CN111920951B - Application of ZNRF3/RNF43 in preparation of drugs for regulating energy metabolism - Google Patents
Application of ZNRF3/RNF43 in preparation of drugs for regulating energy metabolism Download PDFInfo
- Publication number
- CN111920951B CN111920951B CN202010692053.XA CN202010692053A CN111920951B CN 111920951 B CN111920951 B CN 111920951B CN 202010692053 A CN202010692053 A CN 202010692053A CN 111920951 B CN111920951 B CN 111920951B
- Authority
- CN
- China
- Prior art keywords
- znrf3
- rnf43
- mice
- liver
- energy metabolism
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
Abstract
The invention relates to an application of ZNRF3/RNF43 in preparing a drug for regulating energy metabolism. The ZNRF3/RNF43 double knockout is proved to reduce the blood sugar content and the serum insulin level, increase the sensitivity of an organism to glucose and regulate the expression of gluconeogenesis related genes; meanwhile, the liver lipid deposition is reduced, the expression of lipid synthesis and inflammation related genes is reduced, and the results show that ZNRF3/RNF43 double knockout can improve liver insulin resistance by influencing gluconeogenesis, lipid synthesis and other ways, so that a theoretical basis is provided for preparing the energy metabolism regulating drug.
Description
Technical Field
The invention belongs to the field of type 2 diabetes mellitus medicines, and particularly relates to application of ZNRF3/RNF43 in preparation of a medicine for regulating energy metabolism.
Background
The ring finger proteins ZNRF3 (zinc ring finger protein 3) and RNF43 (ring finger protein 43) are two homologous single transmembrane E3 ubiquitin ligases, both belong to the giant human (Goliath) protein family, serve as conserved genes of various organisms, and are stably expressed in various vertebrates. ZNRF3 is located on human chromosome 22, RNF43 is located on chromosome 17, and the two structures are similar and have certain homology and are composed of four parts, namely a Signal Peptide (SP), an extracellular domain (PA region), a transmembrane domain (transmembrane domain) and a functional domain which plays a ubiquitination role in cells, namely a RING domain, and the ZNRF3 and the RNF43 are functional homologues. Ubiquitination refers to the specific modification of substrate protein by ubiquitin molecules (ubiquitin) to mediate the degradation and elimination of the substrate protein. The ZNRF3/RNF43 protein is coded by downstream genes of a Wnt signal pathway, can be combined with a DEP (Dishevelled, egl-10and Plecktrin domain) of Dishevelled protein (DVL) to recognize that Wnt receptor Frizzled (FZD) and low-density lipoprotein receptor-related protein 6 (LRP 6) can ubiquitinate the protein and degrade the protein through an endocytosis-lysosome pathway, so that the protein is cleared from a cell membrane, thereby inhibiting the Wnt signal pathway and being an important Wnt pathway antagonist protein; can also combine with a cysteine-rich Fu1 domain (furin-like domain) of R vertebra protein (Rspondin, rspo), and simultaneously combine with LGR4/5/6 to form a ZNRF3/RNF43-Rspo-LGR4/5/6 complex, thereby losing the function of negatively regulating the Wnt signaling pathway and helping to maintain the activation state of the Wnt signaling pathway. The Wnt signaling pathway is involved in many embryonic processes and tissue homeostasis, and is closely related to energy metabolism. At present, ZNRF3/RNF43 is mainly researched in the aspects of tumorigenesis, tissue development and the like, and no research in the aspects of energy metabolism and the like exists.
Disclosure of Invention
The invention aims to solve the technical problem of providing an application of ZNRF3/RNF43 in preparing a medicament for regulating energy metabolism, and discussing the effect of ZNRF3/RNF43 on energy metabolism.
The invention provides an application of ZNRF3/RNF43 in preparing a medicament for regulating energy metabolism.
Preferably, ZNRF3/RNF43 is used as a target gene and is matched with pharmaceutically acceptable auxiliary materials or auxiliary components to prepare a preparation for use.
Preferably, the preparation is selected from one of tablets, powders, granules, capsules, oral liquids and sustained release agents.
