Application of PQBP1 protein and coding gene thereof in medicine for preventing or treating obesity and non-alcoholic fatty liver disease

Abstract

The invention discloses application of PQBP1 protein and a gene thereof in drugs for preventing or treating obesity and non-alcoholic fatty liver disease. The invention discloses a shRNA (short hairpin ribonucleic acid) molecule, a recombinant adenovirus vector and an application of the shRNA molecule and the recombinant adenovirus vector in a medicine for preventing or treating obesity or non-alcoholic fatty liver disease. The invention proves that the deficiency of PQBP1 in adipose tissues under normal diet conditions can cause the down-regulation of lipid synthesis related enzyme expression, which shows adipogenesis disorder. The deficiency of PQBP1 in adipose tissue under a high fat diet condition resists the obesity process and improves glucose resistance and the fatty degeneration level of non-alcoholic fatty liver. In addition, the further development of obesity can be slowed down by reducing the expression of the PQBP1 in adipose tissues during early obesity, and related metabolic defects caused by obesity are improved. Therefore, related reagents inhibiting the PQBP1 can provide a new treatment strategy for obesity and related metabolic complications.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the application of PQBP1 protein and its encoding gene in drugs for preventing or treating obesity and non-alcoholic fatty liver disease. Background Art

[0002] Adipose tissue is an important energy storage and endocrine organ in mammals, and is involved in the body's glucose and lipid metabolism balance and the development of most tissues and organs. Disorders in human adipose tissue generation, such as obesity caused by excessive fat accumulation, are predisposing factors for a variety of diseases such as type 2 diabetes, cardiovascular disease, non-alcoholic fatty liver disease, and even cancer. At present, obesity and the metabolic-related diseases it causes are showing a rapid development trend internationally, seriously threatening people's health and quality of life. In addition, the current clinical drugs on the market have problems such as poor efficacy and large side effects. Therefore, in-depth exploration of new regulatory factors in the processes of adipose tissue development and homeostasis maintenance is crucial for the target development of obesity-specific drugs.

[0003] Clinical studies have found that patients with Renpenning syndrome caused by mutations in glutamine-binding protein 1 (PQBP1) have a common characteristic of thin stature and a typical lipodystrophy phenotype. Interference with the expression of PQBP1 in nematode intestinal cells and primary white adipocytes of mice can lead to impaired adipogenesis, suggesting that PQBP1 may be involved in the development of adipose tissue and is a potential new target for the treatment of adipose tissue generation disorders.

[0004] The polyglutamine binding protein 1 (PQBP1) gene was first discovered in a human cDNA library in the 1990s. The gene is located on the X chromosome, and its encoded product, the PQBP1 protein, can directly bind to the polyglutamine sequence, the causative source of hereditary neurodegenerative diseases, with extremely high affinity (Iwamoto et al. 2000). Studies have shown that mutations in the PQBP1 gene lead to the occurrence of Renpenning syndrome, a type of mental retardation disease. Most patients are male and often show clinical features such as severe intellectual disability, microcephaly and small testicles (Germanaud et al. 2011). At the same time, it is worth noting that among the pathogenic mutation sites reported so far, about 80% of the sites will cause patients to show thin body characteristics. However, it is still unclear how PQBP1 regulates the adipogenesis process. Summary of the invention

[0005] Purpose of the invention: The technical problem to be solved by the present invention is to provide the use of PQBP1 protein or its inhibitor in drugs for preventing or treating obesity and non-alcoholic fatty liver disease.

[0006] The technical problem that the present invention needs to solve is to provide a shRNA molecule and a recombinant adenovirus vector thereof.

[0007] The final technical problem to be solved by the present invention is to provide the use of the shRNA molecule, the recombinant adenovirus vector or the mouse obtained by the method in a drug for preventing or treating obesity or non-alcoholic fatty liver disease.

