The applications of gene Grp75 in the preparation of drugs for the treatment of liver fibrosis
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A technology of liver fibrosis and liver fibrosis, applied in the field of molecular biology, can solve the problem of lack of constitutive expression
Inactive Publication Date: 2013-07-17
FUDAN UNIV +1
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However, Grp75 is not constitutively expressed in liver tissue (Li Hua, Yang Zengjie, Xia Beili, Zuo Ji. Expression characteristics of glucose-regulated protein 75. [J]. Fudan Journal (Medical Science Edition), 2001, 28(5): 382 ), how to transfer the gene of GRP75 to up-regula
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Embodiment 1
[0030] Example 1 Cell experiment
[0031] 1 Establishment and identification of cellular oxidative damage model
[0032] 1.1 Cells and main reagents:
[0033]7702 cells were purchased from cells of the Chinese Academy of Sciences, thiazolyldiphenyl-tetrazolium bromide (MTT) was purchased from amresco, dimethyl sulfoxide (DMSO) was purchased from Merck, and high-glucose medium (DMEM) was purchased from invitrogen. h 2 o 2 purchased from Shanghai Sangong.
[0034] 1.2 Experimental steps:
[0035] Take 7702 cells in the logarithmic growth phase, make a single cell suspension, and use 5×10 4 Cells / ml were inoculated in 96-well culture plates, with 6 replicate wells in each group, two groups of cells in the normal group and cells in the GRP75 up-regulation group, 200 μl of culture medium at 37°C, 5% CO 2 After culturing in the incubator for 12 hours, add 400uM H 2 o 2 Continue to culture for 2 hours, then carry out MTT detection, that is, discard the cell culture supernatan...
Embodiment 2
[0105] Embodiment 2 animal experiments
[0106] 1. Establishment of Rat CCL 4 Induced liver fibrosis model
[0107] 1.1 Animals and reagents
[0108] CCL 4 ; glycerol; normal saline.
[0109] SPF grade Wistar rats were used in the experiment.
[0110] 1.2 Grouping
[0111] The SPF grade Wistar rats used in the experiment were raised in an animal room with a room temperature of 22°C and a relative humidity of 55%, with a 12-hour circadian rhythm, free access to food and water, and the experiment began after 1 week of adaptation to the environment. The rats were randomly divided into 12 control groups, 12 model groups, 12 GRP75 transgenic groups, and 12 GRP75 upregulation + injury groups. All index tests were performed after a 12-hour fast. All rats were controlled at a body weight of 100-150g.
[0112] 1.3 Experimental steps
[0113] The normal rats (model group) fed for 5 days were given intraperitoneal injection of 10% CCL diluted with glycerol 4 , injected at a dos...
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Abstract
The invention belongs to the field of molecular biology, relating to the applications of Grp75 in the preparation of drugs for the treatment of oxidative damages of liver cells and liver fibrosis by using gene transfection techniques. According to the invention, based on that Grp75 is an important chaperone in cytoplasm and mitochondria, wherein the 1-23 loci of the N-terminal thereof is a mitochondrial transmembrane signal, can help a protein to be correctly folded, assist the transport of the protein, and has the characteristics such as removal of ROS and antiapoptosis, experiments of cytoprotective effects of Grp75 on H2O2-induced oxidative stress in cells, and protective effects on CCL4-induced hepatic fibrosis as a pathological basis in animal models are carried out. When an injury occurred in the liver cells, the GRP75 gene is transfected in the cells, and the expressions of Grp75 protein are upregulated respectively at a cells level and a whole level; and results show that the effect of the treatment of liver fibrosis can be achieved through the effects of anti-oxidative damage and inhibition of hepatocyte apoptosis of the Grp75 protein. The Grp75 gene of the present invention can be used for preparing drugs for the treatment of liver fibrosis.
Description
Technical field: [0001] The invention belongs to the field of molecular biology, and relates to a new use of Grp75 gene, in particular to the use of Grp75 in the preparation of medicines for treating oxidative damage to liver cells and liver fibrosis by applying gene transfection technology. Background technique: [0002] Liver fibrosis is the wound healing response of the liver to liver injury caused by various reasons, which is manifested by the proliferation and deposition of connective tissue in the liver. Modern research has shown that most of the common multiple acute and chronic liver diseases such as viral hepatitis, fatty liver, alcoholic liver and so on are manifested as liver fibrosis in the early stage. If not detected and effectively suppressed, it may further develop into liver cirrhosis or even Liver cancer directly endangers the patient's life. According to research, normal liver cell damage is regarded as the initial signal to activate liver fibrosis. With ...
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