New uses of fam177a1

Abstract

The application belongs to the field of FAM177A1 application research, and particularly relates to a new application of FAM177A1. Mainly disclosed are: application of a preparation for detecting FAM177A1 in preparation of a detection product for a myocardial cell hypertrophy disease; application of FAM177A1 or an expression promoter thereof in preparation of a medicine for treating the myocardial cell hypertrophy disease; application of FAM177A1 in preparation of a medicine for treating a disease induced by activation of a MAPK / ERK channel; and application of FAM177A1 in preparation of an inhibitor for non-diagnostic treatment activity of a MAPK enzyme. In the application, FAM177A1 has an inhibiting effect on MAPK, FAM177A1 can become a target for resisting cardiac hypertrophy, so as to prevent or treat occurrence and development of cardiac hypertrophy; and FAM177A1 can also be used as a product for diagnosing and treating cardiac hypertrophy caused by peripheral vascular resistance such as aortic arch constriction and hypertension.

Description

Technical Field

[0001] This invention belongs to the field of FAM177A1 application research, and in particular relates to new uses of FAM177A1. Background Technology

[0002] Sequence similarity family 177 member A1, English name FAM177A1, also known as C14orf24; Chromosome14 Open Reading Frame 24; contains 236 amino acids, with a molecular weight of 23960 Da. NCBI reference sequence number: NP_775878.2, amino acid sequence as follows:

[0003] MEVGLPAITLFLTSASSPVVATTMDQEPVGGVERGEAVAASGAAAAAAFGESAGQMSNERGFENVELGVIGKKKKVPRRVIHFVSGETMEEYSTDEDEVDGLEKKDVLPTVDPTKLTW GPYLWFYMLRAATSTLSVCDFLGEKIASVLGISTPKYQYAIDEYYRMKKEEEEEEENRMSEEAEKQYQQNKLQTDSIVQTDQPETVISSSFVNVNFEMEGDSEVIMESKQNPVSVPP.

[0004] In recent years, research on Family With Sequence Similarity (FAM) family members regarding tumor progression and metabolic cardiovascular diseases has attracted significant attention. These metabolic cardiovascular diseases include metabolic syndrome, diabetes, and myocardial hypertrophy. FAM177A1 is a newly discovered member of the FAM family with an unknown function, inhibiting IL-1β-activated NF-κB and inflammatory gene transcription. Its role in cardiovascular and metabolic-related diseases remains unclear. Our previous studies found that FAM177A1 significantly inhibited cardiomyocyte hypertrophy caused by aortic arch constriction, delaying the development of cardiac hypertrophy. We also found that FAM177A1 reduced the incidence of hypertension caused by phenylephrine (PE)-induced peripheral vascular resistance. Therefore, we hypothesize that FAM177A1 may play an important role in cardiac biology, particularly in the regulation of cardiac dysfunction caused by peripheral vascular resistance. Currently, it remains unclear whether FAM177A1 has the potential to be used as a drug for the prevention and treatment of cardiac hypertrophy.

[0005] There are many causes of cardiac hypertrophy, including hypertensive left ventricular hypertrophy, congestive heart failure, anterior wall myocardial infarction, mitral regurgitation, aortic stenosis, hypertrophic cardiomyopathy, pulmonary hypertension, cor pulmonale, dilated cardiomyopathy, endocarditis, left ventricular aneurysm, and mitral stenosis. Cardiac hypertrophy can be divided into two main categories: enlargement and hypertrophy. ① Enlargement refers to a condition where the heart muscle does not thicken; sometimes, the myocardium even thins. This condition is usually caused by the heart muscle losing its original elasticity, leading to increased volume and pressure, resulting in heart enlargement. Common causes include myocarditis, valvular insufficiency, hyperthyroidism, and vitamin B1 deficiency. ② Hypertrophy refers to a condition where the heart muscle thickens, but the heart chambers do not enlarge; sometimes, they even shrink, but the overall appearance of the heart is larger than normal. Common causes of cardiac hypertrophy include narrowing of the heart outlet, such as aortic stenosis; increased peripheral vascular resistance, such as hypertension; and cardiomyopathy, such as hereditary hypertrophic cardiomyopathy. The main danger of cardiac hypertrophy is that the heart's contractility decreases after it enlarges, thus leading to heart failure.

