| Iron deposition in liver fibrosis is a frequently observed pathological change.Our previous research had revealed that there was serious iron deposition in the DMN induced rat liver fibrosis.The iron mainly deposited in the kupffer cells.So these researches raised the question that What kind of changes in the kupffer cells was induced by the iron deposition? Is iron deposition can enhance the secretion of liver fibrosis promoting cytokines? How Chinese herb affect the expression of liver fibrosis promoting cytokines induced by iron deposition? This research mainly concentrated on these questions.1Explorartion of the iron concentration for iron depositionA fitful iron concentration is necessarily to be explored before conduct the experiments related to iron deposition in the RAW264.7 cell line for no knowledge about iron deposited in this kind of cell line.A nontoxic concentration of iron must be found before doing of next step of experiment.The results of MTT experiments showed that up to 100uM of ferric ammonium citrate(FAC)of iron has no marked inhibitory effect to the RAW264.7 cell line.Additionally,Prussian blue staining results showed that iron was deposited significantly in the RAW264.7 cells after 24h of FAC interaction in FAC concentration of 100uM.2The changes of liver fibrosis related cytokines mRNA expression after iron deposited in phagocytes.The FAC was added to the cells for 24h and then the cells were collected for RNA extraction and quantitative PCR.The mRNA expression of TGFβ1,PDGF-β,iNOS,TNF-α,MMP-13 were tested through real-time quantitative PCR.The results showed that when compared with blank control cells,TGFβ1 mRNA was significantly down-regulated,PDGF-β,MMP-13mRNA have no marked changes and iNOSmRNA,TNF-αmRNA were up-regulated significantly.3GdCL3,Deferoxamine affect the expression of liver fibrosis related cytokines mRNA in the RAW264.7 cell line with iron deposition.GdCL3,a reagent can block the function of kupffer cells and Deferoxamine,a kind of medicine can relieve the extent of iron deposition. How these two reagents affect the expression of liver fibrosis related cytokines mRNA in the RAW264.7 cell line with iron deposition? The results showed that GdCL3 in concentrations of 10mg/ml,2mg/ml,0.4mg/ml can markedly up-regulated the TGF-β1 mRNA expression in RAW2654.7 cell line with iron deposition.The iNOS mRNA expression was down-regulated by GdCL3 in concentrations of 10mg/ml,2mg/ml,0.4mg/ml,8Oug/ml,16ug/ml.The TNFαmRNA expression was down-regulated by GdCL3 in concentrations of 10mg/ml and 2mg/ml.These results revealed that the phagocyte function of RAW264.7 cell line was blocked by GdCL3.iNOS and TNF-αare proinflammatory factors,down-regulated by GdCL3,that support evidences for the opinion of GdCL3 can reverse liver fibrosis.Similar results were showed in Deferoxamine added to RAW264.7 cell line with iron deposition. These results revealed that therapy of relieving the iron deposition in kupffer cells may facilitate curing liver fibrosis.4Chinese herb complex affect the expression of liver fibrosis related cytokines mRNA in the RAW264.7 cell line with iron deposition.Previous research showed that the expression of liver fibrosis related cytokines mRNA in RAW264.7 cell line can be affected by iron deposition.So what kind of effect can be exerted to the RAW264.7 cell line by a Chinese herb complex? Is Chinese herb complex curing fibrosis through affecting the liver fibrosis related cytokines expressed by kupffer cells?In this research,iron was deposited to the RAW264.7 cell line.The liver fibrosis related cytokines mRNA in iron deposited RAW264.7 cell line affected by a Chinese herb complex were observed.The results showed that the Chinese herb complex can markedly up-regulated the TGF-β1 mRNA expression in RAW264.7 cell line with iron deposition.iNOS and TNF-αmRNA were being down-regulated significantly by the Chinese herb complex.These results revealed that Chinese herb complex Curing fibrosis may through affecting the liver fibrosis related cytokines expressed by kupffer cells.5 Chinese herb complex affect the expression of liver fibrosis related cytokines mRNA in liver tissues of DMN induced liver fibrosis rats.Previous research showed that the expression of liver fibrosis related cytokines mRNA in RAW264.7 cell line can be affected by iron deposition.Chinese herb can improve the expression of the cytokines mRNA significantly.However,are there similar effects in the DMN induced liver fibrosis rats by Chinese herb complex? This research may explore the effects in DMN induced model rats exerted by the Chinese herb complex.The results showed that the expression of TGF-β1mRNA in DMN induced liver fibrosis was opposite to the RAW264.7 cell line,namely the TGFβ1mRNA in DMN induced liver fibrosis was up-regulated.This result revealed that there are differences in DMN induced liver fibrosis rats and in RAW264.7 cell line.However,the expression of iNOS and TNFαmRNA have similar results in DMN induced liver fibrosis rats and in RAW264.7 Cell line.Both the iNOS and TNF-αare important proinflammatory factors which can enhance the development of inflammation. The inflammation must strengthen the development of liver fibrosis.So the iNOS and TNF-αalso can facilitate liver fibrosis development.Additionally,GdCL3 was injected to DMN induced liver fibrosis rats and results showed that the pathological changes in GdCL3 injected rat liver tissues are relieved significantly compared with model group.These results were similar to previous results.In this experiment,the Chinese herb complex was feeded to liver fibrosis model rats for S weeks.The results showed that the Chinese herb complex not only improved the pathological Changes,but also improved the expression of the three cytokines mRNA related to the iron deposition.
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