebrospinal fluid of ET-1 / 中国生化药物杂志

Objective To explore the therapeutic effect of Xiefu Zhuyu decoction joint brain protein hydrolysate injection in the treatment of patients with traumatic brain injury and its effect on cerebrospinal fluid of endothelin-1 ( ET-1 ) .Methods 102 cases of patients with craniocerebral injury were divided into control group (n=51) and experimental group (n=51),the control group were treated with brain protein hydrolyzate injection, the experimental group based on the control group were treated with Xuefu Zhuyu decoction, the curative effect, ET-1, inflammatory factor, protein, blood rheology, S-100β, neuron specific enolase (NSE), National Institutes of Health Stroke Scale (NIHSS) and complications of two groups were compared.Results The total effective rate of experimental group was higher than the control group (94.11% vs.78.43%), the difference was statistically significant (P<0.05).The ET-1 of experimental group was lower than the control group[(32.45 ±4.05) ng/L vs.(28.11 ±3.50) ng/L] (P<0.05).The inflammatory factor, protein, blood rheology, NSE, S-100βof experimental group were better than those of control group (P<0.05). The NIHSS of experimental group was lower than that of the control group ( P <0.05 ) .The complication rate had no difference between two groups. Conclusion The clinical curative effect of Xiefu Zhuyu decoction joint brain protein hydrolysate injection in the treatment of traumatic brain injury , can reduce cerebrospinal fluid levels of ET-1 and can improve inflammatory factors, proteins and hemorheology.

Antioxidant and Anti-aging Activities of Silybum Marianum Protein Hydrolysate in Mice Treated with D-galactose / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>In the present study, we investigated the antioxidant and anti-aging effects of Silybum marianum protein hydrolysate (SMPH) in D-galactose-treated mice.</p><p><b>METHODS</b>D-galactose (500 mg/kg body weight) was intraperitoneally injected daily for 7 weeks to accelerate aging, and SMPH (400, 800, 1,200 mg/kg body weight, respectively) was simultaneously administered orally. The antioxidant and anti-aging effects of SMPH in the liver and brain were measured by biochemical assays. Transmission electron microscopy (TEM) was performed to study the ultrastructure of liver mitochondri.</p><p><b>RESULTS</b>SMPH decreased triglyceride and cholesterol levels in the D-galactose-treated mice. It significantly elevated the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC), which were suppressed by D-galactose. Monoamine oxidase (MAO) and malondialdehyde (MDA) levels as well as the concentrations of caspase-3 and 8-OHdG in the liver and brain were significantly reduced by SMPH. Moreover, it increased Bcl-2 levels in the liver and brain. Furthermore, SMPH significantly attenuated D-galactose-induced liver mitochondrial dysfunction by improving the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase as well as mitochondrial membrane potential (ΔΨm) and fluidity. TEM showed that the degree of liver mitochondrial damage was significantly decreased by SMPH.</p><p><b>CONCLUSION</b>The results indicated that SMPH protects against D-galactose-induced accelerated aging in mice through its antioxidant and anti-aging activities.</p>

Antimicrobial Activity of Protein Hydrolysate from Marine Molluscs Babylonia spirata (Linnaeus, 1758).

The present study is to investigate the antimicrobial activity of protein hydrolysate of marine water mollusks Babylonia spirata (Linnaeus, 1758). Protein hydrolysate was prepared from tissue of Babylonia spirata by enzymatic hydrolysis. Enzyme digestion were carried out with the enzyme Trypsin. The protein concentration was estimated by Bradford’s method and the protein quantification was done by using SDS PAGE analysis. Antibacterial assay was carried out against four bacterial pathogens by agar well diffusion method and antifungal activity was performed against three human pathogenic fungal strains. 2.6mg/ml protein concentration was estimated by Bradford’s method and 40 to 200 kDa protein bands were resulted in SDS PAGE analysis. In antimicrobial activity, the maximum zone of inhibition was observed against Staphylococcus aureus22.16 +1.04mm at 1000μg/ml concentration and the maximum zone of inhibition was observed in Aspergillus fumigatus13.5+0.5 in 1000μg/ml concentration. These results are signify that the protein hydrolysate of marine molluscs Babylonia spirata express remarkable antimicrobial activity.

Hidrolisado proteico da semente de cupuaçu como fonte de peptídeos inibidores da enzima conversora da angiotensina I / Hydrolyzed cupuassu seed protein as a source of angiotensin I- converting enzyme inhibitory peptides

Peptídeos com ação inibitória da enzima conversora da angiotensina I (ECA) podem ser obtidos a partir de diversos alimentos e exercer efeito anti-hipertensivo. O cupuaçu (Theobroma grandiflorum S.), fruto nativo da Amazônia, possui sementes comestíveis contendo proteína de reserva similar à do cacau (Theobroma cacao L.), as quais parecem ser fonte de peptídeos inibidores da ECA. Desse modo, o objetivo deste trabalho foi investigar in vitro a ocorrência de peptídeos inibidores da ECA no hidrolisado proteico da semente de cupuaçu obtido por ação da Alcalase. O hidrolisado revelou o desaparecimento de proteínas entre 27 a 180 kDa, incluindo as globulinas, e o surgimento daquelas abaixo de 15 kDa após 2 h de hidrólise, indicando a formação de peptídeos menores. O ensaio de atividade utilizando o substrato Abz-FRK(Dnp)-P-OH revelou que o hidrolisado total promoveu 65% de inibição da ECA e esse pool peptídico foi fracionado em cinco frações (F1-F5) por cromatografia em fase reversa (RP-HPLC). Após a etapa de purificação, o monitoramento da inibição apontou, ao final, duas frações (3.2.8 e 3.4.10) com maior atividade inibitória. Oito peptídeos foram identificados por LC-MS/MS, sendo três deles já conhecidos como inibidores da ECA. Outros cinco novos peptídeos identificados (FLEK, GSGKHVSP, LDNK, MVVDKLF e MEKHS) foram sintetizados e tiveram sua ação inibitória validada por ensaios in vitro. O peptídeo GSGKHVSP apresentou a menor IC50 (3,11 µM) e Ki (0,74 µM), sendo um inibidor tipo misto quanto ao seu mecanismo de inibição revelado pelo gráfico de Lineweaver-Burk. Os resultados permitem concluir que o isolado proteico da semente de cupuaçu pode ser uma fonte para obtenção de peptídeos anti-hipertensivos, a despeito de serem necessárias investigações sobre a resistência desses peptídeos à digestão gastrointestinal e a eficácia da inibição in vivo

