Dissertation > Excellent graduate degree dissertation topics show

Inflammatory Cytokines Exacerbate Lipid Accumulation and Injury in Primary Hepatocytes Via Up-regulating Srebp-1

Author: BaiLingLing
Tutor: RuanXiongZhong
School: Chongqing Medical University
Course: Clinical Laboratory Science
Keywords: primary hepatocytes Isolation CollagenaseInflammatory cytokines Primary hepatocytes Oxidative stress ER stressInflammation cytokine SREBP-1 ACC alpha FAS
CLC: R575.5
Type: Master's thesis
Year: 2013
Downloads: 62
Quote: 0
Read: Download Dissertation

Abstract


PART ONE:THE ISOLATION AND CULTURE METHOD OFMOUSE PRIMARY HEPATOCYTESObjectives: Liver is a very essential organ for lipid metabolism. Theisolation and culture method of mouse primary hepatocyes was estabishedfor subsequent lipid metabolism of liver.Methods: The mouse primary hepatocytes were isolated by a two-stepperfusion with type IV collagenase in situ. The trypan blue staining wasused to detect cells viability. The attachment of primary hepatocytes wasobserved at different time points. Albumin in supernatant which wassecreted by primary hepateocytes had been detected at different time pointsfor a week. Intracellular Albumin and glycogen was examined byimmunocytochemistry staining and PAS staining for the identification ofhepatocytes.Results: trypan blue results show: the highest cells viability was up to90%. The fresh isolated cells are almost alone and stereo. after4hours, most of hepatocytes have been attached. A typical dual-nucleus structure and flatcytoplasm were observed in hepatocytes. After1day, dual-nucleus structurewas more obvious and nucleoluses were clearly visible and cytoplasm wasmore flat. after2days,cell cytoplasm was turn into polygon, and maintainthis form for several days. Almost all of the isolated cells were primaryhepatocytes by the identification of PAS staining andimmunocytochemistry staining for glycogen and albumin. Albumin haddeferent levels in supernatant at different time points in a week.Conclusion: primary hepatocytes from mouse were successful isolatedand cultured and could be used for subsequent research in a week. PART TWO: EFFECTS OF INFLAMMATION CYTOKINES ONLIPID ACCUMULATION AND INJURY IN PRIMARYHEPATOCYITESObjectives: in recent years, chronic systemic inflammation has a veryimportant role in the metabolic disease. In this study we will discuss whetherinflammatory cytokines increase TG and FFA levels and cause damage inmouse primary hepatocytes.Methods: The cells were treated with inflammatory cytokines (TNF-α or IL-6) in the absence or presence of palmitic acid (PA). Lipid accumulationwas observed by Oil red O staining and quantitative detection of intracellularTG and FFA. Oxidative stress status was determined by H2O2and ROS tests.ER stress was detected by IRE1, ATF6, GRP78mRNA levels.Results: Oil Red O staining and quantitative Measurement of TG andFFA demonstrated that inflammatory stress increased lipid accumulation inthe primary hepatic cells with or without palmitic acid loading. Results alsoshowed that inflammatory cytokines can increase H2O2and ROS levels andIRE1, ATF6, GRP78gene expression levels.Conclusion: inflammatory cytokines cause lipid accumulation anddamage in mouse primary hepatocytes. PART THREE: THE EXPRESSION OF SREBP-1AND ITSDOWNSTREAM TARGET GENGE IN PRIMARY HEPATOCYTESUNDER INFLAMMTORY STRESSObjective: SREBP-1, a nuclear transcription factor, which is abundantin liver, regulates the synthesis of fatty acids and triglyceride in the liver viaregulating the expression of its downstream target genes ACC alpha and FAS. Our study is to observe the expression of SREBP-1and ACC alpha andFAS in primary hepatocytes under inflammatory stress.Methods: The mouse primary hepatocytes were treated withinflammatory cytokines (TNF-α or IL-6) in the absence or presence ofpalmitic acid (PA).The mRNA and protein expression of SREBP-1and ACCalpha and FAS in primary hepatocytes were determined by real-time PCRand Western blotting.Results:Our data shows that Inflammatory cytokines can up-regulatethe mRNA and protein expression of SREBP-1and ACC alpha and FAS inmouse primary hepatocytes with or without palmitic acid loading.Conclusion:the expression of SREBP-1and ACC alpha and FAS in mouseprimary hepatocytes can increase under inflammatory stress.

Related Dissertations

  1. Isolation & Characterization of a Cetochlor Degrading Strain Y3B-1 and Its Degrading Characteristics,X172
  2. Studies on Isolation, Purification and Chemical Structure for Polysaccharides from Camellia Chrysantha(Hu) Tuyama,S567.19
  3. Functional Analysis of tal (transcription activator-like) Genes of Xanthomonas Oryzae,S435.11
  4. Purification and identification of tobacco materials alkaloids,S572
  5. Isolation and Characterization of Carbendazim-Degrading Strains, Cloning and Expression of the mheI Gene,X172
  6. Isolation and Identification of Avian Pathogenic Escherichia Coli from Chicken and Characterizations of Its Virulence-Associated Genes,S852.61
  7. Isolation and Identification of Infectious Bronchitis Virus and Sequence Analysis of Its S1 Gene and N Gene,S852.65
  8. Isolationand Identification of Porcine Parvovirus and Parts of Its Biological Characteristics,S852.65
  9. Characterization of an Atrazine-Degrading Strain, Cloning of Key Degrading Related Genes and Construction of a Gene Cluster,X172
  10. Lipase-catalyzed Transesterification of Lard to Produce L-ascorbyl Fatty Acid Esters,TS221
  11. Study on Antioxidant Activities and Lipase-catalyzed Transesterification of Lard to Produce L-ascorbyl Fatty Acid Esters,TS202.3
  12. Functional Analysis of Tal(Transcription Activator-like) Genes of Xanthomonas Oryzae,S435.111.4
  13. Study on Technique of Protein and Starch Extracted from Wheat Bran,S512.1
  14. Isollation and Identification of Haemophilus Parasuis and Its Proteomic Analysis under in Vitro Growth Conditions Mimicking the in Vivo Environment,S858.28
  15. Isolation and Identification of a Probiotic Strain Capable of Producing Glycerol and Optimization of Fermentation Conditions for the Strain,S823.5
  16. The Research on the Antioxidant Fat-Solube Compositions of Salicornia Herbacea L.,R284.1
  17. The Investigation on the Renal Injury and Effects on Oxidative Stress of Guinea Pigs Induced by Trichloroethylene,R363
  18. Isolation and Identification of Photosynthetic Bacteria and Preliminary Study of Proteinase Secreted from Rubrivivax Gelationsus,Q93
  19. Isolation and Identification of Antimicrobial Substance Produced by Bacillus Subtilis fmbJ,TQ920.6
  20. Heme Oxygenase-1 is Involved in the Modulatory Role of Salicylic Acid in Cadmium-Induced Oxidative Stress in Medicago Sativa,S541.9
  21. Effects of Cumulus Cells on Maturarion in Vitro and Subsequent Developmental Competence of Porcine Oocytes,S828

CLC: > Medicine, health > Internal Medicine > Digestive and abdominal diseases > Liver and gall bladder disease > Liver metabolic disorders
© 2012 www.DissertationTopic.Net  Mobile