Application of mPGES-2 as target spot in development, screening or preparation of medicine for promoting liver regeneration

Abstract

The invention discloses an application of mPGES-2 as a target spot in developing, screening or preparing a medicine for promoting liver regeneration. The invention proposes that mPGES-2 is a drug target for improving the liver regeneration capacity after partial liver resection for the first time. Experiments show that mPGES-2 liver specific knockdown or knockout can help mice to recover liver weight more quickly after partial liver resection, the liver weight ratio of the mice is increased, and the mPGES-2 shows a promoting effect on liver regeneration. In the process of promoting liver regeneration by mPGES-2 liver specific knockdown or knockout, the levels of glutamic oxalacetic transaminase and glutamic-pyruvic transaminase in serum are not changed, namely, the liver function is not influenced, meanwhile, expression of related genes participating in cell cycle and DNA replication can be remarkably up-regulated, proliferation of liver cells is promoted, and then the volume and the function of the liver are increased or maintained; the compound can be used as a target for promoting liver regeneration, and provides an important theoretical basis for research and development of drugs in the field of clinical liver regeneration.

Description

Technical Field

[0001] The invention relates to the application of mPGES-2 (microsomal prostaglandin Esynthase-2) as a target in the development, screening or preparation of drugs for promoting liver regeneration, and belongs to the technical field of biomedicine. Background Art

[0002] Liver disease has become an extremely serious public health problem in my country, affecting more than 30% of the population, including acute liver disease, chronic liver disease, liver cancer, etc. There are many types of liver diseases, and each of them may cause liver damage to varying degrees, seriously threatening human health.

[0003] Liver regeneration is an important physiological response of the liver when it is damaged. Under physiological conditions, old hepatocytes in the liver die naturally, while new hepatocytes continue to be generated, forming a dynamic balance between old and new cells, which is essential for liver health. Under pathological conditions, such as viral infection, chemical-induced damage or surgical resection, the number of hepatocytes will decrease significantly, which will affect the normal function of the liver. This sharp decrease in number poses a threat to the structure and function of the liver. Therefore, the organism must initiate the corresponding liver regeneration and repair mechanism to meet this challenge. Specifically, specific molecular pathways are activated in the body to trigger the replication mechanism of hepatocytes. At this time, dormant hepatocytes are activated and begin to proliferate. At the same time, the remaining liver tissue will also expand to meet the body's metabolic needs. Ultimately, this series of liver regeneration processes not only promotes the proliferation of hepatocytes and ensures the maintenance of liver function, but also provides important self-repair capabilities for the declining liver. Therefore, in-depth research on the mechanism of liver regeneration is of great significance for the treatment of damage caused by liver disease, provides a new direction for finding new drugs that can promote liver regeneration, and opens up new ideas for the treatment of liver diseases. Summary of the invention

[0004] The main purpose of the present invention is to provide an application of mPGES-2 as a target in the development, screening or preparation of drugs for promoting liver regeneration, so as to overcome the deficiencies in the prior art.

[0005] In order to achieve the above-mentioned invention object, the technical solution adopted by the present invention includes:

[0006] The embodiments of the present invention provide the use of mPGES-2 as a target in the development, screening or preparation of drugs for promoting liver regeneration.

[0007] The embodiments of the present invention also provide the use of mPGES-2 or an inhibitor of its encoding gene in the preparation of a drug for promoting liver regeneration.

[0008] The embodiment of the present invention further provides a pharmaceutical composition for promoting liver regeneration, which comprises: an inhibitor of mPGES-2 or its encoding gene, and a pharmaceutically acceptable carrier.