[0008] Technical solution: In order to solve the above technical problems, the present invention provides the use of PQBP1 protein or its inhibitor in drugs for preventing or treating obesity and non-alcoholic fatty liver disease, and the genbank accession number of the amino acid sequence of the PQBP1 protein is NP_001239458.2 (mouse source) or NP_001027555.1 (human source).

[0009] The present invention also includes the use of the PQBP1 gene encoding the PQBP1 protein in drugs for preventing or treating obesity and non-alcoholic fatty liver disease. The PQBP1 gene sequence encoding the PQBP1 protein has a genbank Gene ID: 54633 (mouse source) or Gene ID: 10084 (human source).

[0010] Wherein, the application includes down-regulating the expression of PQBP1 protein.

[0011] Wherein, the application includes knocking out the gene encoding PQBP1 protein.

[0012] Among them, the application includes constructing adipose tissue-specific knockout mice of the PQBP1 gene.

[0013] The present invention also includes a shRNA molecule, the nucleotide sequence of the shRNA molecule is shown as SEQ ID NO.1.

[0014] The present invention also includes a recombinant adenovirus vector, which contains the shRNA molecule according to claim 6.

[0015] The present invention also includes a method for constructing a PQBP1 gene knockout mouse, comprising the following steps: injecting the recombinant adenovirus vector into the subcutaneous fat of wild-type mice to obtain the vector.

[0016] The present invention also includes the use of the shRNA molecule, the recombinant adenovirus vector or the mouse obtained by the method in drugs for preventing or treating obesity or non-alcoholic fatty liver disease.

[0017] The present invention utilizes PQBP1 AKOThe study used mice as the research subjects and found that PQBP1 regulates the formation of white adipose tissue by affecting the lipid synthesis process. AKO The weekly food intake and body weight of mice did not change significantly, but their white fat deposition decreased significantly. Transcriptome sequencing analysis of subcutaneous white adipose tissue showed that PQBP1 AKO The expression of lipid synthesis-related enzymes in mice, such as Acly, Acaca, Elovl6, and Dgat2, decreased significantly, which ultimately led to a reduction in the volume of white adipocytes and blocked lipogenesis, but did not damage the metabolic homeostasis of individuals. If the above findings still play a role in the process of diet-induced obesity, it is expected to provide new therapeutic targets and strategies for the treatment of obesity. Under high-fat feeding conditions, we found that PQBP1 AKO The weight gain rate of mice was significantly reduced, and the amount of white fat produced was significantly reduced. At the same time, they did not develop metabolic diseases caused by obesity such as glucose resistance and severe non-alcoholic fatty liver disease, and maintained normal basal metabolic levels. These results suggest that PQBP1 has great application potential in the prevention of obesity. In order to further identify its application value, the present invention synthesized a recombinant adenovirus vector containing shRNA molecules that specifically interfere with the expression of PQBP1. Among them, the specific interference target sequence SEQ ID NO.1 of the recombinant adenovirus vector is: 5-CCAAGAAAGTCAGGAACAA-3. The interfering recombinant adenovirus vector was injected into the subcutaneous white fat of wild-type mice with early obesity by in situ injection, and it was found that the weight gain of mice was significantly slowed down during the subsequent high-fat feeding process, and the degree of glucose resistance and non-alcoholic fatty liver disease was improved. In summary, these results suggest that inhibiting the expression of PQBP1 can provide a new treatment method for obesity and related metabolic complications.

[0018] Beneficial effects: The present invention uses Cre-LoxP technology to construct adipose tissue-specific PQBP1 knockout transgenic mice (PQBP1 AKO The present invention has demonstrated that under normal dietary conditions, the loss of PQBP1 in adipose tissue will lead to down-regulation of the expression of enzymes related to lipid synthesis, which manifests as impaired lipogenesis. Under high-fat dietary conditions, the loss of PQBP1 in adipose tissue will resist the obesity process and improve the level of glucose resistance and fatty degeneration of non-alcoholic fatty liver. In addition, reducing the expression of PQBP1 in adipose tissue during the early stages of obesity can also slow down the further development of obesity and improve related metabolic defects caused by obesity. Therefore, the related reagents that inhibit PQBP1 can provide a new treatment strategy for obesity and related metabolic complications. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 PQBP1 gene adipose tissue-specific knockout mice (PQBP1 AKO A, WB detection of PQBP1 AKO Mouse knockout efficiency. BAT, brown fat; iWAT, inguinal white fat; eWAT, epididymal white fat; testis, testis; liver, liver. B, WB grayscale analysis results. C, adipose tissue generation weight. D, adipose tissue deposition. E, representative adipose tissue pictures. F, three types of adipose tissue weight statistics. G, representative white adipose tissue HE sections. H, iWAT average adipocyte diameter. I, eWAT average adipocyte diameter. *P<0.05, **P<0.01.

[0020] Figure 2 PQBP1 AKO Impaired lipid production in subcutaneous white fat of mice. A, Volcano plot of transcriptome sequencing analysis. B, GO enrichment analysis of differentially upregulated genes. C, GO enrichment analysis of differentially downregulated genes. D, qRT-PCR validation results of differentially upregulated genes. E, qRT-PCR validation results of differentially downregulated genes. *P<0.05, **P<0.01

[0021] Figure 3 PQBP1 AKO Mice have normal basal metabolic levels under standard diet feeding conditions. A, Representative images of liver HE and Oil Red O staining. B, Triglyceride, glucose, high-density lipoprotein and low-density lipoprotein test results. C, GTT experiment. D, ITT experiment. E, Metabolic cage monitoring indicators, 24h oxygen consumption statistics. F, 24h respiratory entropy statistics. H, 24h heat production statistics. Results are expressed as mean ± standard error.

[0022] Figure 4 PQBP1 AKO Mice resist the obesity phenotype induced by a high-fat diet. A, Weekly weight curve. B, Representative pictures of mice. C, Fat deposition. D, Representative pictures of adipose tissue and liver. E, Tissue weight statistics. F, Tissue deposition statistics. G, Representative white adipose tissue HE-stained sections. H, Average diameter of iWAT adipocytes. I, Average diameter of eWAT adipocytes. *P<0.05, **P<0.01.

[0023] Figure 5 PQBP1 AKOMice resist obesity-related metabolic diseases induced by a high-fat diet. A, Representative images of liver HE and Oil Red O staining. B, GTT experiment. C, ITT experiment. D, Metabolic cage monitoring indicators, 24h oxygen consumption curve. E, Metabolic cage monitoring indicators, 24h carbon dioxide emission curve. F, Metabolic cage monitoring indicators, 24h carbon dioxide emission, respiratory entropy and heat production statistics. *P<0.05, **P<0.01.

[0024] Figure 6 Results of inhibiting the development of obesity in initially obese mice by recombinant adenovirus that interferes with PQBP1 expression. A, virus injection strategy. B, weekly body weight curve. C, fat deposition. D, representative images of inguinal white fat HE staining, liver HE staining, and Oil Red O staining. E, GTT experiment. F, ITT experiment. *P<0.05, **P<0.01. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is described in detail below in conjunction with the accompanying drawings and embodiments.

[0026] 1. Experimental Animals

[0027] The PQBP1-flox strain mice used in the experiment were owned by our laboratory (constructed by the Model Animal Institute of Nanjing University, and the relevant articles have been published: Shen, Y., Zhang, ZC, Cheng, S., et al. (2021). PQBP1 promotes translationalelongation and regulates hippocampal mGluR-LTD by suppressing eEF2phosphorylation. Mol. Cell 81, 1425-1438.e10.). The PQBP1 gene of this mouse contains the same flox sequence on both sides of exon 3 and exon 6; the AdipoQ-Cre strain mice were donated by Li Chaojun's research group at Nanjing University (purchased from the Jackson Laboratory, https: / / www.jax.org / jax-mice-and-services). Based on the knockout principle of the Cre-Loxp system, the PQBP1-flox strain mice and the AdipoQ-Cre strain mice can be hybridized to construct adipose tissue-specific knockout PQBP1 mice (denoted as PQBP1 AKO The specific construction strategy is to select female PQBP1-flox homozygous mice (genotype PQBP1 fl / fl ) were crossed with male AdipoQ-Cre+ mice, and the genotype of the obtained offspring was detected, among which the genotype was AdipoQ-Cre+; PQBP1 fl / Y PQBP1AKO Mice, genotype AdipoQ-Cre-;PQBP1 fl / Y The mice were used as control mice. In addition, the wild-type mice used in the experiment were purchased from Jicui Pharmaceutical Company. All strains of mice were of C57 / BL6J background and were kept in the Experimental Animal Center of the Jiangbei Campus of Southeast University. The temperature was kept at a constant temperature of 22°C, the light cycle was 12h light / 12h dark, and the light start time was set at 7 am. All animal husbandry and experiments were carried out in accordance with the relevant regulations of Southeast University.

[0028] 2. PQBP1 AKO Reduced white fat production in mice

[0029] To determine the PQBP1 AKO To determine whether the mouse was successfully constructed, we selected 3-month-old PQBP1 AKO Male mice were killed by cervical dislocation, and their subcutaneous inguinal white fat (iWAT), epididymal gonadal white fat (eWAT), scapular brown fat (BAT), testicular tissue and liver tissue were weighed, and then the proteins of each tissue were extracted for Western Blot detection. According to the test results, PQBP1 AKO The expression levels of different PQBP1 proteins in mice were significantly reduced, while they were normal in non-adipose tissues, indicating that the tissue-specific knockout mice were successfully constructed ( Figure 1 AB). Further tissue generation analysis results showed that under standard diet feeding conditions, PQBP1 AKO The total fat production of mice decreased significantly, and the fat deposition index was also significantly lower than that of the control group mice ( Figure 1 CD). Specifically, white fat production was significantly reduced, while brown fat production did not change significantly ( Figure 1 To further determine the anatomical changes, we took PQBP1 AKO Fresh iWAT and eWAT tissues of mice were paraffin-sectioned in the order of fixation-dehydration-transparent-wax-embedding-sectioning, and the tissue structure was observed by HE staining. AKO The average diameter of white adipocytes in mice was significantly lower than that in control mice ( Figure 1 GI), indicating that the ability of adipose tissue to store lipids is reduced, ultimately leading to a decrease in white fat production.

[0030] 2.PQBP1 AKO Impaired lipogenesis in white adipose tissue of mice

[0031] To further determine whether PQBP1 AKOTo investigate the molecular expression changes in white adipose tissue of mice, we selected 3-month-old control mice (sample size n = 2) and PQBP1 AKO The iWAT tissue of mice (sample size n = 2) was used to extract RNA and perform transcriptome sequencing analysis. The process mainly includes three parts: RNA sequencing library construction, sequencing on the machine, and statistical analysis of sequencing data. First, VAHTS was used to analyze the RNA. TM Stranded mRNA-Seq Library Prep Kit for The sample RNA was processed with a kit (NR602, Novozyme) to construct an individual transcriptome library. Subsequently, bridge PCR amplification was performed on a cBot instrument to generate clusters, and sequencing was performed using the Illumina platform. All sequencing processes were completed at Shanghai Jingneng Biotechnology Co., Ltd.

[0032] After the original sequencing data was provided by Jingneng, we first used an internal script edited by Perl language to perform quality control on all the original sequencing data, including the removal of the adapter data in the original data, the removal of the data with the measured RNA ploy-N content exceeding 10%, and the removal of the data with poor quality, etc., and finally obtained pure sequencing data, namely clean reads, and its Q20, Q30 and GC content were counted. Next, using the mouse reference genome mm_10 (GCA_000001635.2) as a template, the clean reads were aligned and spliced ​​using Hisat 2 (v2.0.9) software, and then the counts of different genes were calculated using FeatureCounts (v2.1.1) software, and the data were used for subsequent differential analysis (Trapnell et al. 2010). The differential gene expression analysis was calculated using the edgeR package, where the screening criteria for differential genes were the absolute value of the differential change fold ≥ 2 and PValue < 0.05. For the differentially expressed genes obtained by analysis, functional enrichment analysis can be performed using the GO (https: / / david.ncifcrf.gov / ) and KEGG (http: / / www.genome.jp / kegg / ) websites to preliminarily determine the differentially expressed genes.

[0033] The results of bioinformatics analysis showed that a total of 515 differentially expressed genes were screened, including 388 differentially up-regulated genes and 127 differentially down-regulated genes ( Figure 2 A). GO enrichment analysis was performed on these two types of genes, and the results showed that the differentially upregulated genes were mainly involved in biological processes such as immune response activation and regulation ( Figure 2B), the differentially down-regulated genes are mainly involved in metabolic-related biological processes such as fatty acid metabolism ( Figure 2 C). Next, we used qRT-PCR to verify the accuracy and reliability of the sequencing analysis results. -△△Ct The expression of each gene was detected by qRT-PCR. The specific operation was carried out according to the instructions of the fluorescent quantitative dye TB Green (Cat. No.: TKR-RR820A, Manufacturer: TaKaRa).

[0034] The specific PCR system is shown below.

[0035]

[0036] The experimental instrument was a CFX96 Real-Time PCR instrument, and the program was as follows: 95°C, 30 s; 95°C, 5 s, 60°C, 30 s, 39 cycles; melting curve. Each gene was tested three times, and the relative expression of the gene was measured using 2 -△△Ct The primer sequences were shown in Table 1.

[0037] Table 1. qRT-PCR primers

[0038]

[0039]

[0040] The gene expression trend in this experimental result is basically consistent with the sequencing analysis results ( Figure 2 DE).

[0041] 3. PQBP1 AKO Reduced white fat production in mice does not lead to an imbalance in individual metabolic homeostasis

[0042] When the production of fat tissue in the body is disordered, it is very easy to cause an imbalance in individual metabolic homeostasis, such as ectopic fat deposition or insulin resistance. Therefore, we then identified PQBP1 from different aspects. AKO Basic metabolic status of mice. First, the liver tissue, the main organ for ectopic fat deposition, was tested. AKO Fresh liver tissues of male mice were paraffin-embedded and frozen-embedded. HE staining of paraffin-embedded sections revealed that PQBP1 AKO The liver tissue of mice was relatively compact and did not show pathological features such as fibrosis or vacuolization. At the same time, the results of Oil Red O staining of frozen liver sections showed that PQBP1 AKOThe liver tissue of the mice did not show abnormal lipid accumulation and was in a healthy state ( Figure 3 A). The levels of triglycerides, glucose, high-density lipoprotein and low-density lipoprotein in the blood are PQBP1 AKO Normal levels in mice ( Figure 3 B).

[0043] Next, we investigated PQBP1 AKO The glucose tolerance of mice was tested. The specific implementation plan was to fast the mice for 16 hours, weigh them and use a blood glucose meter to measure the glucose content in the tail tip blood, which was recorded as 0-min blood glucose. Then, the mice were injected with intraperitoneal glucose solution at a dosage of 2g / kg (glucose amount / mouse body weight), and the blood glucose levels of the mice were measured at 30min, 60min, 90min and 120min after the injection. After the measurement was completed, the mice were allowed to resume their diet. The experimental test results showed that PQBP1 AKO The mice did not show glucose resistance, and their blood glucose curves showed no significant difference from those of the control mice ( Figure 3 C). At the same time, we also investigated PQBP1 AKO The insulin sensitivity of mice was tested. After fasting for 4 hours, the mice were weighed and their blood glucose levels were measured at 0 min. Then, 0.5 IU / kg (insulin amount / mouse body weight) of insulin solution was injected into the mice intraperitoneally, and the blood glucose levels of the mice were measured at 30 min, 60 min, 90 min and 120 min after the injection. The recorded results showed that PQBP1 AKO 90 minutes and 120 minutes after insulin injection, the blood glucose levels of mice were significantly lower than those of the control group, and the blood glucose recovery rate was significantly reduced, suggesting that PQBP1 AKO The mice had normal insulin sensitivity and increased insulin action time ( Figure 3 D) In ​​addition, we used the PhenoMaster multifunctional metabolic analysis system to analyze the expression of PQBP1 AKO The basal metabolism of mice was monitored and it was found that PQBP1 AKO The oxygen consumption, respiratory entropy (carbon dioxide production / oxygen consumption) and heat production of mice during the day and night were similar to those of control mice ( Figure 3 EG). It can be seen that under standard feeding conditions, PQBP1 AKO The reduced white fat production in mice did not cause metabolic disorders in the individuals.

[0044] 4. PQBP1 AKO Obesity induced by a high-fat diet can be suppressed in mice

[0045] Due to PQBP1 AKOMice will develop a phenotype of white adipocyte production disorder under normal diet feeding conditions. Therefore, in order to verify and further explore the underlying reasons why these mice can resist obesity-like conditions induced by a high-fat diet, 8-week-old PQBP1 AKO The mice and their littermates were subjected to an 8-week high-fat obesity modeling experiment. According to the weekly body weight records, PQBP1 AKO The weight of mice was significantly lower than that of mice in the control group from the second week of high-fat feeding, and the weight difference widened further with the increase of feeding time ( Figure 4 AB). At the 8th week of high-fat diet, tissue sampling and analysis of the two groups of mice revealed that PQBP1 AKO The amount of fat tissue deposits in mice was significantly lower than that in control mice ( Figure 4 C), among which brown fat production did not change significantly, while white fat production decreased significantly ( Figure 4 DF). HE staining results of paraffin sections also further showed that PQBP1 AKO The white fat cells of mice were significantly smaller than those of control mice ( Figure 4 GI). These results suggest that PQBP1 AKO The mice inhibited the development of obesity mainly by inhibiting the hypertrophy of white fat cells.

[0046] 5. PQBP1 AKO Obesity-induced metabolic disturbances alleviated in mice

[0047] To further verify that PQBP1 AKO We further investigated whether mice could also improve metabolic disorders caused by obesity. AKO The liver health of mice and their littermates (i.e. PQBP1 non-knockout mice) showed that after 8 weeks of high-fat diet, the livers of the littermates showed obvious vacuolation, while the livers of PQBP1 AKO This phenomenon was not observed in mice, and their slice structures still showed a compact and dense phenotype. At the same time, Oil Red O staining results showed that a large number of lipid droplets accumulated in the liver of control mice, while only a small number of small lipid droplets were found in the knockout mice ( Figure 5 A). The results of glucose tolerance test (GTT) and insulin tolerance test (ITT) also showed that PQBP1 AKO The mice had normal blood sugar regulation and did not develop glucose intolerance ( Figure 5 BC). In addition, the results of the basic metabolic detection system analysis found that PQBP1 AKO Mice had an increased oxygen consumption rate at night ( Figure 5 D), and the carbon dioxide excretion rate and respiratory entropy (RER) were significantly higher than those of the control mice ( Figure 5EG), but its heat production did not change ( Figure 5 H). In conclusion, these results suggest that PQBP1 AKO In the process of resisting obesity, the mice also alleviated the pathological metabolic disorders induced by obesity.

[0048] 6. Interference with PQBP1 expression in early obese mice can inhibit the development of obesity and related metabolic diseases

[0049] In order to effectively transfer the above results to the development of clinical drugs for obesity treatment, we selected wild-type male mice (C57 / BL6J background) and injected Pqbp1 interference recombinant adenovirus and control virus into the inguinal white fat at 6 weeks of age by multi-point in situ injection, that is, 3 injection points were selected on each side of the fat, and the injection volume on each side was 1E+11vg ( Figure 6 A). The interference vector used was pAAV-U6-shRNA / spgRNA v2.0-CMV-EGFP-WPRE (Heyuan Biotechnology (Shanghai) Co., Ltd., catalog number: H12663), the target sequence of the shRNA of the Pqbp1 gene was 5'-CCAAGAAAGTCAGGAACAA-3', and the target sequence of the control group was 5'-TTCTCCGAACGTGTCACGT-3'. The synthesis of this part of the virus was completed by Heyuan Biotechnology (Shanghai) Co., Ltd.

[0050] Subsequently, at 8 weeks of age, the diet of the two groups of mice was changed to high-fat feeding, and an 8-week obesity modeling experiment was conducted. The weekly weight records showed that the weight of mice injected with the Pqbp1 interference virus was significantly lower than that of the control group in the third week of high-fat feeding, and the difference further increased with the increase in feeding time ( Figure 6 B). After 8 weeks of feeding, the fat deposits in mice injected with Pqbp1 interference virus were significantly lower than those in the control group ( Figure 6 C), specifically, the production of inguinal white adipose tissue was significantly reduced, corresponding to a significant decrease in the volume of white adipocytes ( Figure 6 D). At the same time, from the metabolic level, the livers of mice injected with the Pqbp1 interference virus still showed a compact phenotype, without vacuolization and abnormal lipid accumulation ( Figure 6 D). The results of the glucose tolerance test (GTT) and insulin tolerance test (ITT) also further showed that the blood sugar regulation ability of mice injected with the Pqbp1 interference virus was normal and there was no glucose intolerance ( Figure 6 EF). Therefore, the experimental results of this part suggest that inhibiting the expression of PQBP1 in adipose tissue in the early stage of obesity can effectively prevent the occurrence of obesity and maintain normal metabolic homeostasis of individuals.

Claims

1. Use of PQBP1 protein or its inhibitor in a drug for preventing or treating obesity and non-alcoholic fatty liver disease, characterized in that: The genbank accession number of the amino acid sequence of the PQBP1 protein is NP_001239458.2 or NP_001027555.

1.

2. Encoding PQBP1 protein PQBP1 The use of genes in drugs for preventing or treating obesity and non-alcoholic fatty liver disease is characterized in that: The coding PQBP1 protein PQBP1 The genbank Gene ID of the gene sequence is 54633 or GeneID: 10084.

3. The use according to claim 1, characterized in that: The application includes down-regulating the expression of PQBP1 protein.

4. The use according to claim 2, characterized in that: The application includes knocking out the gene encoding PQBP1 protein.

5. The use according to claim 2, characterized in that: The application includes constructing adipose tissue-specific knockout mice of the PQBP1 gene.

6. A shRNA molecule, characterized in that The nucleotide sequence of the shRNA molecule is shown in SEQ ID NO.

1.

7. A recombinant adenovirus vector, characterized in that: The recombinant adenovirus vector contains the shRNA molecule according to claim 6.

8. A PQBP1 A method for constructing a knockout mouse, characterized in that: The method comprises the following steps: injecting the recombinant adenovirus vector according to claim 7 into the subcutaneous fat of wild-type mice to obtain the vector.

9. Use of the shRNA molecule according to claim 6, the recombinant adenovirus vector according to claim 7 or the mouse obtained by the method according to claim 8 in a drug for preventing or treating obesity or non-alcoholic fatty liver disease.

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