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Use of hepatocyte nuclear factor-1alpha in treatment of chronic liver disease

A chronic liver disease, gene therapy technology, applied in genetic engineering and clinical medicine such as disease treatment, molecular biology, cell biology

Active Publication Date: 2012-07-11
上海赛迪夫医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Up-regulation of HNF1α expression has not been studied as an anti-hepatic fibrosis treatment at home and abroad.

Method used

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  • Use of hepatocyte nuclear factor-1alpha in treatment of chronic liver disease
  • Use of hepatocyte nuclear factor-1alpha in treatment of chronic liver disease
  • Use of hepatocyte nuclear factor-1alpha in treatment of chronic liver disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Differential expression of HNF1α protein and mRNA in normal human liver tissue and liver cirrhosis tissue detected by immunohistochemistry and Real-time PCR

[0037] 1 Immunohistochemical method was used to detect the changes of HNF1α protein in liver tissue.

[0038] Human normal liver tissue and liver cirrhosis tissue wax block 4mm serial section, baked in 60℃ oven for 30min and fixed, dewaxed to water, 3%H 2 o 2Place at room temperature for 10 minutes to remove endogenous peroxidase, and microwave citrate buffer for antigen retrieval, add 1:20 normal rabbit serum to block at room temperature for 30 minutes, add HNF1α antibody (1:200) dropwise at 4°C overnight; the next day PBS (0.01 M, pH 7.4) for 3 times, 5 min each time; add secondary antibody and incubate at room temperature for 30 min; after washing with PBS, add SABC (1:100) and incubate at room temperature for 20 min, develop color with DAB, mount with conventional resin, and observe under a light microscope. ...

Embodiment 2

[0057] Inhibitory effect of up-regulated HNF1α expression on liver fibrosis progression in rats.

[0058] 1 Liver fibrosis model preparation:

[0059] DMN injury liver fibrosis model: Male SD rats were randomly divided into 4 groups, 10 in each group. Common food feeding, free drinking water, day and night lighting alternately. Group 1 received intraperitoneal injection of normal saline as the negative control group; Groups 2 to 4 were the liver fibrosis model group, and received intraperitoneal injection of 1% DMN solution at a dose of 10 μg / kg, three times a week for a total of 4 weeks. A rat liver fibrosis model induced by DMN was prepared. BDL liver fibrosis model: 40 male SD rats were divided into 4 groups, 10 in each group. Common food feeding, free drinking water, day and night lighting alternately. The first group is the sham operation group, and the second to fourth groups are the BDL group. After BDL rats were anesthetized by intraperitoneal injection of 10% chl...

Embodiment 3

[0066] Real-time PCR and immunohistochemical methods confirmed: HNF1α significantly inhibited HSC activation and extracellular matrix deposition in vivo, and blocked the EMT process of liver fibrosis.

[0067] 1 Extract total RNA from liver tissue of rats in each group of DMN and BDL liver injury models: 0.5-0.8g liver tissue, add TRizol reagent (1ml / 100mg tissue), crush the tissue until it is homogenized, and place it at room temperature for 5min; add chloroform 0.2 ml / ml TRizol, shake vigorously for 15s, then let stand at room temperature for 3min; Centrifuge at 12000rpm for 10min; discard the supernatant, add absolute ethanol 1ml / ml TRizol, and vortex to mix; centrifuge at 4°C, 7500rpm for 5min, discard ethanol, and dry naturally at room temperature; add 50μl DEPC water to dissolve the RNA; after the RNA is fully dissolved 1 μl each was subjected to 1.5% agarose gel electrophoresis, and the optical density values ​​at 260 nm and 280 nm wavelengths were measured with a UV sp...

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Abstract

The invention relates to use of hepatocyte nuclear factor-1alpha in treatment of liver disease, particularly to use of hepatocyte nuclear factor-1alpha (HNF1alpha) in treatment of liver fibrosis and cirrhosis. The study of the invention shows that regulation of the expression of HNF1alpha gene can effectively generate an induction effect on the hepatocyte nuclear factor, so as to provide a new means of treatment.

Description

technical field [0001] The present invention relates to the fields of molecular biology, cell biology, genetic engineering and clinical medicine such as disease treatment. Specifically, the present invention relates to the use of hepatocyte nuclear factor 1α (HNF1α) to treat human chronic liver disease and liver cirrhosis. The present invention also relates to a method for introducing HNF1α gene and protein into human chronic liver disease, liver cirrhosis liver tissue and cells and a means for increasing the expression of HNF1α in these tissues and cells. Background technique [0002] Chronic liver disease and liver cirrhosis are common chronic diseases in clinical practice. They are difficult to treat, have poor clinical prognosis, occupy huge medical resources, and are major diseases that endanger national health. Hepatic fibrosis is an inevitable stage in the development of various chronic liver diseases to cirrhosis. It is a repair response of the liver to chronic dama...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/17A61K31/7088A61P1/16
CPCA61K48/00A61K38/17A61K31/7088A61P1/16
Inventor 谢渭芬林勇曾欣钱慧施健
Owner 上海赛迪夫医药科技有限公司
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