Determination Of Iron Deposition In Liver Fibrosis By Confocal Microbeam X-ray Fluorescence

Part ⅠConfocal microbeam X-ray fluorescence detection of hepatic iron deposition in rat liver fibrosisObjective:Hepatic fibrosis model of rats was established by injection of thiocaracetamide,and the content and distribution of iron in liver tissues at different time points during hepatic fibrosis were determined by confocal microbeam X-ray fluorescence technique.Methods:Fourteen 2-month-old SD rats were selected during the experiment.All rats required for the experiment were divided into control group and experimental group,with 7 rats in each group.Rats in the experimental group were continuously injected intraperitoneally with 4%thioacetamide solution to build a liver fibrosis model,while equal doses of saline were given to all rats in the control portion of the experiment.From the 5th week,rats in both groups were injected with 4%thiocaracetamide solution or normal saline,and the same dose of phosphate buffer solution was added once a week.We will timely monitor the weight changes of the two groups of all rats during the injection process.In both groups,one rat was sacrificed at 0 h,7 d,14 d,21 d,28 d,42 d and 56 d after injection of thioacetamide or normal saline,and part of the liver tissue of each rat was cut for confocal microbeam X-ray fluorescence detection.Part of the liver tissues of the two groups were stained with hematoxylin and eosin on day 56 to detect liver fibrosis.Results:At day 56,hematoxylin-eosin staining results showed no fibrosis in the liver tissues of control group,while there was fibro-collagen formation in the liver tissue of the experimental rats at day 56.Confocal microbeam X-ray fluorescence spectrogram showed that there were iron,potassium,zinc and other metal trace elements in rat liver tissue,and iron was significantly higher than the other two elements.In addition,confocal microbeam X-ray fluorescence detection in the experimental group showed that iron deposition in rat liver tissue gradually increased with the increase of thioacetamide injection times.Conclusion:Injection of thioacetamide reagent into SD rats can lead to the formation of liver fibrosis.Confocal microbeam X-ray fluorescence can detect several different elements in biological samples and can be used to semi-quantitatively evaluate iron deposition in rat liver fibrosis.Iron deposition was positively correlated with liver injury in rat liver fibrosis induced by multiple intraperitoneal administration(thiocaracetamide reagent).Confocal microbeam X-ray fluorescence technique can visually and clearly display the two-dimensional distribution of iron in liver tissue.Part ⅡConfocal microbeam X-ray fluorescence was used to evaluate the value of bone marrow mesenchymal stem cells in alleviating liver iron deposition during liver fibrosisObjective:In the course of hepatic fibrosis induced by thioacetamide,mesenchymal stem cells obtained from bone marrow of rats injected with tail vein,and the confocal microbeam X-ray fluorescence technique was used to evaluate the effect of bone marrow mesenchyml stem cells on reducing liver iron deposition and alleviating liver fibrosis.Methods:To isolate,extract and amplify bone marrow mesenchymal stem cells from healthy rats and evaluate the biological characteristics of the extracted cells.Seven 2-month-old SD rats were randomly selected then,and all rats were injected with thioacetamide for 8 consecutive weeks.Bone marrow mesenchyml stem cells were injected into the tail vein of the remaining rats after thioacetamide injection from the 5th week.During the whole process of drug administration,the changes in body weight of rats in each group were monitored timely.One rat was dissected at 0 h,7 d,14 d,21 d,28 d,42 d,and 56 d after injection of thioacetamide.Part of the liver tissue of each rat was selected for confocal microbeam X-ray fluorescence detection,and hematoxylin-eosin staining of liver tissues of rats on day 56 was used to observe fibrosis.Results:Hematoxylin-eosin staining demonstrated that comparison with thioacetamide only rats in the first part experiment,collagen fibers in the liver tissue of bone marrow derived mesenchyml stem cells were notably reduced,and the liver fibrosis was obviously relieved.Confocal microbeam X-ray fluorescence spectrogram showed that iron deposition in liver still showed an increasing trend from 0 h to 28 d,but iron deposition in liver showed a significant downward trend from 42 d to 56 d.Conclusion:Confocal microbeam X-ray fluorescence analysis can effectively detect the changes of liver iron deposition during liver fibrosis.Hematoxylin-eosin staining and confocal microbeam X-ray fluorescence detection jointly proved that rat bone marrow mesenchymal stem cells can effectively reduce hepatic iron deposition in the process of hepatic fibrosis,repair hepatic damage to a certain extent,and alleviate hepatic fibrosis.