[0009] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention reports for the first time that mPGES-2 is a drug target for promoting liver regeneration, which plays an important role after 70% partial hepatectomy. The experimental results show that by performing liver-specific knockdown or knockout of mPGES-2 in mice, the recovery of their liver weight can be significantly accelerated, the liver weight ratio of mice can be improved, and the liver regeneration process can be promoted. It is worth noting that during this process, the levels of aspartate transaminase (AST) and alanine aminotransferase (ALT) in serum did not change, indicating that the intervention of mPGES-2 will not have a negative impact on liver function; at the same time, the intervention significantly upregulated the expression of genes related to the cell cycle and DNA replication, further promoted the proliferation of hepatocytes, thereby enhancing or maintaining the volume and function of the liver; these findings provide strong support for mPGES-2 as a potential target for promoting liver regeneration, and lay an important foundation for the future research and development of drugs in the field of liver regeneration. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0011] Figure 1a This is a graph showing the expression of mPGES-2 in mice in a control group (negative control, NC) and a mPGES-2 knockdown (knockdown, KD) group in a typical embodiment of the present invention;

[0012] Figure 1b-Figure 1c This is a graph showing the liver weight ratio results of mice in the NC group and the mPGES-2KD group at 48h and 96h after 70% hepatectomy in a typical embodiment of the present invention;

[0013] Figure 2a This is a graph showing the expression of mPGES-2 in mice in the NC group and the mPGES-2 knockout (KO) group in a typical embodiment of the present invention;

[0014] Figure 2bThis is a graph showing the liver weight ratio of mice in the NC group and the mPGES-2KO group 48 hours after 70% hepatectomy in a typical embodiment of the present invention;

[0015] Figure 3a-Figure 3b This is a graph showing the results of detecting liver function biochemical indices, serum AST and ALT in mice in the NC group and mPGES-2KD group 48h and 96h after 70% liver resection in a typical embodiment of the present invention;

[0016] Figure 3c This is a graph showing the results of detecting liver function biochemical indicators, serum AST and ALT in mice in the NC group and the mPGES-2KO group 48 hours after 70% liver resection in a typical embodiment of the present invention;

[0017] Figure 4 This is a diagram showing the results of Ki-67 immunohistochemical staining in the regenerated liver of mice in the NC group and the mPGES-2KD group at 48h and 96h after 70% hepatectomy in a typical embodiment of the present invention;

[0018] Figure 5 This is a graph showing the results of PCNA immunohistochemical staining in the regenerated liver of mice in the NC group and the mPGES-2KO group 48 hours after 70% hepatectomy in a typical embodiment of the present invention;

[0019] Figure 6a This is a heat map analysis result of RNA-seq differential gene in regenerating liver tissue of mice in NC group and mPGES-2KO group in a typical embodiment of the present invention;

[0020] Figure 6b This is a diagram showing the results of performing KEGG (Kyoto Encyclopedia of Genes and Genomes) database analysis on RNA-seq data of regenerated liver tissues of mice in the NC group and the mPGES-2KO group in a typical embodiment of the present invention;

[0021] Figure 6c This is a diagram showing the results of analyzing the cell cycle pathway in RNA-seq data of regenerating liver tissues of NC group and mPGES-2KO group mice in a typical embodiment of the present invention;

[0022] Figure 6d This is a diagram showing the results of analyzing the DNA replication pathway in RNA-seq data of regenerating liver tissue of NC group and mPGES-2KO group mice in a typical embodiment of the present invention. DETAILED DESCRIPTION

[0023] In view of the defects of the prior art, the inventor of this case has proposed the technical solution of the present invention after long-term research and extensive practice. In order to facilitate the understanding of the present application, the present application will be described in more detail as follows. However, it should be understood that the present invention can be implemented in many different forms and is not limited to the embodiments or examples described herein. On the contrary, the purpose of providing these embodiments or examples is to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0024] Specifically, as one aspect of the technical solution of the present invention, it involves the use of mPGES-2 as a target in the development, screening or preparation of drugs for promoting liver regeneration.

[0025] In some preferred embodiments, the liver regeneration refers to the liver regeneration phenomenon after 70% of the liver is removed.

[0026] In some preferred embodiments, when the drug acts on a mouse model, it can at least inhibit the expression of mPGES-2 in the mouse model, wherein the mouse model is a mouse that has undergone 70% partial liver resection.

