Experimental Study On Protective Effects Of MaFGF On Hepatocytes Damaged By CCL₄ In Vitro

Objective: To investigate the inhibiting effects of CCL₄on hepatocytesand to explore the protective role of MaFGF in the injury of CCL₄onhepatocytes of BRL-3A. To study the protective mechanism of MaFGF,and alsoto provide experimental evidence and the theoretical basis for clinicalapplication.Methods:using BRL-3A cell lines which came from Rattus norvegicus ratfor this experiment,the BRL-3A were adherent growth property cells.BRL-3Awere cultured with 10%FBS and RPMI-1640 medium in well plates and culturebottles. And put in CO2incubator . Subculturing cells were seeded in cultureplates of 96 wells:（1）After 72h culture,different concentrations of MaFGFwere added for 12h, The viability and proliferation were determined by the wayof MTT.（2）After 72h culture, different concentrations of CCL₄were added,and MaFGF with different concentration was added further for 12h. the cellviability was tested by the way of MTT.（3）The injury model was induced byCCL₄. And the protective effect of MaFGF was observed on the injuredhepatocytes. The statistical analysis was executed by SPSS 16.0 software, usingT test and one way analysis of variance for statistical, statistically significantlevel was“a=0.05”.Results:（1）The cultured cells had the typical multi-angular morphologicalfeatures of epithelium, Closely spaced, strongly transparency and refraction, cellcytoplasm is rich and stereo sense is strong.（2）Proliferation activity of MaFGF in hepatocytes：No significant difference was noted between low-concentration（1.56-4.68μg/L） MaFGF group and the control group（P>0.05），while MaFGF≥6.24μg/L,there was significant difference with the control group（P<0.05）,butwhen MaFGF≥9.36μg/L,there was lower Proliferation activity,and there wassignificant difference with the last group（P<0.05）（.3）The inhibitory effect of theCCL₄on hepatocytes increased with the increase of concentration.（4）Thebiochemical and enzyme indexes were significantly different between the CCL₄group and control group （P <0.05）; Compared with CCL₄group，the SOD、GSH-Px activity were significandy higher, but the contents of NO，MDA，LDH，AST，ALT were significantly lower （P <0.05）in CCL₄+MaFGF groups. Theactivity and contents of the biochemical and enzyme in CCL₄+MaFGF groupswere not declined to normal levels, comparing with control group, there was stillsignificant difference （P <0.05）.Conclusion：①MaFGF performed some proliferation on hepatocytes atproper concentrations. And between some concentrations, the activity showedconcentration-effect relationship;â‘¡The injured model of hepatocytes induced by CCL₄was successfully built：CCL₄could inhibit hepatocytes proliferation in vitro ,and showed concentration-dependent relaionship.â‘¢MaFGF showed protective effects on the hepatocyte injured by CCL₄,andeffectively reduce the damage by free radicals to hepatocytes.