Dissertation > Excellent graduate degree dissertation topics show

Experimental Study of Immunoregulation and Antitumor Function of Gekko Gecko Peptides

Author: XiZuo
Tutor: XieYuAn
School: Guangxi Medical University
Course: Oncology
Keywords: Gecko Protein Separation and purification Gecko peptide In vitro Immune function MTS assay Immunocompromised MTS assay Tumor-bearing mice Immunomodulatory Antitumor
CLC: R285.5
Type: Master's thesis
Year: 2011
Downloads: 93
Quote: 0
Read: Download Dissertation

Abstract


Objective: purified protein peptides isolated from Gecko and Determination of protein molecular weight, and component concentration. Methods: fresh Gecko sacrificed to the head and viscera, cut into small tissue blocks, weighing to 0.3% NaCl solution by 5ml / g diluted pulverized homogenized, liquid nitrogen freezing and thawing cycle is repeated, 4 ℃, 10000r/min low-temperature high-speed centrifugal 30min, repeated freezing and thawing three times repeat the operation once. The resulting supernatant was filtered through filter paper to remove insoluble lipids 0.45 UM filter filtering then with closure molecular mass of 50KDa ultrafiltration centrifuge tube was subjected to ultrafiltration, macromolecular substance is removed, and the resulting filtrate is freeze-dried to give a brown solid powder was dissolved after protein concentration was measured by Sephadex G-50 gel filtration chromatography, SDS-PAGE electrophoresis analysis identified protein purity and determination of molecular weight, UV spectrophotometer. Results: the Gecko protein by Sephadex G-50 column eluting three major protein peaks by SDS-PAGE electrophoresis analysis Gecko peptide fractions I molecular weight between 20Kda-50Kda component Ⅱ molecular weight in between 3Kda-20Kda, Gecko peptide the molecular weight of the fraction Ⅲ 3 kDa or less, and the UV spectrophotometer component concentrations respectively of the three groups of proteins 0.32mg/ml, 1.28mg/ml, 0.25 mg / mL, the collected amount respectively 140ml, 161ml, 147ml, protein contents were the 44.8mg, 206.08mg, 126.42mg, a total of 377.3mg, accounting for 62.88% of the Gecko frozen ten protein powder, three groups Gejie peptide components respectively 7.47%, 34.34%, 21.07% of the total protein. The Conclusion: Gecko homogenate by ultrafiltration and centrifugation, freeze-drying and gel chromatography method can be isolated and purified protein of molecular weight to 50Kda-3Kda the. Objective: To study the Gecko peptide in vitro immune function, filter out the most active component the Gecko peptide immune. Methods: The preparation of different concentrations (0.1ug/ml, 1ug/ml to 10ug/ml, 100ug/ml) a Tokay peptide solution and different concentrations (10U/ml, 100U/ml, 1000U/ml, 10000U/ml) IL-2 solution, different concentrations of the Gecko peptide in vitro study different components alone or combined with IL-2 Application of immune cells, MTS Determination of NK cell activity and macrophage tumoricidal activity. Results: Gecko peptide fractions II 3 concentration (1ug/ml, 10ug/ml, 100ug/ml) can significantly increase NK cell killing activity (P lt; 0.01) in the concentration 10ug/ml, NK cell activity up enhance the role of (P gt; 0.05) (17.26 ± 3.07)%, 1.58 times more than the normal control group, but the concentration of 0.1 ug / ml, there was no Gecko peptide fractions Ⅰ Ⅲ alone, there was no increase NK cell activity. Gecko peptide fractions II, the use of the different component concentration of a 10ug/ml the Gecko peptide with different concentrations (10U/ml, 100U/ml, 1000U/ml, 10000U/ml) of IL-2 mixed with IL-2 has a synergistic effect (P lt; 0.05); When different concentrations of Gecko peptide with 1000U/ml IL-2 is mixed 10ug/ml of Gecko peptide fractions II IL-2 activated NK cells can be enhanced role (P LT; 0.01) . Gecko peptide three components can enhance macrophage tumoricidal activity, wherein the component Ⅰ, Ⅲ lug / ml concentration 10ug/ml when 100ug/ml enhance the role of significant (P lt; 0.05); component Ⅱ 4 concentration can significantly enhance macrophage tumoricidal activity, macrophage tumoricidal activity concentration lOug / ml, respectively, up to (31.99 ± 6.97)%, compared to the normal control group (15.59 ± 1.63)% 2.05 times respectively, to enhance the role of significant (P lt; 0.05). Gecko peptide fractions Ⅰ, Ⅲ concentration 0.1ug/ml has enhanced the role of the macrophage tumoricidal activity. Conclusion: the Gecko peptide collected three major components, component II vitro application on the NK cell activity to enhance the role to enhance the strongest effect when the concentration of 10ug/ml; and at the concentration of 10ug/ml collaborative IL-2-activated NK cells. Gecko peptide fractions Ⅰ Ⅲ alone, there was no increase NK cell activity. Gecko peptide three components in vitro applications can enhance macrophage tumoricidal activity, wherein component II 10ug/ml time, enhance the role of the strongest; Gecko peptide fractions I, III In concentration 0.1ug/ml does not have enhance macrophage killing of tumor activity. Tip the the Gecko peptide in vitro applications can increase the activity of immune cells, which activate cellular immunity. Objective: To observe the the Gecko peptide on the immune function of mice immunocompromised model. Method: the experimental separation and filter out the molecular weight for the the Gecko peptide component of 3Kda-20Kda II for animal experiments. Intraperitoneal injection of cyclophosphamide (CTX) 40mg/kg/d, continuous administration 3d, mice immunocompromised model to study the the Gecko peptide gavage treatment of mouse immunosuppression model immune function. Immunodeficient mice Gecko peptide fractions were given Ⅱ 500,200 mg / kg / d for 7 consecutive days gavage treatment with MTS colorimetric determination of the stimulation index of spleen lymphocytes, natural killer cells (NK) activity of peritoneal macrophages cells kill tumor activity. Were given peptide component of immunodeficient mice Gecko II 500,200,100 mg / kg / d for 7 consecutive days gavage treatment to HCso Serum hemolysin content of the Gecko peptide immunocompromised model in mice serum hemolysin. Results: The the immunocompromised model CTX established mice by intraperitoneal injection of immune indicators than the normal group were significantly decreased, and the difference was statistically significant (P lt; 0.01). Molecular weight of 3Kda-20Kda Gecko peptide fractions II 500, 200 mg / kg / d orally treatment can significantly improve the immunocompromised mouse model of spleen lymphocyte stimulation index (P lt; 0.05), NK cell activity (P lt; 0.05) and macrophage tumoricidal activity (P lt; 0.05). Gecko peptide fractions II gavage 500,200,100 mg / kg / d mice immunocompromised model serum hemolysin content (P lt; 0.01, P lt; 0.01, P lt; 0.05) can significantly improve. Conclusion: The molecular weight of the Gecko peptide component 3Kda-20Kda II by gavage Cellular Immune, and improve the level of serum hemolysin, in order to enhance the immune function of the immunocompromised mouse model. Objective: To observe the Gecko peptide immunomodulator S180 and Hepal-6 tumor-bearing mice mice transplanted tumor growth inhibition. Methods: the above experimental separation screened Gecko peptide fractions II molecular weight between 3Kda-20Kda, and this component is used in animal experiments. S180 and Hepal-6 tumor-bearing mice as a model to study the Gecko peptide and its therapeutic effect in combination with cyclophosphamide (CTX), MTS Determination of mouse peritoneal macrophages to kill tumor activity, stimulation index of spleen lymphocytes, natural killer cell (NK) activity, saying the weight of tumor inhibitory rate was calculated. Results: simple Gejie peptide fractions II gavage 400mg/kg/d, for 12d, can significantly enhance the S180 mice peritoneal macrophages tumoricidal activity and Hepal-6 tumor-bearing mice peritoneal macrophage phagocytic function (P lt; 0.05), Gecko peptide fractions II after treatment S180 Hepal-6-bearing tumor mice spleen lymphocyte proliferation and NK cell activity were significantly increased (P lt; 0.05). When combined with CTX, Gecko peptide fractions II enables to improve the immune index inhibited by CTX and can significantly improve tumor inhibition rate. Conclusion: The molecular weight of the Gecko 3Kda-20Kda peptide component of the tumor and chemotherapy drugs caused immune suppression regulation synergistic chemotherapy drug anti-tumor effect and can be reached through this route.