[0027] Furthermore, when the drug acts on a mouse model, it can at least increase the regenerated liver weight ratio.

[0028] Furthermore, when the drug acts on a mouse model, it can at least promote the expression of proliferation antigen Ki-67 in regenerating liver cells.

[0029] Furthermore, when the drug acts on a mouse model, it can at least promote the expression of PCNA, a DNA replication marker, in regenerating liver cells.

[0030] Furthermore, when the drug acts on a mouse model, it can at least promote the expression of cell cycle genes in regenerating liver cells.

[0031] Furthermore, when the drug acts on a mouse model, it can at least promote the expression of DNA replication genes in regenerating liver cells.

[0032] Furthermore, when the drug acts on a mouse model, it can at least promote the expression of cell proliferation proteins and DNA replication proteins in regenerating liver cells.

[0033] As another aspect of the technical solution of the present invention, it involves the use of an inhibitor of mPGES-2 or its encoding gene in the preparation of a drug for promoting liver regeneration.

[0034] In some preferred embodiments, the inhibitor is selected from interfering molecules that specifically interfere with the expression of the gene encoding mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its encoding gene.

[0035] As another aspect of the technical solution of the present invention, it also relates to a pharmaceutical composition for promoting liver regeneration, which comprises: an inhibitor of mPGES-2 or its encoding gene, and a pharmaceutically acceptable carrier.

[0036] In some preferred embodiments, the inhibitor is selected from interfering molecules that specifically interfere with the expression of the gene encoding mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its encoding gene.

[0037] The present invention is further described by the following examples: The present invention can be better understood according to the following examples. However, it is easy for those skilled in the art to understand that the specific material ratios, process conditions and results described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0038] Unless otherwise specified, the various raw materials, reaction equipment, testing equipment and testing methods used in the following examples are all well known in the art.

[0039] Example 1: Effect of mPGES-2 knockdown on liver regeneration after 70% liver resection in mice

[0040] (1) Before the experiment, 24 8-week-old male C57BL / 6 mice were randomly divided into a control group and an experimental group according to their body weight. The former was injected with a control virus, while the latter was injected with an mPGES-2 liver-specific knockdown adeno-associated virus, thereby constructing a control group and an mPGES-2 liver-specific knockdown mouse model.

[0041] (2) After 2 weeks of observation, the mice were anesthetized with isoflurane and underwent 70% hepatectomy.

[0042] (3) After disinfecting the mouse abdomen with alcohol, cut the skin and muscles 1-2 cm below the xiphoid process to expose the liver, then press the middle lobe with a cotton swab and cut off the falciform ligament on the middle lobe. During this process, the left lobe of the liver needs to be ligated with a thread and cut off slightly above the ligation site.

[0043] (4) Ligate the middle lobe approximately 2-3 mm below the bifurcation of the middle lobe. Then cut off the middle lobe of the liver, add 2-3 drops of saline, and finally suture the muscle and skin.

[0044] (5) Twelve mice (including 6 mice in the NC group and 6 mice in the mPGES-2KD group) were randomly selected and killed after 48 h and 96 h. The liver weight and body weight of the mice were weighed to calculate the liver regeneration index and compare the ratio of the two.

[0045] (6) Western Blot was used to detect the expression of mPGES-2.

[0046] a. RIPA lysis buffer (brand: Bio-Time) was added to mPGES-2NC and mPGES-2KD tissues respectively (100 mM PMSF was added to each mL of lysis buffer at a ratio of 1:100 before use). After lysis on ice for 30 min, the supernatant was collected by centrifugation at 12,000 rpm for 15 min.

[0047] b. Use BCA protein concentration detection kit (brand: Thermo fisher) for quantification, add 5× loading buffer to the supernatant at a ratio of 1:4, vortex, and heat in a metal bath at 100°C for 10 min.

