| Liver injury is the essential pathological changes in almost all the lver diseases.China is a country with quantities of Hepatitis B virus carriers and Hepatitis B patients.There are 100 million hepatitis B virus carriers in China.So research in liver injury is very important to China.Our previous research had established the mouse liver injury model caused by DMN injection and the model had been used to evaluate the liver protection effect of Chinese herb.But the liver injury mechanism of this mouse model had not been identified.In this research,we had tried to elucidate the basic mechanism of the liver injury mouse model caused by DMN.We took the emphesize on observation of the expression of TNF-αand IL-1βin serum and liver tissue.The effect of a Chinese complex to the expression of TNF-αand IL-1βin serum and liver tissue were also evaluated.DMN was peritoneally injected to the mouse in a dose of 15mg/kg to cause liver injury.The serum and liver tissue samples were collected in the time points of 6h,12h,24h,36h,48h,72h,96h,120h post DMN jnjection.The TNF-αand IL-1βserum level were tested with ELISA kit.TNF-α,IL-1βmRNA expression levels were tested with SYBR quantitative PCR assay.TNF-α,IL-1βprotein expression in the liver tissue were evaluated with immunohistochemistry assay.The results showed that TNF-αwas markedly elevated in serum at 24h post DMN injection.The tendency was kept at the time points of 24,36,48h.However,the TNF-αlevel is descended at 72h and a little ascend at 120h post DMN injection while both the TNF-αlevels of the 2 time points are higher than the level of 0h.The serum level of IL-1βwas elevated start from 12h post DMN injection.The tendency was kept at the time points of 12,24,36,48h.However,the IL-1βlevel is descended at 72h and a little ascend at 120h post DMN injection while both the IL-1βlevels of the 2 time points are higher than the level of 0h.The results of TNF-α,IL-1βmRNA quantitative PCR in liver tissue showed that TNF-αmRNA level was elevated at 6h and reached peak at time point of 48h post DMN injection.The high level was kept between time points of 6h and 48h.The TNF-αmRNA level was descended at 72h and a little ascend at 120h post DMN injection which are similar to the TNF-αlevel in serum.6h post DMN injection,IL-1βmRNA in liver tissue start to elevate significantly.The elevation was kept between time points of 6h and 48h and reached peak at time point of 48h post DMN injection.IL-1βmRNA level descended at 72h and a little ascend at 120h post DMN injection.The gross apperance of the model liver showed that the hepatic congestion was severe and severe with the time going and additionally,decreased liver volume and edema was accompanied.Live tissue immunohistochemistry results showed that TNF-αand IL-1βwere expressed both intracellular and extracellular.Both the TNF-αand IL-1βpositive signals are appeared from 6h post DMN injection,positive signals exacerbated with time going.The liver protection mechanism of a Chinese herb complex GanDuQing was explored.The results showed that the TNF-αand IL-1βin serum and liver tissue were significantly down-regulated by GanDuQing herb complex.Both the results of liver tissue mRNA quantitative PCR assay and immunohistochemistry assay were proved this conclusion.The results of these experiments revealed that the liver protection mechanism of GanDuQing herb complex may be down-regulation of the secretion of inflammatory factor.DMN induced liver injury is a kind of chemical liver injury.Our research hadn't been involved in characteristics of chemical liver injury.So this area should our research orientation in the future. |
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