Medicine and treatment method for treating fatty liver

A technology of fatty liver and medicine, applied in the field of medicine for diseases

Inactive Publication Date: 2018-10-23
成军
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether it is true that HCBP6 plays a negative regulatory role in the synthesis of TC and TG, or there are other regulatory mechanisms upstream of it, remains to be further studied

Method used

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  • Medicine and treatment method for treating fatty liver
  • Medicine and treatment method for treating fatty liver
  • Medicine and treatment method for treating fatty liver

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: HCBP6 regulates the synthesis and accumulation of TC and TG at the cellular level

[0059] 1. Verify the effect of HCBP6 overexpression and silencing

[0060] The pHCBP6 was transiently transfected into the cells cultured in a six-well plate, and the total protein was extracted after 48 hours. The band at the position of 21kDa was detected by the primary antibody against HCBP6. Compared with the control group, the expression of HCBP6 in the experimental group was significantly increased. . The same experimental method verified the interference effect of si-HCBP6 chemically synthesized by Shanghai Jima Company. Western blot showed that compared with the control group, it can interfere with more than 60% of the endogenous HCBP6 protein (see figure 1 a, 1b).

[0061] 2. The effect of overexpression / silencing HCBP6 on intracellular TC / TG

[0062] After transient transfection of pHCBP6 or si-HCBP6 in L02 and HepG2 cell lines, the content of TC / TG in each group ...

Embodiment 2

[0068] Example 2: Construction of HCBP6 knockout mouse model

[0069] 1. The human gene HCBP6 is the gene 4833415N24Rik (NM_026126.4) in the mouse, which is located on the mouse X chromosome;

[0070] 2. TALEN design and construction, using gene 4833415N24Rik exon 3 as the target of TALEN genome editing;

[0071] 3. Pronuclear injection of specific mRNA (TALEN) in mouse fertilized eggs;

[0072] 4. Obtain F0 generation mice and breed F1 generation: raising of transplant recipient mother mice and birth of F0; mating of chimeric mice F0 and wild mice in cages to obtain germline genetic F1 generation heterozygous mice;

[0073] 5. Obtain homozygous HCBP6 gene knockout mice: F1 generation mice are self-crossed to obtain homozygous HCBP6 gene knockout mice, and the tail genotype identification confirms germline inheritance:

[0074] (1) Genomic DNA was extracted from the tail of HCBP6 knockout mice, HCBP6 was amplified by PCR, the product was verified by gel electrophoresis, and ...

Embodiment 3

[0079] Embodiment 3: Construction of fatty liver mouse model

[0080] 1. Grouping of fatty liver model animals: 10 6-8 week C57BL / 6J male mice and 10 6-8 week HCBP6 knockout male mice (C57BL / 6J strain) were selected and randomly divided into the following 4 groups:

[0081]

[0082] Note: WT, wild-type mice; KO, gene knockout mice; Chow, normal diet-fed; HFD, high-fat diet-fed.

[0083] 2. Construction of fatty liver model:

[0084] (1) Control group mice (Chow): fed with a normal diet (conventional mouse feed from Huafukang Company);

[0085] (2) Fatty liver model group (HFD): fed with a high-fat diet (Whitby Technology Development (Beijing) Co., Ltd.);

[0086] The weight of the mice was weighed every week, and the body weight changes were observed; the food intake of the mice was detected at the fifth week, and the fasting blood glucose of the mice was detected at the 12th week.

[0087] 3. Specimen collection and analysis: 12 weeks after modeling, blood was collected...

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Abstract

The invention relates to a screening method for a medicine for targeting HCBP6 (HCV core-binding protein 6) to treat or prevent simple fatty liver, fatty liver hepatitis (NASH), diabetes, multiple sclerosis (MS) and cardiovascular and cerebral atherosclerosis, and a medicine monomer and a medicinal composition screened by using the method. The HCBP6 can sense the changes of TC and TG levels in hepatocytes, correspondingly change with the changes and regulate the TC and TG levels in the cells in a concussion manner in order to maintain the stability of the internal environment of intracellularTC and TG. An HCBP6 knockout mouse and zebra fish animal model is established. Ginsenoside Rh2 obtained through screening can significantly up-regulate the expression of the HCBP6 gene and inhibit thesynthesis of TC and TG, and has significant therapeutic effects on fatty liver, diabetes, cardiocerebral and vascular diseases, atherosclerosis and other diseases.

Description

technical field [0001] The present invention belongs to the field of pharmacy, and in particular, the present invention relates to a method for targeting HCBP6 gene screening of drugs for treating or preventing diseases such as fatty liver, and the drugs obtained by screening through the above method. Background technique [0002] Fatty liver refers to the pathological changes caused by excessive accumulation of fat in liver cells due to various reasons. According to statistics, 80% of liver cancer is caused by viral hepatitis, and fatty liver is recognized as a common cause of hidden cirrhosis, becoming the second largest liver disease after viral hepatitis. In recent years, the prevalence of fatty liver in my country has been increasing and showing a younger trend. According to statistics, the incidence of fatty liver in China is about 18%, and it is even higher in developed cities. [0003] Fatty liver is not only an independent disease, but also causes many comorbiditie...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/704A61K49/00A61P1/16A61P3/10A61P37/02A61P25/00A61P3/06A61P9/10A01K67/02C12N15/90C12Q1/02
CPCA61K31/704A61P1/16A61P3/06A61P3/10A61P9/10A01K67/02A61K31/575A61K31/58A61K45/00A61K49/00A61P25/00A61P37/02C12N15/113C12N15/873C12N15/90C12Q1/02
Inventor 成军
Owner 成军
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