The Liver Tissue Gene Expression Profile Of DMN Induced Mouse Liver Injury And The Effect Of Chinese Medicine

Because of the complexity of liver organ, the mechanism of liver injury also complex. Different kind of liver injury showed different mechanisms. So for every kind of liver injury, the mechanism need plenty of research work.We had use DMN injected to the mice to modeling the severe hepatitis. The liver pathological changes are similar to human severe hepatitis. The main liver pathological changes include focal zone of massive bleeding and necrosis closely distributed in liver tissue. Most of liver tissue structure within the focal zone of massive bleeding and necrosis are disappeared.In this research, we analyzed the changes of gene expression profile of DMN induced mouse severe hepatitis model 48h after DMN modeling. The expression significantly changed genes were identified in this model. Then the effects of Chinese herb complex Gan Du Qaing to the significantly changed genes were evaluated. 1The changes of gene expression profile of DMN induced mouse severe hepatitis model20 Kunmin mice, body weight 18±2g, male, bought form centers of experimental animal of Guangzhou University of Chinese Medicine.10 mice were randomly taken as blank control group which were peritoneally injected with saline in dose of 10ml/kg. Another 10 mice were peritoneally injected with DMN in the dose of 15mg/kg for modeling.50 mg liver tissues of every mouse was harvested at 48h after DMN injection and put into lml of Trizol reagent and then settled in-70℃refrigerator. The liver tissues then send to ShangHai KangChen Bio-tech Inc to conduct analysis of gene expression profile.The results showed that 2331 kind of genes were identified to be changed for 2 more times in 44170 kinds of tested genes. There are 1331 genes up-regulated and 1200 genes down-regulated. There are 99 genes changed for 6 more times include 53 genes up-regulation and 46 genes down-regulation. These changed genes involved in 21 signaling pathways. There are 12 signaling pathways may related to liver injury which include PPAR, Fatty acid metabolism, p53 signaling, Drug metabolism, Arachidonic acid metabolism, Glycerolipid metabolism, Metabolism of xenobiotics by cytochrome P450, ECM-receptor interaction, TGF-beta signaling, Cell cycle and MAPK signaling pathways. 2The verification of significant changed expression of genes in the gene expression profile of DMN induced mouse severe hepatitis model24 Kunmin mice, body weight 18±2g, male.12 mice were randomly taken as blank control group which were peritoneally injected with saline in dose of 10ml/kg. Another 12 mice were peritoneally injected with DMN in the dose of 15mg/kg for modeling. All of the mice were sacrificed and liver samples were collected for quantitative PCR testing at the time point of 48h after modeling. The verification experiments concentrated on the 6 more times changed 15 genes which closely related to liver injury signaling pathway. The results showed that all of the verified genes are meet with the results of the gene expression profile.3The effect of Chinese herb complex to the genes closely related to liver injury signaling pathway in DMN induced mouse severe hepatitis model.60 kunming mice were randomly divided into blank control group, model control group, Chinese medicine High (20g/kg.time), middle(10g/kg.time), low dose group(5g/kg.time) and Bifendate control group (150mg/kg.time). There are 10 mice in every group. All of the mice except blank group were injected with DMN in the dose of 15ul/kg for a single dose and start drug administration simultaneously. The drugs were administrated one time every 12 hours. After 48h of drug administration, The mice were sacrificed and serum and liver tissue were collected. Serum was put into-70℃refrigerator. The liver tissue samples were put into 1ml of Trizol reagent for mRNA testing. The results showed that the Chinese herb complex Gan Du Qing can significantly decrease the level of glutamate-pyruvate transaminase in serum which verified the liver protection activity of Gan Du Qing again. The results also showed that the Chinese herb complex Gan Du Qing can significantly affect the mRNA levels of 9 genes in 15 tested genes. The 9 genes are CPT1C,CYP7A1,CDKN1A,THBS1,GSTA1,FMO3,MGLL,TGFB3,HSPB1.These results revealed that the mechanisms of effects of the Chinese herb complex Gan Du Qing to the DMN induced mice severe hepatitis involved in multi-pathways of liver injury. But the Gan Du Qing doesn't affect the mRNA levels of some related genes of liver injury.Conclusion:1. The results of gene expression profile testing showed that there are 2331 genes were identified to be changed for 2 more times in 44170 kinds of tested genes. There are 1331 genes up-regulated and 1200 genes down-regulated. There are 99 genes changed for 6 more times include 53 genes up-regulation and 46 genes down-regulation. These changed genes involved in 21 signaling pathways.15 genes are closely related to liver injury signaling pathway.2. The Chinese herb complex Gan Du Qing can significantly affect the mRNA levels of 9 genes in 15 tested genes. The 9 genes are CPT1C,CYP7A1,CDKN1A,THBS1,GSTA1,FMO3,MGLL,TGFB3,HSPB1. These results revealed that the mechanisms of effects of the Chinese herb complex Gan Du Qing to the DMN induced mice severe hepatitis involved in multi-pathways of liver injury.