[0048] c. Total protein was separated by 10% SDS-PAGE gel, separation conditions: 150V, 90min, wet transfer to nitrocellulose membrane, wet transfer conditions: 80V, 90min. Blocked with PBST solution containing 3% bovine serum albumin V for 1h, added rabbit anti-mPGES-2 antibody overnight, washed with TBST 3 times, 5min / time; incubated with secondary antibody at room temperature for 1h; washed with TBST 3 times, 5min / time, and detected by ECL chemiluminescence signal. The expression results of mPGES-2 are shown in Figure 1a shown.

[0049] Data analysis: SPSS16.0 software was used to analyze the experimental data. The t-test was used for comparison between two groups, and one-way ANOVA was used for comparison between multiple groups. The data were expressed as mean ± standard error (Mean ± SEM). When P < 0.05, it was considered statistically significant. Figure 1b-Figure 1c As shown, the liver weight ratio of the mPGES-2 knockdown group was statistically significant compared with that of the control group (*P<0.05, **P<0.01).

[0050] Example 2: Effect of mPGES-2 knockout on liver regeneration after 70% liver resection in mice

[0051] Before the experiment, 12 8-week-old male ptges2 were taken. flox / flox Mice were randomly divided into two groups according to their body weight, namely a control group and an experimental group, and injected with a control virus and a Cre virus, respectively, to construct a control and mPGES-2 liver-specific knockout mouse model. The surgical method was the same as in Example 1, and the expression of mPGES-2 was detected by Western Blot using the same method as in Example 1. The expression results of mPGES-2 are shown in Figure 2a As shown. Figure 2b As shown, the liver weight ratio of the NC group was compared with that of the mPGES-2KO group, which was statistically significant (*P<0.05).

[0052] Example 3: Detection of biochemical indicators of liver function

[0053] (1) Plasma was collected from the NC group and mPGES-2KD mice 48 h and 96 h after liver resection, and plasma was collected from the NC group and mPGES-2KO mice 48 h after liver resection.

[0054] (2) Centrifuge at 3000 rpm for 15 min and collect serum.

[0055] (3) The levels of AST and ALT in serum were determined using AST and ALT detection kits (brand: Nanjing Jiancheng).

[0056] The results are as follows Figure 3a , Figure 3b , Figure 3c shown.

[0057] Example 4: Immunohistochemical staining

[0058] (1) Fixation of tissue specimens: The liver tissues of mice in each group were fixed in 4% paraformaldehyde at room temperature for 24 h, wrapped with gauze and marked accordingly, and then rinsed with running water overnight;

[0059] (2) Dehydration and transparency: Dehydrate in 50%, 60%, 70%, 80% and 90% alcohol gradient for 2 hours, then dehydrate in 95% alcohol and 100% alcohol-I / II / III for 1 hour each, and then soak in xylene-I / II for 30 minutes each;

[0060] (3) Wax immersion and embedding: Wax immersion was performed in a 58°C incubator. Wax immersion was performed with paraffin-I for 1.5 h and paraffin-II for 2 h. The samples were placed in an embedding box and embedded in paraffin at 60°C. After the samples were cooled and solidified into a block, the wax block was removed.

[0061] (4) Sectioning and spreading: Slice with a microtome to a thickness of 5 μm, then spread the slices in a 50°C water bath, mount the slices on a clean glass slide, and bake them in a 60°C oven overnight.

[0062] (5) Drying: Place the paraffin sections in a 60°C oven and dry for at least 1 hour.

