Application of cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-like inhibitory protein to treatment of fatty livers

A fatty liver and liver technology, applied in gene therapy, peptide/protein components, preparations for in vivo experiments, etc., can solve the problem of no Caspase-8 proteolytic enzyme activity, etc., and achieve the effect of inhibiting fatty liver

Inactive Publication Date: 2017-03-15
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein is structurally similar to Caspase-8, but has no proteolytic activity of Caspase-8

Method used

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  • Application of cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-like inhibitory protein to treatment of fatty livers
  • Application of cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-like inhibitory protein to treatment of fatty livers
  • Application of cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-like inhibitory protein to treatment of fatty livers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] [Example 1] Obtaining mouse fatty liver (diet induced obesity, DIO)

[0058] (1) Grouping of experimental animals: 8-week-old, male, cFLIP-flox mice and cFLIP-KO mice were selected and given two special feeds, D12942 high-fat diet (High fat diet, HFD) and D12450B low-fat diet ( Normal chow, NC) feeding, that is, cFLIP-flox NC group, KO NC group, cFLIP-flox HFD group, KO HFD group, a total of 4 groups.

[0059] (2) The model induces the operation process through high-fat feed:

[0060] Using cFLIP-flox and KO mice, the DIO model was established, and phenotype correlation analysis was performed to clarify the role of cFLIP gene on fatty liver. Eight-week-old, male, cFLIP-flox mice and cFLIP-KO mice were selected and fed with two special diets, D12942 high-fat diet (Highfat diet, HFD) and D12450B low-fat diet (Normal chow, NC), respectively, namely There are 4 groups including cFLIP-flox NC group, KO NC group, cFLIP-flox HFD group, and KO HFD group. The fasting body wei...

Embodiment 2

[0061] [Example 2] Determination of mouse body weight

[0062] ①Fasting: fast the mice to be tested at 8:00 am (without water), and start the experimental operation at 2:00 pm.

[0063] ② Weighing: Weigh at 0 week, 4 week, 8 week, 12 week, 16 week, 20 week and 24 week respectively, put a small plastic bucket on the dynamic electronic balance, grab the mouse, put it into the weighing In a small bucket, measure the weight and record the data. Feed amount detection: After the weighing operation is completed, add feed to the mice, and record the amount of feed for the mice on the dynamic electronic balance.

[0064] Weight change results such as figure 1 As shown, cFLIP-flox mice were fed with HFD diet, and their body weight was significantly higher than that of their NC diet group from the 4th week. The body weight of cFLIP-KO mice in the HFD group was significantly higher than that of cFLIP-flox mice in the HFD group starting at week 8, and continued until week 24 (see figu...

Embodiment 3

[0065] [Example 3] Liver Gross Appearance Determination and Pathological Staining

[0066] (1) Terminal liver tissue collection

[0067] 1) After the mice were weighed, they were quickly killed by decapitation. Fix the mouse in a supine position, and moisten the hair on the chest and abdomen of the mouse with distilled water.

[0068] 2) Clamp the skin in the middle of the mouse abdomen with a pair of tweezers, cut the skin along the middle of the abdomen toward the head to the xiphoid process, and cut the skin toward the tail to expose the subcutaneous fascia and muscles layer by layer. organ.

[0069] 3) Quickly find and remove the liver of the mouse, place the removed liver specimen on sterilized gauze, wipe off the residual blood on the surface of the liver, place the liver in a sterile petri dish, and weigh it quickly. Part of the liver tissue after weighing was used to prepare frozen and paraffin sections, and the other part was stored in a -80°C refrigerator for dete...

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Abstract

The invention discloses a function and an application of cFLIP (cellular Fas-associated death domain-like interleukin-1 beta-converting enzyme-like inhibitory protein) genes to fatty livers. The application takes cFLIP liver specific gene knockout mice and cFLIP-flox mice as experiment objects and adopts a diet-induced obesity mouse model, results show that compared with the cFLIP-flox mice, the weight of the cFLIP-knockout mice is obviously higher than that of the cFLIP-flox mice fed by same feed, gross appearances of mouse livers, weight, ratio of liver to weight, lipid composition pathological staining results and the like show that lesion of the fatty livers of the high-fat diet cFLIP-knockout mice is obviously serious, and lipid accumulation is remarkably increased. The cFLIP can serve as selection of drug targets for treating the fatty livers, and accelerants of the cFLIP can be used for preparing the drugs for treating the fatty livers.

Description

technical field [0001] The invention belongs to the field of gene function and application, in particular to a cellular Fas-associated death domain-like interleukin-1β-converting enzyme (FLICE)-like inhibitory protein protein, cFLIP) as a target gene in the preparation of drugs for the prevention, alleviation and / or treatment of fatty liver. Background technique [0002] Fatty liver refers to the lesion with excessive accumulation of fat in liver cells caused by various reasons. It is a relatively common clinical adverse manifestation, which can be induced by various diseases. Mild cases are asymptomatic, and severe cases can be life-threatening. Fatty liver includes alcoholic liver disease and nonalcoholic fatty liver disease, both of which are common chronic liver diseases worldwide. Nonalcoholic fatty liver disease is a metabolic stress disease closely related to insulin resistance and genetic susceptibility. With the continuous improvement of people's living standards...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K49/00A61K38/17A61P1/16
CPCA61K38/1709A61K48/005A61K49/0008
Inventor 李红良王丕晓
Owner WUHAN UNIV
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