| Aging is the ongoing change process.Theory of Free radicals is one of the most representative theory of aging,It was assumed that the free radical damage human cells, and then cause various diseases, so as to accelerate the aging process.Oxidation is one of the important steps to prevent aging, antioxidants were any exist in low concentration could effectively restrain free radical oxidation reaction of material, development of practical,high efficiency, low toxicity of natural antioxidant is the focus of future research.Chinese traditional medicine, formerly known as fairy housefly maggot, commonly known as maggots, its adult housefly haunt filthy place, place oneself in a countless number of bacteria, but can be safe and sound, don’t be misled by the pathogen infection, this is the result of its excellent immune function.In recent years, domestic and foreign researchers in the study of ROM fairy, focused on the active peptide, they found that the ROM fairy extract rich in lectins, lysozyme, active ingredients, such as antibacterial protein, have fight bacteria, fungi, viruses and other pathogens and resistance of tumor cells.With the deepening of the research, some studies also found that the ROM fairy extract can remove free radicals and resistance to heavy metal stress.The lab early in the fairy extract on hyperlipemia rats blood lipid regulation and the role of liver protection, fairy extract to liver homogenate Chinese SOD activity increased significantly, MDA level declined obviously,prompt fairy extract might have by scavenging free radicals to enhance the effect of antioxidant capacity.This research use D-gal caused by aging and natural aging mice model at the beginning of the rat model of ROM fairy extract anti-aging efficacy of comprehensive evaluation, and its antioxidant mechanism were discussed, on the basis of the digestion process of early ROM fairy extract has carried on the preliminary study, is developed from the traditional Chinese medicine luo fairy an anti-aging candidate drug molecules to provide theoretical basis, also for the extraction and separation of the ROM fairy antioxidant peptides to lay the foundation.Methods1. The use of D-gal induced mouse model to evaluate early ROM fairy extract anti-aging effects and oxidation mechanism48, kunming mice were randomly divided into normal control group, aging model group,Vit E, the control group and early ROM fairy extract low, medium and high dose groups,each group of eight.In addition to the normal control group, the rest of the group are calculated by 10 mg/kg for D-gal neck back subcutaneous injection, once per day, injection of 6 weeks in a row, at the same time early ROM fairy extract and Vit E group in 10 ml/kg lavage.Building to 0 d, determination of mice weight every day, and watch the groups of mice morphological change;Building for 30 d, to avoid dark experiment in mice;And in building 6 weeks after the death mice, detection in serum, liver and brain tissue antioxidant related indicators.2. The use of natural aging rats model evaluation, at the beginning of fairy extract anti-aging efficacy and its antioxidant mechanism10 to 11 months in SPF rats environment raising nine months, raising to 19-20 months of age, were randomly divided into 5 groups, respectively, the aging model group, Vit E and early ROM fairy extract low, medium and high dose groups, each group of eight.Youth in the control group in aged model group rats building and approaches to medicine for 10 days before the date of purchase, adaptive breeding for 10 days.ROM fairy extract at the beginning of each dose group and Vit E group by 10 ml/kg, to fill the stomach, once per day, 2 consecutive months.Young and aged model group were treated with the volume of normal saline.Building to 0 d, monthly determination of rat body weight, and observe each group rats morphology change;After Morris water maze experiment, rats injected ether anesthesia, abdominal aorta in blood;Quickly remove the brain tissue after taking blood.3. 