[0006] Currently, anti-cardiac hypertrophy methods are mainly divided into three categories: ① Reversing cardiac hypertrophy: Commonly used drugs include angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, beta-blockers, and aldosterone receptor antagonists, which reverse cardiac chamber enlargement to some extent through neuro-humoral regulation; ② Reducing cardiac load: Correcting heart failure by reducing cardiac load. Treatment methods include diuresis and vasodilators, with loop diuretics being the most commonly used; ③ Using vasodilators: ACEIs (angiotensin-converting enzyme inhibitors), ARBs (angiotensin II receptor blockers), and nitrates. The role of FAM177A1, a newly discovered protein, in cardiac hypertrophy remains unstudied.

[0007] Mitogen-activated protein kinase (MAPK) is a group of serine-threonine protein kinases that can be activated by various extracellular stimuli, such as cytokines, neurotransmitters, hormones, cellular stress, and cell adhesion. Several inhibitors targeting MAPK activity have been developed, such as PD98059, the flavonoid Scutellarin, and Selumetinib (AZD6244). The regulatory role of FAM177A1, a newly discovered protein, in mitogen-activated protein kinase has not yet been studied. Protease inhibitors are new products developed in the mid-to-late 1990s. Their complex synthesis processes are a major reason for the high cost of anti-HIV treatment, and they are often referred to as "barriers that must be overcome." Common protease inhibitors include nelfinavir, saquembavir, indinavir, amprenavir, ritonavir, lopinavir, and combination formulations. Summary of the Invention

[0008] To address the aforementioned issues, this invention provides new applications for FAM177A1, primarily filling the gaps in the application of FAM177A1 in the diagnosis and treatment of certain diseases, and also filling the gaps in the application of FAM177A1 in protein regulation.

[0009] To solve the above problems, the present invention adopts the following technical solution:

[0010] Application of formulations for detecting FAM177A1 in the preparation of diagnostic products for cardiomyocyte hypertrophy.

[0011] In some embodiments, the preparation for detecting FAM177A1 is a FAM177A1 expression assay; the cardiomyocyte hypertrophy disease includes myocardial hypertrophy and cardiac hypertrophy.

[0012] In some embodiments, cardiac hypertrophy includes ventricular hypertrophy, cardiac enlargement, and atrial enlargement.

[0013] The cardiac hypertrophy mentioned above refers to hypertrophic cardiomyopathy induced by increased peripheral vascular resistance (such as hypertension), narrowing of the heart outlet (such as aortic stenosis), or cardiomyopathy (such as hereditary hypertrophic cardiomyopathy).

[0014] Application of FAM177A1 or its expression promoter in the preparation of drugs for treating cardiomyocyte hypertrophy.

[0015] Application of FAM177A1 in the preparation of drugs for treating diseases induced by MAPK / ERK pathway activation.

[0016] In some embodiments, FAM177A1 is overexpressed, and the expression promoter is a FAM177A1 overexpression promoter; the cardiomyocyte hypertrophy disease is myocardial hypertrophy or cardiac hypertrophy.

[0017] In some embodiments, cardiac hypertrophy includes ventricular hypertrophy, cardiac enlargement, and atrial enlargement.

[0018] The cardiac hypertrophy mentioned above refers to hypertrophic cardiomyopathy induced by increased peripheral vascular resistance (such as hypertension), narrowing of the heart outlet (such as aortic stenosis), or cardiomyopathy (such as hereditary hypertrophic cardiomyopathy).

[0019] In some embodiments, the FAM177A1 or its expression promoters are characterized by inhibiting cardiomyocyte hypertrophy and collagen fiber increase.

[0020] Application of FAM177A1 in the preparation of drugs for the prevention and treatment of hypertension (including hypertension caused by peripheral vascular resistance due to PE).