[0063] (6) Dewaxing: Immerse the baked paraffin sections completely in xylene for dewaxing: soak in xylene I for 20 min and in xylene II for 20 min;

[0064] (7) Hydration: The dewaxed paraffin sections were completely immersed in different concentrations of ethanol for hydration: 100% ethanol for 10 min, 95% ethanol for 5 min, 80% ethanol for 5 min, 75% ethanol for 5 min, and tap water or PBS for several times;

[0065] (8) Antigen repair: Add an appropriate amount of sodium citrate antigen repair solution to a pressure cooker, immerse the rinsed paraffin sections in the sodium citrate antigen repair solution (the liquid level does not cover the tissue), place the pressure cooker in a microwave and heat for 10 minutes until the antigen repair solution boils, open the lid and check for bubbles (bubbles indicate that the sodium citrate antigen repair solution has boiled), cover the lid and continue heating for 5 minutes, then open the lid and cool naturally at room temperature, generally about 30 minutes;

[0066] (9) Rinse the repaired paraffin sections with PBS three times, 5 min each time;

[0067] (10) Blocking endogenous catalase: Use 3% hydrogen peroxide to completely immerse the paraffin sections, block them in the dark at room temperature for 30 min, and then rinse the paraffin sections with PBS three times, each time for 5 min;

[0068] (11) Blocking endogenous antigens: Use 5% BSA antigen blocking solution prepared with 0.1% PBST and block at room temperature for 60 min;

[0069] (12) Primary antibody incubation: add 0.1% PBS diluted primary antibody working solution, incubate at 4°C overnight, and rinse with PBS three times, 5 min each time;

[0070] (13) Secondary antibody incubation: add an appropriate amount of HRP-labeled secondary antibody working solution of the corresponding species and incubate at room temperature for 60 min;

[0071] (14) DAB color development: Prepare 1× DAB color development solution (brand: Zhongshan Jinqiao) according to the manufacturer's instructions for use of the reagent, add it dropwise to the dried paraffin tissue, react for a period of time, observe the color development under a microscope, terminate the staining with tap water in time, and rinse the paraffin sections with tap water several times;

[0072] (15) Hematoxylin counterstaining: Immerse the rinsed paraffin sections in hematoxylin stain for 10-20 seconds, then wash the hematoxylin stain with PBS, and then soak the paraffin sections in PBS with a pH of 7.2-7.4 for 10 minutes;

[0073] (16) Tissue dehydration: Tissue dehydration was performed as follows: 75% ethanol for 5 min, 80% ethanol for 5 min, 95% ethanol for 5 min, and 100% ethanol for 10 min;

[0074] (17) Paraffin wax clearing: Tissue clearing was performed as follows: xylene I for 20 min, xylene II for 20 min;

[0075] (18) Sealing: Add appropriate amount of resin to seal the slide, and be careful to remove all bubbles.

[0076] The results are as follows Figure 4 and Figure 5 shown.

[0077] Example 5: RNA-seq

[0078] (1) RNA extraction: 30 mg of regenerating liver tissue from the NC group and mPGES-2KO was used to extract total RNA. The concentration and purity of the extracted RNA were tested by Nanodrop2000, the integrity of RNA was tested by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. The total RNA amount required for single library construction was 10 ng, the concentration was ≥1 ng / μL, the RIN value was greater than 6.5, and the OD260 / 280 value was between 1.8 and 2.2.

[0079] (2) Reverse transcription into cDNA: Use Oligo(dT)primer to reverse transcribe RNA with poly A tail. Since a special active reverse transcriptase (Moloney Murine Leukemia Virus) is used for reverse transcription, three Cs will be added to the 3′ end of the cDNA chain.

[0080] (3) Template replacement: TSO (template-switching oligo) primers were used to synthesize the second strand of cDNA, thereby replacing the RNA complementary to the first strand of cDNA. It should be noted that the three Gs at the 3′ end of TSO can complement the three Cs at the 3′ end of the first strand, and the +G at the end is a modified G that can increase the thermal stability of TSO and its ability to complement the free 3′ end of the first strand of cDNA.

[0081] (4) PCR amplification: Mild cDNA enrichment, amplify cDNA to ng level.

[0082] (5) Fragmentation and adding adapters: The modified highly active Tn5 transposase is used to fragment the DNA and add adapters to both ends of the cDNA. The DNA fragments after labeling are usually 200-600 bp.

