Method, system and equipment for evaluating treatment effect of anti-aging drug Senlytics

Abstract

The invention discloses a method, a system and equipment for evaluating the treatment effect of an anti-aging drug Senlytics. According to the invention, it is found for the first time that the anti-aging drug Senlytics composed of dasatinib and quercetin plays an anti-aging role by inhibiting the level of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 compounds, and on the basis of this, the anti-aging effect of the anti-aging drug Senlytics composed of dasatinib and quercetin is achieved. The invention provides a method, a system and equipment for evaluating the treatment effect of an anti-aging drug Senlytics composed of dasatinib and quercetin based on the expression level of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 compounds for the field, and the method, the system and the equipment have wide application prospects in the aspect of evaluation of the treatment effect of cardiovascular disease drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of bioinformatics, and specifically, the present application relates to a method, system and device for evaluating the therapeutic effect of Senolytics, an anti-aging drug. BACKGROUND

[0002] Cardiovascular disease (CVD) is one of the leading causes of morbidity and mortality worldwide. With the global population aging and lifestyle changes, the incidence of cardiovascular disease continues to rise, causing a huge burden on public health and social economic systems. Increasing evidence suggests that vascular aging is a key risk factor for the onset of cardiovascular disease. Vascular aging refers to the degenerative changes in the structure and function of blood vessels with age, characterized by increased arterial stiffness and increased pulse wave velocity. The underlying mechanisms mainly involve the aging of vascular endothelial cells and smooth muscle cells, as well as the accumulation of pro-inflammatory factors caused by persistent sterile inflammation. These changes lead to vascular remodeling, excessive deposition of collagen, and ultimately vascular fibrosis and atherosclerosis. Among these factors, the aging of vascular endothelial cells (ECs) plays a key role in the process of vascular aging and significantly accelerates the occurrence and development of cardiovascular disease. Vascular endothelial cells are a single layer of flat squamous cells lining the vascular lumen, serving as a barrier between blood and vascular wall / tissue. They regulate vascular tone and balance by producing and releasing vasoactive substances such as vasodilator nitric oxide (NO) and vasoconstrictor angiotensin II (Ang II). Endothelial cell aging impairs vascular function, promotes thrombosis, enhances oxidative stress, and accelerates the development of atherosclerosis. These changes weaken the self-regulatory ability of blood vessels, thereby exacerbating vascular aging.

[0003] Currently, drug treatments targeting vascular aging mainly include statins, cyclooxygenase 2 (COX-2) inhibitors (such as aspirin), calcium channel blockers, and sodium-glucose co-transporter 2 (SGLT2) inhibitors. These drugs delay the decline in vascular function by increasing the bioavailability of NO, inhibiting oxidative stress, improving endothelium-dependent vasodilation, and modulating inflammatory pathways. However, the efficacy of these drugs is often limited, and there are adverse side effects, so there is an urgent need for safer and more effective treatment strategies. In recent years, emerging anti-aging therapies targeting selective elimination of senescent cells have shown good therapeutic potential. Among various senolytic therapies, the combination of dasatinib and quercetin (D+Q) is considered the most representative. Studies have shown that D+Q not only effectively eliminates senescent cells, but also reduces the burden of naturally occurring senescent cells and their pro-inflammatory secretions associated with frailty in human adipose tissue explants, ultimately extending the healthy lifespan of animal models. However, the exact mechanism of D+Q in regulating endothelial cell aging remains to be elucidated. SUMMARY

[0004] In order to make up for the deficiencies of the prior art, the purpose of the present application is to elucidate the mechanism of D+Q regulating endothelial cell senescence, and further to screen biomarkers that can be used to evaluate the anti-aging effect of D+Q.

[0005] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0006] The first aspect of the present application provides a method for evaluating the treatment effect of anti-aging drug Senolytics.

[0007] Further, the anti-aging drug Senolytics is a drug combination of dasatinib and quercetin.

[0008] Further, the method is completed by a computer, and the method comprises the following steps:

[0009] Obtaining data: obtaining expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex in a blood vessel aging patient sample after treatment with anti-aging drug Senolytics;

[0010] Processing data: inputting the expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex into an evaluation model constructed based on the expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex to predict the treatment effect of anti-aging drug Senolytics;

[0011] Outputting results.

