| Introductionliver cirrhosis is a disease which threatens the human' s life and health seriously. The process of liver fibrosis which are caused by various factors is always associated with the increase and abnormal distribution of interstitial colla-gens and other matrix components, therefore, the research of anti - fibrosis becomes the key link to prevent and cure the cirrhosis and portal hypertension. Previous studies show that an essential section of liver fibrosis is the formation of myofibroblast ( MFB) which is transited from hepatic stellate cell ( HSC) in the inflammation and necrosis area of the injured liver. The successful vitro culture of HSC provides an ideal cell model for the investment of liver fibrosis. In the middle of 1980s, recombined interferon gamma (IFN -γ ) was created through gene engineering technology, and it has been applied to clinic extensively since then. Recently, reports abroad show IFN - 7 have strong function of anti - fibrosis which is based on the animal and vitro experiments . The purpose of this research is to present IFN - γ s function mechanisms of the anti - fibrosis through observing the influence of IFN -γ on the vitro HSC' s extracellular matrix ( ECM ) secretion, and to provide further theoretical foundations for treatment of liver fibrosis in clinic.MaterialCell line:Hepatic stellate cell (rHSC -99)Main Reagents: Hyaluronic Acid RIA Kit, Laminin RIA Kit, Procollagen Type I RIA Kit, DMEM, Fetal Bivine Serum, 25cm~2 culture flask, 96 -wellculture plate , Recombinant Human Interferonγ for Injection, other daily medicines are offered by center laboratory and immune laboratory .Main instruments: Inverted Microscope, GC - 1200 Gamma Radioimmuno-assay Counter, KDC -2046 Low Speed Refrigerater Centrifuge, Nu 4500E CO—2 Water - Jacketed Incubator, MDF - U4086S deep low -temperature refrigerator, MDF-U0541 low -temperature refrigerator, GB11241 -89 electric heating constant temperature water tank, ZBY149 - 83 electric heating constant temperature drum wind drying oven。MethodRefer to serum - free raise. Levels of PCIII, LN and HA were examined by radiommunoassy ( RIA ).Statistic Analysis All data were expressed as the mean ± SD. SPSS11. 5 statistic software is applied. ONE - WAY ANOVA was used to compare differences. Regression analysis was used to evaluate the correlation between variables. All statistical tests were two - tailed and p <0.05 was regarded as statistically significant.ResultsI The influence of IFN -γ on rHSC - 99 ' s shape and growthUnder levels of dose of IFN — γ, rHSC - 99 grows well and its shape does not change obviously.II The influence of IFN -γ on rHSC -99 ' s ability to secrete PCIII,LN, HAThe impact of IFN - γ on the secretions of PC III ,LN,HA will be stronger with a higher dose. Except for the case of several lower dose IFN - 7 treat group, under the same culturing time there are significant difference between IFN - γ treat group and blank comparison group( P <0.05,P <0.01). IFN - 7 significantly reduces PC III, LN, HA secreting when the culturing time is prolonged. There are no significant difference between 48 hours of culturing and 24hours of culturing under the same dose (P > 0. 05). However, a significant difference is presented in the case of 72 hours compared with the case of 24 hours ( P < 0. 05 ). Except for HA, the levels of PC III and LN are -negatively correlated with the dose and culturing time of IFN - γ.DiscussionHSC has the ability to secrete ECM and cytokines, and can contract. It is known that HSC plays main role in the development of the cirrhosis and portal hypertension. rHSC -99 is an immortalized rat HSC line and an ideal cell model in the research of anti - fibrosis. IFN - γ is a lymphokine produced by T lymphocytes and natural killer cells during the process of immune response. Recently , many researchers verified IFN - γ is regarded as a desirable anti - fibrosis medicine.Additionally, it is show that IFN - γ has a suppression effect on the secretion of ECM from rHSC - 99 when the concentration of IFN - γ' s is between 100U/ml and 1000U/ml, and the effect is dependent on the dose and the culturing time. This conclusion is consistent with the results of animal experiments and vitro experiments in the past. The last result shows that the effect of the IFN -γ on the secretion of ECM by rHSC - 99 is not statistically significant difference between the treat group 500 U/ml and 1000 U/ml. This may indicate the existence of the quantity - effect platform stage. This research shows that IFN -γ can suppress the cell rHSC - 99 to secrete ECM , and play an anti - fibrosis role. Therefore IFN - γ can be applied to the clinical generally.ConclusionI There is no obvious effect on rHSC - 99' s shape under any dose of IFN -γ.II IFN - γ has a negative effect on the secretion of ECM by rHSC - 99, and exists concentration - effect relationship and time - effect relationship.Iff The present study further verify IFN - γ have the function of anti - fibro-...
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