3. Preliminary digestion early fairy extract Early ROM fairy extract to make use of alkaline protease preliminary digestion, HPLC analysis component differences before and after enzymolysis, and determine its clearing DPPH· free radicals activity;On the basis of the single factor analysis is adopted to p H,temperature, substrate concentration, enzyme amount and enzymolysis time and so on enzymatic hydrolysis conditions were studied.Results1.1.Anti-aging effect of Luoxianzi extract on D-gal induced aging model mice Compared with normal control group, for aging model mice, the weight increasement declined. The number of errors in step-down test increased, the SOD and GSH-PX activity in serum, liver and brain tissues dropped and the MDA content was raised. Compared with model group, for the micein luoxianzi extract and Vit E treatment groups, the weight increasement rised. the SOD and GSH-PX activity in serum, liver and brain tissues enhanced and the MDA content reduced.2.Anti-aging effect of Luoxianzi extract on natural aging rats Compared with the young control group, aging model group rats activity is slow, the skin filth matte, weight with the increase of the weeks is not obvious;Morris water maze is directional navigation experiments of potential learning and memory dysfunction;Rats tissue paraffin section of HE staining results and ultrastructure of tem results show that the cytoplasm of neurons, the cell body and bumps in lysosome;Cell mitochondria swelling in the body, and the dissolution of cristae in mitochondria disappear;Rough endoplasmic reticulum expansion, the expansion of the net cavity, free ribosome density decreased,decrease in the number, distribution, irregular arrangement, distortion and inflation, and some free in the cytoplasm;Neural synaptic cleft and presynaptic membrane structure after,fuzzy, neural synaptic connections less;The brain activity of SOD and gsh-px activity,MDA content increased.Compared with aging model group, Vit E and early ROM fairy extract each dose groups with weeks of weight gain, reduce the learning and memory dysfunction, rats tissue paraffin section of HE staining results and ultrastructure of tem results show that the cytoplasm of neurons, the cell body and bumps in the decrease in the number of lysosome;Mitochondrial swelling in the cell body is abate, mitochondrial crest in the dissolved situation has eased;Neural synaptic cleft and presynaptic membrane structure after, is clear;In brain tissue SOD activity increases, reduce MDA content and increase gsh-px activity.3.Preliminary enzymatic hydrolysis of Luo Xianzi This study about the fairy after enzymolysis, with early extraction by HPLC(High Performance Liquid Chromatography, HPLC) in sample analysis and determination of the removal of DPPH· found that before and after enzymolysis of fairy the extract Chromatography graph at the beginning of the differences, and after enzymolysis ROM fairy extract has better removal at the beginning of the activity of DPPH ·.Through subsequent preliminary optimize the conditions of enzymatic hydrolysis, enzymatic hydrolysis in the substrate concentration, enzyme solution p H and enzyme solution temperature on the influence on the efficiency of the enzyme solution, but the amount of enzyme and enzyme solution time have less effect on the efficiency of the enzyme solution.Conclusion This study used respectively D-gal to subacute aging model mice and natural aging rats model of comprehensive evaluation at the beginning of the ROM fairy extract anti-aging efficacy, the experimental results show that the D-gal to subacute aging model mice and natural aging rats compared with normal control group, the symptoms of aging.And luo fairy experimental animals of every dose group and Vit E aging symptoms are mitigated.By observing each experimental animal behavior, determination of antioxidant in vivo related indicators, detects the ultrastructure, the analysis found early ROM fairy extract may through scavenging free radicals and other role to enhance the antioxidant capacity of the body, and then improve the ultrastructure of some of the symptoms of aging, so as to achieve the role of anti-aging.About fairy after initial enzyme solution, with early extraction of sample by HPLC in the analysis and determination of its clearing DPPH·free radicals activity found that the enzyme solution before and after the differences between early ROM fairy extract chromatography pictures, show luo at the beginning of fairy extract after enzymolysis, hydrolysis does a certain antioxidant activity of the material.Through subsequent preliminary optimization of enzymatic hydrolysis conditions found that enzyme substrate concentration, enzyme solution p H and enzyme solution temperature on the efficiency of the enzyme solution, but the amount of enzyme and enzyme solution time have less effect on the efficiency of the enzyme solution.The results from traditional Chinese medicine for ROM fairy developed an anti-aging candidate drug molecules provide the theoretical basis, also for the extraction and separation of the ROM fairy antioxidant peptides to lay the foundation. |
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