The invention adopts Cre-loxP recombinant enzyme system to carry out liver specificity double knockout on ZNRF3/RNF43, carries out high fat diet induction on ZNRF3/RNF43 double knockout mice and control group mice to establish an obesity model, utilizes a sugar tolerance test and serum insulin detection to carry out related molecular biology analysis, and observes the influence on the sugar metabolism of the mice; observing body fat content and blood fat level, detecting the level of relevant factors of lipid metabolism, and discussing the influence of ZNRF3/RNF43 on lipid metabolism. Meanwhile, adenovirus is used for carrying out specific ZNRF3 overexpression on the liver, and the function of ZNRF3/RNF43 in body energy metabolism is further verified.
Advantageous effects
The ZNRF3/RNF43 double knockout is proved to reduce the blood sugar content and the serum insulin level, increase the sensitivity of an organism to glucose and regulate the expression of gluconeogenesis related genes; meanwhile, the liver lipid deposition is reduced, the expression of lipid synthesis and inflammation related genes is reduced, and the results show that ZNRF3/RNF43 double knockout can improve liver insulin resistance by influencing gluconeogenesis, lipid synthesis and other ways, so that a theoretical basis is provided for preparing the energy metabolism regulating drug.
Drawings
FIG. 1a is a glucose tolerance test of a control group mouse and a ZNRF3& RNF43 gene double knockout mouse; b is serum insulin levels in two groups of mice; c is the expression of two groups of mouse gluconeogenic genes.
FIG. 2a shows the result of HE staining of liver of control mice and ZNRF3& RNF43 gene double knockout mice (200 ×); b is the hepatic triglyceride content of the two groups of mice; and c is the total cholesterol content of the livers of the two groups of mice.
FIG. 3 shows liver lipid synthesis (a) and inflammation-related factor (b) expression in control mice and ZNRF3& RNF43 gene double knockout mice.
FIG. 4a is the body weight of control group mice and ZNRF3 overexpression group mice; b is the hepatic triglyceride content of the two groups of mice; c is the expression of genes related to metabolism in the liver.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Example 1
1. Animal experiments
ZNRF3& RNF43 liver-specific double knockout mutant male mice (C57 BL/6) are bred in the animal breeding center of Shanghai university of transportation and are fed at SVF level. Single cages were housed and HFD diet started at 8 weeks of age, with normal diet using standard commercial mouse diet (21% protein, 4.5% fat, 4% cellulose, caloric 1.404 kcal/g), high fat diet purchased from Research Diets (20% carbohydrate, 20% protein, 60% fat) without restriction of drinking and food intake. The ambient temperature was set at a constant temperature of 22 ℃, 12-hour light cycle (6. The breeding mode is that 8-week-old male mice and 8-week-old female mice are mated in a cage according to the ratio of 1.
Genetically deficient obese mice were 8 week old, male, healthy SPF grade b6.Bks (D) Leprdb/JNju (db/db) mice purchased from the university of Nanjing institute of model animals. Mice were housed in the animal house of the institute of endocrine research, shanghai university of transportation medical school and were fed with normal feed (65% carbohydrate, 22% protein, 5% fat, 8% cellulose, caloric value 3.3 kcal/g) without restriction of drinking water and food intake. The ambient temperature was constant at 22 ℃,12 hours light cycle (8.
2. Results of the experiment
(1) The liver specificity double knockout ZNRF3& RNF43 can reduce the blood sugar of mice and improve the sugar tolerance.
The results of the glucose tolerance test performed on mice and the evaluation of the sensitivity of the mice to systemic glucose show that the blood glucose of ZNRF3/RNF43 double knockout mice at each time point is obviously lower than that of control group mice, and the blood glucose differences at 0 minute, 15 minute, 30 minute, 60 minute, 90 minute and 120 minute have statistical significance (P < 0.05) (figure 1 a), which indicates that the sensitivity of ZNRF3/RNF43 double knockout mice to glucose is increased. Further detecting the serum insulin level of the mice, the result shows that the serum insulin of the ZNRF3/RNF43 double knockout mice is obviously lower than that of the control group mice (figure 1 b), which indicates that the ZNRF3/RNF43 double knockout mice can improve the glucose tolerance and hyperinsulinemia caused by insulin resistance of the mice.
In order to evaluate the gluconeogenesis capability of the mice, real time PCR detection is carried out on key genes of gluconeogenesis in livers, and the results show that the expression of genes such as PEPCK, G6PASE, HNF1a, HNF4a and the like related to gluconeogenesis in the livers of ZNRF3/RNF43 double-knockout mice is obviously reduced (figure 1 c), which indicates that the gluconeogenesis in the livers can be inhibited by ZNRF3/RNF43 double-knockout.