Related Dissertations

  1. Research on Combinatorial Regulation of Multiple Transcription Factors,Q78
  2. Recovery Whey Soy Protein by Microfiltration and Development of Whey Powder,TQ936.2
  3. Research on Disease Related miRNA Mining Based on Protein-Protein Interaction Network,R341
  4. The Research and Implementation of Protein Classification Algorithm on the Basic of String Kernel,TP301.6
  5. Study of Oridonin on SGC-7901 Cell Proliferation by Inhibitting Cell Cycle Proteins,R285
  6. Expression of hBMP4 and hBMP7 in Chinese Hamster Ovary Cells,Q78
  7. The Study of Color Change and Control Technology for Oyster Protein Beverage,TS254.4
  8. The Value of the Heart-Type Fatty Acid Binding Protein in Early Diagnosis of Acute Myocardial Infarction,R542.22
  9. Serum A-FABP Levels and Its Expression in Atherosclerotic Plaques of Diabetes Mellitus,R587.1
  10. Impact on Drosophila Brain Developement Through the Overexpression and Loss of Function of Gsα,Q75
  11. Study on Deodorization Techniques of Oyster Protein Beverage,TS254.4
  12. Preliminary Study of Phosphorylation Effect on UGT1A3 Metabolism Activity Towards Drugs,R346
  13. Study on the Preparation of the Low Molecular Weight Fuciodan from Sargassum Henslowianum (C.Agardh) and Anti-tumor Activity,TS254.9
  14. Study on Preparation and Properties of Protein Isolates from Tilapia,TS254.4
  15. Design, Synthesis and Biological Evaluation of Novel Podophyllotoxin Derivatives as Antitumor Agents,R284
  16. Expression and Purification of Influenza a Virus M2 Protein and Detection of Its Antigenicity,R392
  17. Broiler Responses to Reduced-protein Diets Suppleme Nted with Synthetic Amino Acids and Small Peptides,S831.5
  18. The Study on Anti Tumor and Anti-oxidation Activities in Vitro of Extracts from Oxalis Corniculata L,R285
  19. The Preparation of Hpv58e7 Protein by GST-tagged Prokaryotic Expression System,R392
  20. Mutagenesis of a Laccase Gene from Pleurotus eryngii in Vitro and Expression in Pichia Pastoris,TQ925
  21. Functional Analysis of Proteins Encoded by RNA2 of Wheat Yellow Mosaic Virus,S435.121

CLC: > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology
© 2012 www.DissertationTopic.Net  Mobile