Peptides with angiotensin I-converting enzyme (ACE) inhibitory activity may be obtained from several foods and cause antihypertensive effect. Cupuassu (Theobroma grandiflorum S.), a native fruit from Amazon, has edible seeds with a storage protein similar to that of cocoa (Theobroma cacao L.) which seems to have incrypted ACE inhibitor peptides. Thus, the aim of this project was to investigate the in vitro formation of ACE inhibitory peptides in protein hydrolysate from cupuassu seeds using Alcalase enzyme. The hydrolysate revealed the disappearance of proteins between 27 and 181 kDa after 2h hydrolysis, including the globulin, and the increase of those below 15 kDa, indicating the formation of peptides. The ACE inhibitory activity assays using the substrate Abz-FRK(Dnp)P-OH revealed the hydrolysate had 65% ACE inhibition and the pool of peptides was fractionated into five fractions (F1-F5) by reversed phase high-performance liquid chromatography (RP-HPLC). After the purification step, two fractions (3.2.8 e 3.4.10) exhibited the highest ACE-inhibitory activity. Eight peptides had been identified by LC-MS/MS and three of them were ACE inhibitors. The other newly identified peptides (FLEK, GSGKHVSP, LDNK, MVVDKLF and MEKHS) were synthesized and in vitro assayed for ACE inhibitory activity. The peptide GSGKHVSP had the lower IC50 (3.11 µM) and Ki (0.74 µM). Lineweaver-Burk plots suggest this peptide is a mixed-type inhibitor according to the inhibition mechanism. The results indicate that protein isolate from cupuassu seeds may be a good protein source of antihypertensive peptides and further investigation is needed in order to evaluate the resistance of these peptides to gastrointestinal digestion and the inhibitory activity in vivo

Process study of brain protein hydrolysate of inactivate and virus removal / 中国基层医药

Objective To study the process of brain protein hydrolysate of inactivate and virus removal.Methods The Parvoviridae parvovirus genera of porcine parvovirus (PPV),vesicular stomatitis virus rhabdovirus genera of vesicular stomatitis virus (VSV) were chosen as a model virus,wherein PPV represents no envelope deoxyribonucleic acid(DNA) virus,VSV represents the envelope ribonucleic acid(RNA) virus.Simulation of the production process of virus inactivation steps 100 ℃ × 30 min,ultrafiltration as inactivation/removal condition.The virus respectively according to 1 ∶ 9 into the brain protein hydrolysate,high temperature and ultra filtration virus inactivation/removal.In pig kidney cells (PK-15) in PPV cell culture,Africa green monkey kidney cells(Vero cells) cultured VSV,determination of virus titer.Results PPV and VSV through the sterilization,virus median tissue culture infective dose(TCID50) were 6.15log/0.1mL(logs),5.37 log/0.1mL(logs) ;removal processaverage virus reduction coefficient were 6.15log/0.1mL(logs),5.37 log/0.1mL(logs).Conclusion The high temperature and ultra filtration produces brain protein hydrolysate solution process are effective virus inactivation/removal process.

Protein hydrolysate formula in newborns / 国际儿科学杂志

Enteral feeding is essential to the newborns,especially premature newborns,which related to infants′survival and long-term prognosis. There are different kinds of infant formulas which can be chosed in case of maternal breast milk is not available,but how to choose has always been the focus of attention. In recent years protein hydrolysate formula has been applied expanded gradually in the clinical practice,and more related research has been completed,which have conducted yielding new insights for researchers. Some recent applica-tion of protein hydrolysate formula in the neonatal are reviewed and discussed.

EFFECT OF CHICKPEA PROTEIN HYDROLYSATE ON ANTIOXIDANT ABILITIES IN AGED MICE INDUCED BY D-GALATOSE / 营养学报

Objective To study the effects of chickpea protein hydrolysate(CPH) in different doses on of antioxidant abilities in serum,liver and heart in aged mice induced by D-galactose .Method The subacute aged model mice were made by neck back subcutaneous injection of D-galactose continually.The content of malondialdehyde(MDA) and activity of superoxide dismutase(SOD) and glutathione peroxidase(GSH-PX) in the serum,liver and heart of the mice were tested.Results The SOD and GSH-Px activities in aged model group were significantly decreased and MDA significantly increased compared with normal mice;CPH in different doses evidently increased the SOD and GSH-Px activities and reduced the MDA level in serum,liver and heart in mice.Conclusion CPH can obviously increase the antioxidant ability of mice and has certain anti-aging effects,especially the high dose .