(2) Liver-specific double knockout of ZNRF3& RNF43 improved liver lipid deposition in mice.
Liver HE staining results showed that lipid droplets were reduced in hepatocytes from ZNRF3/RNF43 double knockout mice (fig. 2 a), indicating improved liver lipid deposition. Correspondingly, when the lipid content in the liver of the mouse is detected, the liver triglyceride content of the ZNRF3/RNF43 double knockout mouse is obviously reduced (figure 2 b) and the total cholesterol content of the liver is obviously reduced (figure 2 c) compared with the control group of mice, and the difference has statistical significance.
(3) Liver-specific double knockout of ZNRF3& RNF43 reduces liver lipid synthesis and inflammation-related factor expression in mice.
In order to discuss the mechanism of ZNRF3/RNF43 double knockout mice for improving liver lipid deposition, realtime PCR was further used to detect the expression of genes related to lipid metabolism in the liver. Analysis results show that ZNRF3 and RNF43 gene double knockout mice livers have significant down-regulation of ZNRF3 gene, RNF43 gene and genes closely related to lipid synthesis, such as FAS, SCD1, SREBP1c and the like (FIG. 3 a), which indicates that intrahepatic lipid synthesis is reduced. When the expression of factors related to liver tissue inflammation is further detected, the expression of inflammatory factors such as MRC2, CCL8, TNFa and the like in the liver of a ZNRF3& RNF43 gene double-knockout mouse is obviously reduced (figure 3 b), which indicates that the ZNRF3& RNF43 gene double-knockout mouse can possibly improve intrahepatic inflammation.
(4) The effect of liver-specific overexpression on the metabolism of obese mice.
Based on the affinity of liver to adenovirus, ad-ZNRF3 packaged by recombinant adenovirus and its control virus are subjected to overexpression of ZNRF3 in liver by means of tail vein injection with db/db mouse as target, and the weight change of two groups of mice within 6 weeks after virus injection is observed and recorded. It was found that the body weight of mice injected with Ad-ZNRF3 adenovirus was significantly increased (FIG. 4 a) and the hepatic triglyceride levels significantly increased (FIG. 4 b) compared to mice injected with control virus.
Meanwhile, after the Realtime PCR analysis of the genes related to lipid metabolism in the liver, the liver of the mouse injected with the Ad-ZNRF3 adenovirus is found to be up-regulated in the expression of genes such as PEPCK, G6PASE, HNF4a and the like related to gluconeogenesis and obviously up-regulated in the expression of genes such as FAS, SCD1, SREBP1c, PGC1a and the like related to lipid synthesis (figure 4 c), and the role of ZNRF3/RNF43 in the energy metabolism of organisms is further verified.
3. Conclusion of the experiment
The results show that ZNRF3/RNF43 double knockout can reduce blood sugar content and serum insulin level, increase the sensitivity of the organism to glucose and down-regulate gluconeogenesis-related gene expression; meanwhile, liver lipid deposition is reduced, expression of relevant genes of lipid synthesis and inflammation is reduced, and the results show that ZNRF3/RNF43 double knockout can improve liver insulin resistance by influencing gluconeogenesis, lipid synthesis and other ways, and further provide theoretical basis for preparing medicines for regulating energy metabolism.
Claims (3)
1. Use of a ZNRF3/RNF43 double knockout agent for the manufacture of a medicament for modulating energy metabolism by inhibiting gluconeogenesis and down regulating lipid synthesis.
2. Use according to claim 1, characterized in that: ZNRF3/RNF43 is used as a target gene and is matched with pharmaceutically acceptable auxiliary materials or auxiliary components to prepare a preparation for use.
