| Liver injury is the common pathological changes of all the liver diseases. The particularity of the liver make the liver protection medicines be the most used medicines in liver diseases. Liver protection effect of medicines for liver diseases is the most commonly researched drug effect.There are several kinds of animal models can be used for liver injury research at present. But all these models have limits in some extent. Additionally, most of the models are not easily for conduction and not very stabilization. Better animal model for liver injury was tried to develop in this research. Peritoneal injection of DMN at single time was conducted for modeling liver injury. Then the model was used for evaluation of the liver protection effects of a Chinese herb. 1Exploration of the DMN dosage for liver injury modelingThree dosages of DMN include 30mg/kg,20mg/kg,15mg/kg were injected respectively. Consequently, all mice in the group of 30mg/kg were died between 12—24h. The mice of 20mg/kg were died wholly between 24-48h. However, the mice in the 15mg/kg group only have one died at 48h post DMN injection. There were 5 mice lived until 168h post DMN injection. So the 15mg/kg dosage of DMN injection was selected for liver modeling. 2 The pathological characteristics of DMN injection based liver injury model.After DMN injection, mice were sacrificed and serum and liver tissue samples were collected at time points of 6h,12h,24h,36h,48h,72h,120h post injection respectively. Four mice were sacrificed at each time points. The results showed that the ALT in serum was elevated sharply at 6h post DMN injection and kept high level in 12-48h post modeling. Then the ALT level descended quickly after time point of 48h. The same tendency was observed in AST in serum. The albumin in serum was descended from 36h post modeling. But the albumin ascended at time points of 120h post modeling. The total protein concentration in serum was quickly descended at 6h post modeling and reached lowest level at time point of 36h. Then the total protein concentration ascended slowly. However, the total protein concentration was lower than control group at 120h post modeling. Lobule structure in Liver tissue of normal group is maintained normal and aligned regularly. No bleeding, no denaturation, no necrosis, no fat accumulation was found in the liver tissue.Lobule structure in Liver tissue of mice modeled 6h after showed normal. But vacuolar degeneration was found in mass of liver cells. perifubular cells are normal. Denatured cells are mainly located in tissues between vessels. Apoptotic bodies and bleeding were occurred in denatured tissues.Vacuolar degeneration was deteriorated in 12h post modeling and tendency of bleeding was more outstanding. Big area of bleeding and necrosis occurred in some of liver tissue. Plenty of inflammatory cells infiltrated to the edge of necrosis and bleeding area. Apoptotic bodies and bleeding were occurred in denatured tissues.Big area of bleeding and necrosis occurred in most liver tissues at 24h post modeling. Lobule structure of necrosis area was disappeared. But there were remained few hepatocytes lived in these areas. The liver cells mostly are double core cells. Plenty of inflammatory cells infiltrated to the area of necrosis and bleeding. Vacuolar degeneration and apoptotic bodies were remained occurred in some area of the liver tissue.Big area of bleeding and necrosis were occurred more frequently in the liver tissues at 36h post modeling. Lots of vacuolar degenerations were occurred at the edge of necrosis and bleeding area. Plenty of big granules of lipid droplets were accumulated in lived liver cells. Many inflammatory cells infiltrated to the area of bleeding and necrosis apoptotic bodies were remained occurred in some area.The area of bleeding and necrosis were extended and more frequently occurred in all the visual field in the liver tissues at 48h post modeling. Many of new bleeding area were found. Lobule structure of most of necrosis and bleeding area was disappeared. Inflammatory cells infiltrated to the area of bleeding and necrosis. Apoptotic bodies were remained occurred in some area of the liver cells.Most of the liver tissue was occupied by the bleeding and necrosis at 72h post modeling. Many double nucleus cells were occurred in the periphery necrosis area. Inflammatory cells were infiltrated to the area of bleeding and necrosis as the state of 48h post modeling. New bleeding and apoptotic bodies also occurred in the liver tissues.Big area of bleeding and necrosis were also occurred in most of the liver tissues at 120h post modeling. But the new bleeding was markedly decreased. Few of inflammatory cells infiltrated to the area of bleeding and necrosis. Liver cell nucleuses were heavy-stained revealed plenty of liver cells were tended to apoptosis.3Dynamic changes of liver injury related cytokines mRNA expressed by injured mice liver caused by DMN.The RNA were extracted from collected liver tissue samples and reverse transcripted. mRNA of Fas,TNF-α,MMP-2,MMP-3,MMP-9,JNK-1,JNK-2 tested with real-time Quantitative PCR method. The results showed that... |
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