[0021] Application of FAM177A1 in the preparation of non-diagnostic therapeutic activity inhibitors of MAPK enzymes or MAPK / ERK kinases or / ERK kinases.

[0022] In some embodiments, FAM177A1 is overexpressed.

[0023] The beneficial effects of this invention are:

[0024] FAM177A1 inhibits MAPK and can serve as a target for anti-cardiac hypertrophy, thereby preventing or treating the occurrence and development of cardiac hypertrophy. FAM177A1 can also be used as a product for diagnosing and treating peripheral vascular resistance such as aortic arch coarctation and cardiac hypertrophy caused by hypertension. Attached Figure Description

[0025] Figure 1 PE treatment was used to express FAM177A1 protein in primary rat cardiomyocytes.

[0026] Figure 2 Gene expression of myocardial hypertrophy-related factors (ANP, BNP) in the control and treatment groups;

[0027] Figure 3 Western blot and qPCR were used to detect protein / gene expression in primary rat myocardial cells in the sham and TAC groups.

[0028] Figure 4 MASSON staining was used to detect cardiac hypertrophy in both the control and treatment groups.

[0029] Figure 5 MASSON staining was used to detect the expression of cardiac collagen fibers (blue) and muscle fibers (red) in the control and treatment groups.

[0030] Figure 6 This refers to the expression of molecular proteins and enzymes in the MAPK signaling pathway. Detailed Implementation

[0031] The technology of the present invention will be further described below:

[0032] The first aspect of this section introduces the application of FAM177A1 in the diagnosis of cardiomyocyte hypertrophy. :

[0033] Application of formulations for detecting FAM177A1 in the preparation of diagnostic products for cardiomyocyte hypertrophy.

[0034] FAM177A1 is positively correlated with the occurrence of hypertrophic cardiomyopathy. This invention aims to detect hypertrophic cardiomyopathy by detecting FAM177A1 and to correlate the development of hypertrophic cardiomyopathy with FAM177A1. The aforementioned formulations for detecting FAM177A1 are not limited to reagents (such as probes, primers, etc.); other methods that can achieve the purpose of detecting FAM177A1 and are used to evaluate hypertrophic cardiomyopathy should also be included within the scope of this invention, including new and feasible technologies.

[0035] One of these is the cardiomyocyte hypertrophy disease, which includes myocardial hypertrophy and cardiac hypertrophy. The cardiac hypertrophy includes ventricular hypertrophy, cardiac enlargement, and atrial enlargement.

[0036] Hypertrophic cardiomyopathy has multiple precipitating types, including hypertrophic cardiomyopathy induced by hypertension, aortic stenosis, or increased intraventricular pressure; the detection products include diagnostic products.

[0037] Secondly, the preparation used to detect FAM177A1 is a FAM177A1 expression detection reagent. Detection reagents include PCR and Western blotting (WB) assays. The expression level of FAM177A1 in the sample is detected using PCR technology. When the sample is a sample of a corresponding disease, its FAM177A1 expression will also increase accordingly. The specific standard for the increase in expression level is based on conventional medical standards.

[0038] The second aspect of this section introduces the application of FAM177A1 in the prevention and treatment of cardiomyocyte hypertrophy. :

[0039] First, the application of FAM177A1 or its expression promoters in the preparation of drugs for treating hypertrophic cardiomyocyte disease. FAM177A1 or its expression promoters serve as the main active ingredient in the preparation of drugs for treating hypertrophic cardiomyocyte disease.

[0040] The aforementioned cardiomyocyte hypertrophy disease includes myocardial hypertrophy and cardiac hypertrophy. The cardiac hypertrophy includes ventricular hypertrophy, cardiac enlargement, and atrial enlargement.

[0041] Meanwhile, the aforementioned cardiomyocyte hypertrophy diseases include hypertrophic cardiomyopathy induced by hypertension or increased intraventricular pressure. Cardiomyocyte hypertrophy diseases also include complications caused by cardiomyocyte hypertrophy, such as endocarditis, heart failure, arterial embolism, and arrhythmias induced by cardiomyocyte hypertrophy. By slowing down cardiomyocyte hypertrophy, the complications can also be treated accordingly.