3. Use according to claim 2, characterized in that: the preparation is selected from one of tablets, powder, granules, capsules, oral liquid and sustained release agents.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010692053.XA CN111920951B (en) | 2020-07-17 | 2020-07-17 | Application of ZNRF3/RNF43 in preparation of drugs for regulating energy metabolism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010692053.XA CN111920951B (en) | 2020-07-17 | 2020-07-17 | Application of ZNRF3/RNF43 in preparation of drugs for regulating energy metabolism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111920951A CN111920951A (en) | 2020-11-13 |
| CN111920951B true CN111920951B (en) | 2022-12-06 |
Family
ID=73313694
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010692053.XA Active CN111920951B (en) | 2020-07-17 | 2020-07-17 | Application of ZNRF3/RNF43 in preparation of drugs for regulating energy metabolism |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111920951B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104080471A (en) * | 2011-10-14 | 2014-10-01 | 诺华股份有限公司 | Antibodies and methods for Wnt pathway-related diseases |
-
2020
- 2020-07-17 CN CN202010692053.XA patent/CN111920951B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104080471A (en) * | 2011-10-14 | 2014-10-01 | 诺华股份有限公司 | Antibodies and methods for Wnt pathway-related diseases |
Non-Patent Citations (3)
| Title |
|---|
| Genomic landscape of colitis-associated cancer indicates the impact of chronic inflammation and its stratification by mutations in the Wnt signaling;Masashi Fujita,et al;《Oncotarget》;20181231;第9卷(第1期);第969-981页 * |
| ZNRF3对类风湿关节炎滑膜增殖和炎症的作用及其分子机制;梁菁菁;《中国优秀博硕士学位论文全文数据库(博士)医药卫生科技辑》;20191015(第10期);第E065-19页 * |
| 环指蛋白4通过TAK1抑制MAPK信号通路减轻心梗后炎症反应;吉家钗等;《心脏杂志》;20200225;第32卷(第1期);第14-19页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111920951A (en) | 2020-11-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Chen et al. | Four decades after the discovery of regenerating islet-derived (Reg) proteins: current understanding and challenges | |
| Ingersoll et al. | The role and pathophysiological relevance of membrane transporter PepT1 in intestinal inflammation and inflammatory bowel disease | |
| Leturque et al. | GLUT2 mutations, translocation, and receptor function in diet sugar managing | |
| Sodhi et al. | Toll-like receptor-4 inhibits enterocyte proliferation via impaired β-catenin signaling in necrotizing enterocolitis | |
| Lees et al. | Genetics of inflammatory bowel disease: implications for disease pathogenesis and natural history | |
| CN102483419B (en) | Utilize the method for apparent metabolic transformation agent, multidimensional intracellular molecules or environment affecting agent diagnosis dysbolism | |
| Pilz et al. | Visfatin/pre-B-cell colony-enhancing factor: a protein with various suggested functions | |
| Jiang et al. | Parabiotic heterogenetic pairing of Abcc6−/−/Rag1−/− mice and their wild-type counterparts halts ectopic mineralization in a murine model of pseudoxanthoma elasticum | |
| Makki et al. | Beneficial metabolic effects of rapamycin are associated with enhanced regulatory cells in diet-induced obese mice | |
| CN107624116B (en) | Peptide having anticancer activity, and pharmaceutical composition and dietary supplement composition for preventing and treating cancer containing the same as active ingredient | |
| Powis et al. | Molecular pharmacology and antitumor activity of palmarumycin-based inhibitors of thioredoxin reductase | |
| van Der Zande et al. | The helminth glycoprotein omega‐1 improves metabolic homeostasis in obese mice through type 2 immunity‐independent inhibition of food intake | |
| CN103648531A (en) | Method for prevention or treatment of metabolic syndrome | |
| Yiew et al. | The mitochondrial pyruvate carrier at the crossroads of intermediary metabolism | |
| Tschurtschenthaler et al. | The selective autophagy receptor optineurin in crohn’s disease | |
| CN107446033A (en) | The treatment of development-related disorders | |
| Wu et al. | Taurine inhibits kupffer cells activation induced by lipopolysaccharide in alcoholic liver damaged rats | |
| Bai et al. | Leptin promotes glycolytic metabolism to induce dendritic cells activation via STAT3-HK2 pathway | |
| Wang et al. | Gomisin D alleviates liver fibrosis through targeting PDGFRβ in hepatic stellate cells | |
| CN111920951B (en) | Application of ZNRF3/RNF43 in preparation of drugs for regulating energy metabolism | |
| Giorgioni et al. | Advances in the development of nonpeptide small molecules targeting ghrelin receptor | |
| Behr et al. | Mild nephrogenic diabetes insipidus caused by Foxa1 deficiency | |
| Kim et al. | Zinc transporter 8 haploinsufficiency protects against beta cell dysfunction in type 1 diabetes by increasing mitochondrial respiration | |
| CA2597708A1 (en) | Metabolic-based methods for modulating gene expression | |
| Wang et al. | Proteomic analysis of lysine acetylation reveals that metabolic enzymes and heat shock proteins may be potential targets for DSS-induced mice colitis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |