Application of liposome nanoparticles in targeted intervention of liver CD36 expression in treatment of lean fatty liver

Abstract

The invention relates to application of liposome nanoparticles in targeted intervention of liver CD36 expression in treatment of lean fatty liver. The invention relates to treatment of lean fatty liver, and particularly discloses application of Si CD36 mRNA liposome nanoparticles in targeted intervention of liver CD36 treatment. Research finds that expression increase of CD36 is related to lean fatty liver, and finds that after target tail intravenous injection of the Si CD36 mRNA liposome nanoparticles, fatty degeneration and inflammatory response of the liver can be inhibited. Compared with a wild type mouse fed with high fat diet, the expression increase of CD36 in the mouse fed with high fat diet in the daytime can be inhibited by the Si CD36 mRNA nano-liposome particles, so that the fatty degeneration and inflammatory response of the liver are inhibited. The invention provides a new therapeutic target for diagnosing the thin fatty liver, and provides a new method for treating the thin fatty liver.

Description

Technical Field

[0001] The present invention relates to the treatment of lean non-alcoholic fatty liver disease, and in particular to the use of SiC36 mRNA liposome nanoparticles targeting liver CD36 to treat lean non-alcoholic fatty liver disease. Background Art

[0002] Unlike traditional non-alcoholic fatty liver disease (NAFLD), which mainly affects overweight and obese people, lean NAFLD can occur in people with normal weight, making the diagnosis and management of the disease more complex and challenging. In recent years, the prevalence of lean NAFLD has gradually increased, which may be related to the increase in metabolic syndrome, sedentary lifestyles, and dietary changes. Similar to NAFLD in overweight people, it may lead to insulin resistance, type 2 diabetes, and increased risk of cardiovascular disease. In addition, lean NAFLD may also progress to non-alcoholic steatohepatitis (NASH), liver fibrosis, and even cirrhosis, which carries serious long-term health risks.

[0003] Si mRNA liposome nanoparticles are an emerging biomedical technology that can effectively deliver messenger RNA (mRNA) to target cells by encapsulating it in liposomes, thereby guiding the cells to produce specific proteins. Compared with traditional gene delivery methods, Si mRNA liposome nanoparticles have higher safety and efficiency, while avoiding the immune response and pathogenic risks that may be caused by viral vectors. The advantage of this technology lies in its high plasticity, which can adjust the composition and structure of liposomes according to different treatment needs, thereby optimizing drug release and cellular uptake.

[0004] CD36 is a transmembrane glycoprotein widely distributed on the surface of various cell types and belongs to the scavenger receptor family. It plays an important role in physiological processes such as immune regulation, lipid metabolism and cell signal transduction. Recent studies have shown that CD36 is closely related to the occurrence and development of various metabolic diseases, such as diabetes and non-alcoholic fatty liver disease. Therefore, CD36 has not only attracted attention in basic medical research, but also become a potential therapeutic target, providing new ideas for the intervention and treatment of related diseases. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the purpose of the present invention is to provide a new treatment method for lean fatty liver.

[0006] In order to achieve the purpose of the present invention, the present invention firstly synthesized Si CD36mRNA liposome nanoparticles.

[0007] The inventors found that a high-fat daytime diet can induce the occurrence of lean fatty liver in mice compared with the livers of mice fed with ordinary feed.

[0008] CD36(Cluster of Differentiation 36)GenBank NO.**NM_001001506.3

[0009] The present invention provides a means of targeted intervention of CD36 expression in the liver, wherein the synthesized SiCD36 mRNA liposomes are injected into mice from the tail vein, targeted to the liver of mice with lean fatty liver, and the pathological manifestations of lean fatty liver are suppressed.

[0010] Tail vein injection of SiCD36 mRNA liposomes inhibited the development of lean fatty liver in mice compared with wild-type mouse livers

[0011] Therefore, the Si CD36 mRNA liposome targeted intervention of CD36 expression in the liver reduces fatty degeneration of the liver of the organism, and can also be used to reduce the inflammatory response of the organism. The beneficial effects of the present invention are:

[0012] The present invention found that the increased expression of liver CD36 is correlated with lean fatty liver, and found that SiCD36 mRNA liposome targeted intervention inhibited liver fatty degeneration and inflammatory response. Studies have found that restricting mice to a high-fat diet during the day can cause lean fatty liver in mice, and targeted intervention of CD36 expression in the liver of mice by Si CD36mRNA lipid nanoparticles can reduce fatty degeneration and inflammatory response in the liver of mice. The present invention provides a new target for studying new drugs for treating lean fatty liver. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 These are pathological sections of the liver of mice fed a normal diet (ad libitum diet NA, daytime diet ND, nighttime diet NN) and mice fed a high-fat diet (ad libitum diet HA, daytime diet HD, nighttime diet HN), mainly to observe hepatic fatty degeneration and hepatic lipid droplet accumulation.