[0042] Second, the application of FAM177A1 in the preparation of drugs for treating diseases induced by MAPK / ERK pathway activation. FAM177A1 treats the corresponding diseases by inhibiting MAPK / ERK pathway activation. Diseases induced by MAPK / ERK pathway activation include cardiomyocyte hypertrophy, as well as other induced diseases that can be treated and alleviated by inhibiting MAPK / ERK.

[0043] Among these, the induced diseases that can be treated and alleviated by inhibiting MAPK / ERK are included. Besides cardiomyocyte hypertrophy, the aforementioned induced diseases can also be interpreted as diseases that have been proven to be treatable by inhibiting MAPK / ERK prior to the application date of this invention, including at least the following related diseases. For example, inhibiting MAPK / ERK to combat skin aging and radiation-induced premature skin aging (CN113853218A), secretory curl-related protein 1 inhibiting the migration and invasion of colorectal cancer cells through the MAPK / ERK signaling pathway (Anhui Medical Journal), and exploring the mechanism of warming the meridians, removing blood stasis, and relieving pain based on the MAPK / ERK signaling pathway in treating primary dysmenorrhea due to cold stagnation and blood stasis (Chinese Journal of Experimental Traditional Medical Formulae), etc., should also fall under the category of diseases treatable by FAM177A1 pharmaceutical as described in this article. Specific verification experiments will not be elaborated upon in this invention.

[0044] In pharmaceutical applications, FAM177A1 is primarily used as a main active ingredient in drugs, and it is used as a component in the pharmaceutical manufacturing process.

[0045] In some instances, FAM177A1 is overexpressed, such as in a FAM177A overexpressing adenovirus. The FAM177A1 expression promoter is any FAM177A1 overexpression promoter; no specific limitations are imposed, and any existing promoter can be used.

[0046] In drug therapy, FAM177A1 or its expression promoters inhibit cardiomyocyte hypertrophy and collagen fiber increase, thereby achieving the treatment of the corresponding condition.

[0047] The third aspect of this section introduces the application of FAM177A1 in non-diagnostic and therapeutic scenarios. :

[0048] Applications of FAM177A1 in Non-Diagnostic Therapeutic Inhibitors of MAPK Enzymes. This application primarily targets non-disease diagnosis and treatment scenarios, and does not fall under the pharmaceutical field. It mainly manifests in some biological modulatory agents, such as some commercial inhibitors. For example, FAM177A1 can be used as an in vitro activity inhibitor of any of the following enzymes: MAPK enzyme, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), MAPK / ERK kinase, ERK enzyme, Jnk enzyme, and p38 enzyme, thereby achieving partial control of the experimental environment. In non-diagnostic treatment, FAM177A1 can be used either as an active ingredient in inhibitors or directly as a standalone inhibitor. This includes using FAM177A1 to prepare inhibitors, as well as using FAM177A1 directly as a non-diagnostic therapeutic inhibitor of MAPK enzymes; both fall under the applications described in this section.

[0049] Furthermore, in the front-end use environment, inhibiting MAPK activity includes inhibiting multiple of its signaling pathways.

[0050] The FAM177A1 mentioned above can be an overexpression of FAM177A1, and the use of overexpression of FAM177A1 can significantly inhibit the expression of related substances.

[0051] The fourth aspect of this section introduces specific research projects. :

[0052] Experimental content

[0053] Masson staining assay: Masson staining involves mixing two or three anionic dyes, resulting in blue staining for collagen fibers and red staining for muscle fibers. It is one of the staining methods used to visualize fibers and inflammatory factors in tissues. Heart-specific supergene FAM177A1 (FAM) rats and control rats (GFP) underwent sham surgery (SHAM) and aortic arch coarctation surgery (TAC), respectively. Eight weeks post-surgery, rat heart tissue was isolated and analyzed using Masson staining (Bio-Aus Biotech, Wuhan, Hubei). Results were captured using OlympuscellSens Entry.

[0054] PE-induced cardiomyocyte hypertrophy assay: Primary rat cardiomyocytes were seeded in 6-well plates and cultured to 80% confluence. Cells were treated with solvent control and PE (50 μM) for 24 h, respectively. Cells were then collected, and changes in cardiomyocyte hypertrophy markers were detected using Western blotting (WB) or quantitative real-time PCR (qPCR).

[0055] Transverse aortic constriction (TAC) experiment in rats: TAC is used to simulate hypertrophic cardiomyopathy caused by hypertension or increased intraventricular pressure. After exposing the rat aortic arch, a suture is threaded through the arch and knotted, with a constriction needle placed inside the knot. After tightening the suture, the needle is removed, the thoracic cavity is closed, and the skin of the neck and chest is sutured. Changes in corresponding indicators of myocardial hypertrophy are detected at 1, 2, 4, and 8 weeks postoperatively, and the heart is photographed using the Olympus CellSens channel.

[0056] Adenovirus packaging and transfection: 293 cells were seeded in a 60mm culture dish. When the cell density reached 50-70%, 6μg of plasmid was transfected into the cells via PEI. After culturing for 6-8 hours, the culture medium was removed, and 2-3ml of DMEM complete medium (10% FBS by volume) was added. 7-10 days post-transfection, the cells were pipetted off. The solution was collected, centrifuged, and the supernatant was discarded. The cells were resuspended in 2.0ml PBS, frozen in liquid nitrogen, and thawed in a 37°C water bath with vigorous shaking. This step was repeated three times to obtain a virus-containing supernatant. 293 cells were seeded at a confluence of 50-70% in a 60mm culture dish, and the virus-containing supernatant was added at a volume ratio of 30-50%. Obvious cell lysis or cytopathic effect (CPE) was observed 2-3 days post-infection, and the virus supernatant was collected again. Three rounds of amplification were performed to obtain a sufficient viral titer. Rat primary cardiomyocytes were cultured to a cell density of 70-80%, and the collected virus was used to infect the rat primary cardiomyocytes. After 24-48 hours, the cell fluorescence was observed under a microscope.

[0057] In vitro activity regulation experiments of MAPK enzyme, ERK kinase, etc.: The in vitro protein incubation experiment used rat-derived FAM177A1 protein at a concentration of 0.1 μg / ml to verify the expression of MAPK enzymes under the action of FAM177A1.

[0058] Results Analysis

[0059] Result 1 After PE-induced myocardial hypertrophy, the expression of ANP and FAM177A1 increased, and the expression of FAM177A1 was positively correlated with PE-induced myocardial hypertrophy.

[0060] Figure 1 In the middle a, the myocardial progenitor cells of male SD rats were cultured and treated with control (Vehicle) and PE (50 μM) for 24 h, respectively. Figure 1 In Figure b, rat primary cardiomyocytes were treated with different concentrations of PE (5, 50, and 150 μM). Histones from different treatments were collected, and the expression levels of FAM177A1 and ANP (a marker of myocardial hypertrophy) proteins were detected by Western blotting.

[0061] Depend on Figure 1 As shown in a, compared with the control group, PE treatment induced an increase in FAM177A1 expression in primary rat myocardial cells; Figure 1 As shown in Figure b, the expression of FAM177A1 in primary rat myocardial cells increased with the increase of PE treatment concentration, and the expression of the myocardial hypertrophy marker ANP also increased, suggesting that the expression of FAM177A1 increased with the increase of PE-induced myocardial cell hypertrophy, and that the expression of FAM177A1 was positively correlated with myocardial hypertrophy.

[0062] It was also verified that FAM177A1 can reduce the incidence of hypertension caused by peripheral vascular resistance induced by phenylephrine (PE) (the specific verification method is similar to that in the previous section, and will not be repeated here). The aforementioned hypertension is PE-induced hypertension.

[0063] Result 2 Intervention in FAM expression regulates cardiomyocyte hypertrophy.

