Studies On Acute And Chronic Liver Injury In Mice By Molecular Biology Techniques

Study on several molecular indexes in diagnosis of alcoholic liver disease in miceThe mice model of alcoholic liver injury should be set up to evaluate several genes could be adapted for candidate indexes for diagnosing alcoholic liver disease. The mice were treated with alcohol. After 6 weeks and 12 weeks, compared with the normal mice, the activity of alanine aminotransferase (ALT) in serum in alcohol-treated mice was increased;while the concentrations of total proteins (TP), albumin(ALB) were reduced markedly. In pathological sections of liver tissue, destruction of the structure of hepatocytes, steatosis, and lymphocytes soakage were found in mice treated with alcohol. The mice model of alcoholic liver injury was established successfully. Then, RNA was extracted from liver tissues of normal mice and alcohol-treated mice, and was reverse-transcripted into cDNA. Six genes such as CD14, LBP, MCP-1, ICAM-1, IL-10, OPN were amplified by PCR in liver tissues of normal mice and alcohol-treated mice respectively. These genes of alcohol-treated mice were expressed at levels higher than those of normal mice. Thus, the results suggest these genes could be the candidate indexes in diagnosis of alcoholic liver disease by molecular biology method.Construction of the subtracted cDNA library of differentially expressed genes in mice liver of CCl₄-induced acute injury and normal liverHuman genomic technology make the differential expression of function genes get more and more popular. To find differentially expressed genes becomes most hot in the molecular biology research. Here, we make the differentially expressed cDNA subtracted library in mice liver treated with CCl₄ compared with normal mice liver. First, mice model of CCl₄-induced acute liver injury should be set up. Mice were intraperitoneally (ip.) administered 40% CCl₄: peanut oil. After sixteen hours, compared with normal mice, the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) in serum were increased in model mice, while the concentrations of total proteins(TP), albumin(ALB) in serum were reduced markedly. In pathological sections, the blur of hepatic lobule, the hyperemia, the ballooning degeneration of hepatocyte were found in the liver tissues of the mice treated with CCl₄. The mice model of CCl₄-induced acute liver injury was set up successfully. To find the differentially expressed genes in mice liver treated with CCl₄ compared with normal mice liver and make the differentially expressed cDNA subtracted library, suppression subtractive hybridization (SSH) was applied. The differentially expressed fragments were obtained through twice hybridizations and twice PCR, then T/A cloning was accomplished to construct the cDNA library. Last, sixty positive cloneswere randomly picked out and identified by PCR, Fifty-five contain positive insert fragments which range from 200 to 750 bp. A subtracted cDNA library of differentially expressed genes in mice liver of CCU-induced acute injury and normal liver is successfully constructed with SSH and T/A cloning techniques. This experiment has prepared for the cloning and identification of the full-length genes which were differentially expressed. Furthermore, it also helps to exploit gene remedy and diagnosis gene chip.