Over-expression Of C/EBPα In PARs Activated HSC And Its Negative Regulatory Role In The For Mation Of Liver Fibrosis

Liver fibrosis is one of the serious disease that threats human health, and so far noideal treatment method is available. Most scholars from domestic and overseas believe thathepatic stellate cells （HSC） activation is the key of liver fibrosis, so inhibiting HSCactivation plays important role in liver fibrosis preventive treatment.Protease activated receptors （PARs） belongs to the G-protein coupling receptor familymembers. It participates in the process of cell proliferation and differentiation regulation,and related with varieties of pathological physiology process. In our prophase researchconcerns PAR_1、2regulat HSC activation and proliferation show that PAR_1、2is the maininflammation and fibrosis receptors in the process of the liver fibrosis automatically cyclicamplification which mediated by blood coagulation enzyme and trypsin, and its expressioncan not only promotes the formation of liver fibrosis, also aggravates liver inflammationcascade, plays positively stimulative role in the form of liver fibrosis. HSC inhibition,collagen production blocking, liver inflammation limitation, and interruption ofautomatically cyclic amplification function subsequent to the HSC activation in the processof liver fibrosis can be achieved by antagonizing PAR_1、2activity. But so far effectiveselectivity PAR_1、2inhibitors and methods has not been found in the liver fibrosis preventivetreatment research.It is found in liver fibrosis study that significant changes in activated HSC are lipiddroplet cells decreased or even disappeared. Lipid droplet disappearance means de-fat celldifferentiation and HSC is necessary for maintaining the de-fat cell static state, so de-fatcell differentiation plays a key effect in HSC activition. The expression of CCAATenhancer binding proteins a （C/EBPa） genes is one of the important transcription factors tomaintain adipose cell differentiation, and the main function is to promote adipose cell intoterminal differentiation, enhanced separate C/EBP α expression is enough to inducedifferentiation from preadipocyte3T3fat cells to adipose cell. C/EBPα expression in liver tissue cells and static HSC is confirmed and negative expression in the active HSC. Thenwhether HSC static can be maintained by C/EBP α related with Lipid droplet formation andthe adipose cell differentiation, is there any negative regulatory role for PARs to promoteliver fibrosis by introducing C/EBP α to maintain adipose cell differentiation andpreventing HSC activation mechanisms?Consider of the previous cognition, on the basis of PAR_1、2activate HSC and promotepositive regulatory in liver fibrosis, to construct C/EBP α carrier and transfection through ofPAR_1、2-activated HSC,observe the function of antagonism of PAR_1、2and upregulation ofC/EBPαexpression in C/EBPα in negative regulatory mechanism of HSC activation,provide the basis for the prevention and treatment of liver fibosis.The main results and conclusion of the study:1、The original cytology HSCs characteristics are kept in HSC-T6such as star or flatspindle appearance, also activated HSC-T6expresses α-SMA and synthetic type I,IIIcollagen, so the invivo biology process can be well reflected. It is the ideal cell model forliver fibrosis study.2、PcDNA3.1（-）-C/EBP α plasmid recombinant is appraise with double digestion andcan be completely digested back to the molecular weight size of C/EBP α and pcDNA3.1（-）carrier, which shows that the plasmid construction is successful, and there is goodhomologue between the plasmid recombinant and cultured HSC-T6.3、HSC-T6can be effectively transfect by Exogenous C/EBP α and highly expressedafterwards. In some extent, HSC-T6activation and collagen synthesis can be inhibited byraising C/EBP α expression.4、When exogenous C/EBP α is introduced to HSC-T6cells activated by PAR_1、2, theα-SMA and type I procollagen expression decreases, meanwhile, cell proliferation degreealso reduces. It is suggested that in some extent raised C/EBP α expression inhibitspromotive role of PAR_1、2to HSC activation, proliferation and secrete collagen.