[0064] Figure 2 Rat primary myocardial cells were treated with control (Adv-GFP), FAM177A1-over-adenovirus (Adv-FAM), and FAM177A1-knockdown adenovirus (Adv-sh-FAM) for 24 h with phosphate-buffered saline (PBS) and PE (50 μM). Cells were then collected, RNA was extracted, and qPCR was used to detect myocardial hypertrophy-related markers (ANPs). Figure 2 a) BNP ( Figure 2 b)) and the cardiomyocyte-specific expression index MYH7 ( Figure 2 c)) Expression level.

[0065] Depend on Figure 2 It can be seen that, under no-stimulation conditions, overexpression or knockdown of FAM slightly increased the expression of ANP and BNP in cardiomyocytes. Under PE-induced stimulation, overexpression of FAM significantly inhibited the expression of ANP and BNP, while knockdown of FAM led to a significant increase in the expression of myocardial hypertrophy markers. Figure 2 a) and b); while PE and / or intervention in FAM expression had no similar effect on the expression of the cardiomyocyte marker MYH7. These results indicate that in the PE-induced cardiomyocyte hypertrophy model, intervention in FAM expression regulates cardiomyocyte hypertrophy.

[0066] Result 3 TAC surgery led to increased expression of ANP, BNP, and FAM177A1 in rats with myocardial hypertrophy, and FAM177A1 expression was positively correlated with the degree of myocardial hypertrophy.

[0067] After rats underwent sham surgery or TAC surgery, the expression of cardiomyocyte-related indicators was detected by Western blotting and qPCR.

[0068] Figure 3 In the figure, 'a' represents the expression of FAM177A1 and β-tubulin proteins in the heart at 8 weeks post-TAC (8 weeks after surgery) using Western blotting. The first column shows FAM177A1 expression, and the second column shows the expression of the internal reference protein β-tubulin.

[0069] The levels of fam177a1 in the hearts of rats in the sham-operated group, 1 week after TAC (TAC 1w), 2 weeks after TAC (TAC 2w), 4 weeks after TAC (TAC 4w), and 8 weeks after TAC (TAC 8w) were detected by qPCR. Figure 3 b) ANP ( Figure 3 c) BNP ( Figure 3 d), MYH6 Figure 3 e).

[0070] Depend on Figure 3 As shown in a and b, TAC surgery in rats significantly increased both the protein and RNA levels of FAM177A1; Figure 3 As shown in C and D, TAC surgery leads to an increase in the expression of myocardial hypertrophy markers ANP and BNP, while the expression of the cardiomyocyte-specific marker MYH6 decreases with prolonged time after TAC surgery. Figure 3 e). The results showed that TAC surgery induced myocardial hypertrophy in rats, and FAM177A1 expression was positively correlated with the degree of myocardial hypertrophy.

[0071] Result 4 FAM177A1 significantly inhibited TAC surgery-induced cardiac hypertrophy and ventricular enlargement in rats.

[0072] Figure 4 FAM177A1 heart-specific hyperplasia rats (FAM) and control rats (GFP) underwent sham surgery (SHAM) and TAC surgery, respectively. Masson staining was performed on the transverse sections of the heart 8 weeks later.

[0073] Figure 4 In the middle section (a), the heart cross-section of a control rat undergoing sham surgery (GFP+SHAM group) is shown. Figure 4 In the middle section (b), the heart cross-section of a control rat undergoing TAC (GFP+TAC group) is shown. Figure 4 In the middle (c), the heart cross-section of FAM-treated rats after sham surgery (FAM+SHAM group) is shown. Figure 4 In the middle section, d represents the heart cross-section of FAM-exposed rats undergoing TAC (FAM+TAC group).

[0074] Depend on Figure 4 As shown in a and b, TAC surgery leads to cardiac hypertrophy and ventricular enlargement in rats.

[0075] Depend on Figure 4 As shown in b and d, FAM177A1 significantly inhibited TAC surgery-induced cardiac hypertrophy and ventricular enlargement in rats.

[0076] Result 5 FAM177A1 significantly inhibited TAC surgery-induced cardiomyocyte hypertrophy and collagen fiber increase in rats.