[0083] (6) Fragment screening and library enrichment: The products after adding adapters are purified and fragments are sorted, and the sorted products are used for PCR amplification and purification to obtain the final library.

[0084] (7) Sequencing on the NovaSeq X Plus platform: Qubit 4.0 quantification and sequencing; bridge PCR amplification to generate clusters; sequencing on the NovaSeq Xplus platform.

[0085] The experimental results are as follows Figure 6a , Figure 6b , Figure 6c , Figure 6d shown.

[0086] Experimental results:

[0087] (1) mPGES-2 liver-specific knockdown promotes liver regeneration after 70% liver resection: 70% liver resection was performed in the control and mPGES-2 liver-specific knockdown models, and liver regeneration was detected 48h and 96h after surgery. First, the expression of mPGES-2 in the liver tissue of mPGES-2 liver-specific knockdown mice was detected, showing that effective mPGES-2 liver-specific knockdown mice (such as Figure 1a ). Secondly, the regenerated liver weight and body weight of mice were measured and calculated using the formula "liver weight ratio = liver weight / body weight * 100%". The results showed that the liver weight ratio of the mPGES-2 liver-specific knockdown group was increased compared with the control group and was significantly different (*P < 0.05, **P < 0.01) (as shown in Figure 1b-Figure 1c This indicates that liver-specific knockdown of mPGES-2 increased the liver weight ratio at 48h and 96h after 70% hepatectomy and accelerated liver regeneration.

[0088] (2) mPGES-2 liver-specific knockout promotes liver regeneration after 70% liver resection: 70% liver resection was performed in the control and mPGES-2 liver-specific knockout models, and liver regeneration was detected 48 hours after surgery. First, the expression of mPGES-2 in the liver tissue of mPGES-2 liver-specific knockout mice was detected, showing that an effective mPGES-2 liver-specific knockout mouse (such as Figure 2a Secondly, the weight of the regenerated liver and the body weight of the mice were measured, and the liver weight ratio was calculated. The results showed that the liver weight ratio of the mPGES-2 liver-specific knockout group increased significantly compared with the control group (*P < 0.05), indicating that the liver weight ratio of mPGES-2 liver-specific knockout increased 48h after 70% liver resection and accelerated liver regeneration.

[0089] (3) Liver-specific knockdown or knockout of mPGES-2 does not affect regenerative liver function: To evaluate the effect of mPGES-2 inhibition on regenerative liver function, AST and ALT in mouse serum were detected. The results showed that neither mPGES-2 knockdown nor knockout had a significant effect on regenerative liver function indicators compared with the control group (e.g. Figure 3a , Figure 3b , Figure 3c ), indicating that liver-specific knockdown or knockout of mPGES-2 does not affect regenerative liver function.

[0090] (4) Liver-specific knockdown or knockout of mPGES-2 promotes hepatocyte proliferation and DNA replication in regenerating liver: Studies have shown that the success of liver regeneration is mainly attributed to the proliferation of parenchymal hepatocytes in the liver. This process is a key factor in the recovery of liver function. Ki-67 is a marker of cell proliferation and also a marker protein for liver regeneration. An increase in its content and the proportion of it entering the cell nucleus represents an accelerated rate of liver regeneration. Immunohistochemistry was used to detect the expression of Ki-67 in regenerating liver tissue, and it was found that liver-specific knockdown of mPGES-2 significantly increased the proportion of Ki-67 entering the cell nucleus (***P < 0.001) (such as Figure 4 ), indicating that liver-specific knockdown of mPGES-2 can promote hepatocyte proliferation in the regenerating liver;

[0091] DNA replication is a necessary prerequisite for the proliferation of regenerating hepatocytes. Therefore, immunohistochemistry was used to further detect the marker protein PCNA of DNA replication. The results showed that the proportion of PCNA positive cells in liver-specific knockout of mPGES-2 increased significantly (***P < 0.001) (e.g. Figure 5 This indicates that liver-specific knockout of mPGES-2 can promote the process of hepatocyte DNA replication in the regenerating liver.