[0014] Figure 2 Levels of liver injury markers in the serum of mice fed a normal diet (ad libitum diet NA, daytime diet ND, nighttime diet NN) and mice fed a high-fat diet (ad libitum diet HA, daytime diet HD, nighttime diet HN); N = 8 (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001).

[0015] Figure 3Quantification of mRNA of inflammatory factors in the liver of mice fed with a normal diet (ad libitum diet NA, daytime diet ND, nighttime diet NN) and mice fed with a high-fat diet (ad libitum diet HA, daytime diet HD, nighttime diet HN).

[0016] Figure 4 The expression of CD36 in the liver of mice fed with a normal diet (ad libitum diet NA, daytime diet ND, nighttime diet NN) and mice fed with a high-fat diet (ad libitum diet HA, daytime diet HD, nighttime diet HN) (*, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001).

[0017] Figure 5 Model flow chart for injection of Si CD36 mRNA liposome nanoparticles.

[0018] Figure 6 Pathological sections and serum liver injury marker levels of mice injected with Si CD36 mRNA liposome nanoparticles, N = 8 (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001). Specific implementation methods

[0019] The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention.

[0020] Example 1

[0021] 1. Materials and Methods

[0022] 1. Animal model and treatment: Six-week-old C57 / BL6J male mice were selected and fed with normal diet (NCD) and high-fat diet (HFD, 60% kal, D12492, Research Diets Inc) for ad libitum feeding during the day and night for 16 weeks, and the mice fed ad libitum served as the control group. Mice were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd.

[0023] 2. HE staining: After paraffin embedding, the tissue was sliced, dewaxed and hydrated, dripped with hematoxylin for 5 minutes, rinsed with running water, then dripped with eosin for 2 minutes, rinsed with running water, then dehydrated and sealed, and imaged with an automatic slice scanner for observation and analysis.

[0024] 3. Oil Red O staining: frozen tissue sections, fixed with 4% paraformaldehyde, stained with Oil Red O solution for 10-20 min, counterstained with hematoxylin for 1-2 min, observed and photographed under a microscope, and analyzed the lipid droplet infiltration in the liver.

[0025] 4. Real-time fluorescence quantitative PCR: Tissue RNA was extracted with Trizol (TAKARA, Japan) and then reverse transcribed and subjected to real-time fluorescence quantitative PCR detection. 2. Results

[0026] The inventors found that mice fed a high-fat diet (HD) had increased lipid accumulation in the liver ( Figure 1 ). Liver damage markers were significantly increased in the serum of mice fed a high-fat diet (HD) ( Figure 2 At the same time, we found that the expression of inflammatory factors in mice increased through real-time fluorescence quantitative PCR ( Figure 3 ). Through transcriptomics detection and real-time fluorescence PCR, we found that CD36 expression in mice was significantly increased ( Figure 4 ).

[0027] Example 2

[0028] 1. Materials and Methods

[0029] 1. The solvent in the phospholipid solution was evaporated in a nitrogen stream to obtain a thin layer of phospholipids. An aqueous dispersion of Si-RNA complexes in phosphate buffered saline (PBS) at pH 7.4 was added to the film and stirred continuously at 60°C for 30 minutes. The solution was extruded through a polycarbonate membrane with a pore size of 200 nm for 10 cycles to obtain liposomes of uniform size, which were then freeze-dried. Example 3

[0030] 1. Materials and Methods

[0031] 1. Animal model and treatment: C57 / BL6J mice that had been fed a high-fat diet for 12 weeks were selected and the obtained liposome nanoparticles were injected into the mice through the tail vein ( Figure 5 ) and then fed with a high-fat diet (HFD) for 2 weeks before sampling ( Figure 5 ).

[0032] 2. Liver HE staining, real-time fluorescence quantitative PCR: The methods are the same as those described in Example 1.

[0033] 2. Results

[0034] Histological examination results showed that the liver fatty degeneration of mice injected with nanoliposome particles was significantly reduced, and the infiltration of inflammatory cells was also reduced. The levels of liver injury markers in serum were reduced ( Figure 6 ).

[0035] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. Application of SiCD36 mRNA nanoliposomes in the treatment of lean non-alcoholic fatty liver disease.

2. An application for treating lean non-alcoholic fatty liver disease, characterized in that: The application targets liver CD36.

3. The application according to claim 3, characterized in that: The use can reduce fatty degeneration of the liver of an organism.

4. The use according to claim 3, characterized in that: The use can reduce the inflammatory response of the liver of the organism.

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