[0077] Figure 5 FAM177A1 heart-specific hyperexpression rats (FAM) and control rats (GFP) underwent sham surgery (SHAM) and TAC surgery, respectively. Eight weeks later, Masson staining was used to detect the expression of cardiac collagen fibers (blue, with arrows indicating parts) and myofibrils (red).

[0078] Figure 5 In figure a, the expression of cardiac collagen fibers and muscle fibers in control rats after sham surgery (GFP+SHAM group) is shown. Figure 5 In figure b, the expression of cardiac collagen fibers and muscle fibers in control rats subjected to TAC (GFP+TAC group) is shown. Figure 5 In the middle (c), the expression of cardiac collagen fibers and muscle fibers in FAM-overexpressing rats after sham surgery (FAM+SHAM group) is shown. Figure 5 In the middle d, the expression of cardiac collagen fibers and muscle fibers in FAM-overexpressing rats undergoing TAC (FAM+TAC group) is shown.

[0079] Depend on Figure 5 As shown in a and b, TAC surgery leads to hypertrophy of rat cardiomyocytes and an increase in collagen fibers.

[0080] Depend on Figure 5 As shown in b and d, FAM177A1 significantly inhibited TAC surgery-induced cardiomyocyte hypertrophy and collagen fiber increase in rats.

[0081] Result 6 FAM177A1 can directly inhibit the PE-induced MAPK / ERK signaling pathway and further inhibit cardiomyocyte hypertrophy and cardiac hypertrophy through its mechanism. It can also inhibit the activity of enzymes such as ERK in vitro.

[0082] Rat primary myocardial cells were treated with adenovirus control (GFP), FAM177A1 overexpressing adenovirus (Adv), and FAM177A1 knockdown adenovirus for 24 h, respectively, and then treated with PBS or PE (50 μM) for 24 h. Changes in the MAPK signaling pathway were detected by Western blotting.

[0083] Figure 6 The first column in column a represents phosphorylated jnk (P-jnk) expression, the second column represents total jnk (t-jnk) expression, the third column represents phosphorylated cjun (P-cjun) expression, and the fourth column represents the expression of the internal control protein GAPDH.

[0084] Figure 6 In column b, the first column shows phosphorylated erk (P-erk) expression, the second column shows total erk (t-erk) expression, the third column shows phosphorylated p38 (P-p38) expression, the fourth column shows total p38 (t-p38) expression, and the fifth column shows the expression of the internal control protein GAPDH.

[0085] Depend on Figure 6 As shown in a and b, PE promotes increased phosphorylation of MAKP / ERK, while overexpression of FAM177A1 significantly inhibits PE-induced activation of the MAPK / ERK signaling pathway in primary rat cardiomyocytes; conversely, inhibiting FAM177A1 expression significantly promotes activation of the MAPK / ERK signaling pathway.

[0086] Depend on Figure 6 In the in vitro protein incubation experiment, the protein used was rat-derived FAM177A1 at a concentration of 0.1 μg / ml. This indicates that FAM177A1 can also directly play a regulatory role at the protein level, verifying the inhibitory effect of non-disease environments on the activities of MAPK enzymes, MAPK / ERK kinases, ERK enzymes, Jnk enzymes, and p38 enzymes. It can be directly used as an enzyme activity inhibitor for industrial applications, especially as a commercial experimental preparation.

[0087] In summary, the experimental results show that FAM177A1 can directly inhibit the PE-induced MAPK / ERK signaling pathway, thereby suppressing cardiomyocyte hypertrophy and cardiac hypertrophy. Furthermore, FAM177A1 can directly regulate activity, validating the feasibility of in vitro enzyme activity and peptide expression inhibitors.

[0088] Those skilled in the art will appreciate that various modifications to the above embodiments can be made without departing from the overall spirit and concept of the present invention. All such modifications fall within the protection scope of the present invention. The protection scheme of the present invention is defined by the appended claims.

Claims

1. Application of formulations for detecting FAM177A1 in the preparation of detection products for myocardial hypertrophy and cardiac hypertrophy induced by increased peripheral vascular resistance, narrowing of the cardiac outlet, or cardiomyopathy.

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