[0092] (5) Liver-specific knockout of mPGES-2 promotes the expression of genes in the cell cycle and DNA replication pathways: To further elucidate the molecular mechanism by which liver-specific knockout of mPGES-2 affects hepatocyte proliferation and DNA replication, RNA-seq was performed on the regenerating liver tissues of mice in the NC group and the mPGES-2KO group. The results showed that multiple genes showed upregulation and downregulation after mPGES-2KO (e.g. Figure 6a ), indicating that there were differential gene expressions between the NC and KO groups after hepatectomy. KEGG enrichment analysis of the differentially expressed genes revealed that the cell cycle and DNA replication pathways were the most significantly different (e.g. Figure 6b The results showed that the transcription of differential genes involved in cell cycle and DNA replication was significantly upregulated in the mPGES-2KO group. The above results indicate that mPGES-2KO can promote the expression of genes in the cell cycle and DNA replication pathways, thereby promoting DNA replication and the proliferation of regenerative hepatocytes.

[0093] In summary, through the above experimental comparison, the role of mPGES-2 in the liver regeneration process was deeply explored. Through liver-specific gene knockdown or knockout experiments on mPGES-2, it was found that inhibiting mPGES-2 can promote liver regeneration. Specifically, after liver-specific knockout of mPGES-2, the expression of genes involved in the cell cycle and DNA replication was significantly upregulated, thereby promoting the proliferation of hepatocytes, ultimately leading to an increase or maintenance of liver volume and function. This series of results shows that mPGES-2 has the potential to become an important target for liver regeneration, providing useful reference and guidance for clinical drug development in the field of liver regeneration.

[0094] In addition, the inventors of this case also referred to the aforementioned embodiments and conducted experiments with other raw materials, process operations, and process conditions described in this specification, and obtained relatively ideal results.

[0095] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements all fall within the scope of the present invention to be protected.

Claims

1. Application of mPGES-2 as a target in the development, screening or preparation of drugs for promoting liver regeneration.

2. The use according to claim 1, characterized in that: The liver regeneration refers to the liver regeneration phenomenon after 70% of the liver is removed.

3. The use according to claim 1, characterized in that: When the drug acts on a mouse model, it can at least inhibit the expression of mPGES-2 in the mouse model, and the mouse model is a mouse subjected to 70% partial liver resection.

4. The use according to claim 3, characterized in that: When the drug acts on a mouse model, it can at least increase the regenerated liver weight ratio.

5. The use according to claim 3, characterized in that: When the drug acts on a mouse model, it can at least promote the expression of proliferation antigen Ki-67 in regenerating liver cells; And / or, when the drug acts on a mouse model, it can at least promote the expression of PCNA, a DNA replication marker, in regenerating liver cells; and / or, when the drug acts on a mouse model, it can at least promote the expression of cell cycle genes in regenerating hepatocytes; And / or, when the drug acts on a mouse model, it can at least promote the expression of DNA replication genes in regenerating hepatocytes.

6. The use according to claim 3, characterized in that: When the drug acts on a mouse model, it can at least promote the expression of cell proliferation protein and DNA replication protein in regenerating liver cells.

7. Use of mPGES-2 or an inhibitor of its encoding gene in the preparation of a drug for promoting liver regeneration.

8. The use according to claim 7, characterized in that: The inhibitor is selected from interfering molecules that specifically interfere with the expression of the gene encoding mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its encoding gene.

9. A pharmaceutical composition for promoting liver regeneration, characterized in that: include: An inhibitor of mPGES-2 or its encoding gene, and a pharmaceutically acceptable carrier.

10. The pharmaceutical composition according to claim 9, characterized in that: The inhibitor is selected from interfering molecules that specifically interfere with the expression of the gene encoding mPGES-2 and / or small molecule compounds that specifically inhibit mPGES-2 or its encoding gene.

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