[0012] In the present application, "anti-aging drug Senolytics", "a drug combination of dasatinib and quercetin" and "D+Q" are synonymous, all referring to an anti-aging drug composed of dasatinib and quercetin.

[0013] In the present application, "p-p65 / p65" refers to the proportion of phosphorylated p65 to total p65.

[0014] Further, the construction steps of the evaluation model are as follows:

[0015] obtaining expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex from untreated vascular aging patients and vascular aging patients after treatment with anti-aging drug Senolytics; inputting the expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex into a machine learning algorithm to build an evaluation model.

[0016] Further, the machine learning algorithm includes an algorithm model developed by various development tools.

[0017] Further, the development tools include but are not limited to TensorFlow, Scikit Learn, PyTorch, OpenNN, RapidMiner, Azure Machine Learning, Apache Mahout, Shogun, KNIME, Vertex AI, H2Oai, Anaconda, Keras, Tableau, Fast.ai, Catalyst, Amazon ML, MLJAR, and Spell.

[0018] Further, the algorithm model includes but is not limited to linear regression model, logistic regression model, Lasso regression model, Ridge regression model, linear discriminant analysis model, nearest neighbor model, decision tree model, perceptron model, neural network model, support vector machine model, naive Bayes model, AdaBoost model, GBDT model, XGBoost model, LightGBM model, CatBoost model, and random forest model.

[0019] Further, the evaluation model obtains results by the following standards:

[0020] When the expression amount of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex is lower than the threshold value, the classification result that anti-aging drug Senolytics is effective for treating vascular aging patients is obtained; when the expression amount of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex is higher than the threshold value, the classification result that anti-aging drug Senolytics is ineffective for treating vascular aging patients is obtained.

[0021] In some embodiments, the preset threshold value is a representative value of a normal sample of a blood vessel aging population, including but not limited to a maximum value, a third quartile, an average value. In some preferred embodiments of the present application, the population sample includes more than 20 samples, for example, 30, 50, 80, 100, 150, 200, 300, 500 or more.

[0022] The second aspect of the present application provides a system for evaluating the treatment effect of the anti-aging drug Senolytics.

[0023] Further, the anti-aging drug Senolytics is a drug combination of dasatinib and quercetin.

[0024] Further, the system comprises:

[0025] A data acquisition unit for acquiring expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex in a blood vessel aging patient sample after treatment with the anti-aging drug Senolytics;

[0026] A data classification unit for classifying and predicting the data obtained in the data acquisition unit by the evaluation model obtained by the construction method of the first aspect of the present application, to obtain a classification result of whether the anti-aging drug Senolytics is effective for the treatment of blood vessel aging patients;

[0027] A result output unit for outputting the classification result.

[0028] The third aspect of the present application provides a computer device.

[0029] Further, the computer device comprises a memory and a processor, the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, the method for evaluating the treatment effect of the anti-aging drug Senolytics according to the first aspect of the present application is realized.

[0030] The fourth aspect of the present application provides a computer readable storage medium.

[0031] Further, the computer readable storage medium stores a computer program, and the computer program is executed by the processor to realize the method for evaluating the treatment effect of the anti-aging drug Senolytics according to the first aspect of the present application.

[0032] The fifth aspect of the present application provides a method for inhibiting vascular endothelial cell aging for non-therapeutic purposes in vitro.

[0033] Further, the method comprises administering a drug combination of dasatinib and quercetin to the vascular endothelial cells.

[0034] Further, the concentration ratio of the dasatinib and quercetin is 1:10.

[0035] The sixth aspect of the present application provides the use of the inhibitor of JPX in the preparation of an anti-aging drug.

[0036] Further, the inhibitor is shRNA.

[0037] Advantages and beneficial effects of the present application:

[0038] The present application first discovers that the drug combination of anti-aging drugs Senolytics dasatinib and quercetin plays an anti-aging role by inhibiting the levels of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex, and based on this, the present application provides a method, system and device for evaluating the therapeutic effect of the drug combination of anti-aging drugs Senolytics dasatinib and quercetin based on the expression level of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex, which has a broad application prospect in the evaluation of the therapeutic effect of cardiovascular disease drugs. BRIEF DESCRIPTION OF DRAWINGS

[0039] Figure 1 A flowchart of a method for evaluating the therapeutic effect of anti-aging drugs Senolytics according to an embodiment of the present application is shown in the figure;

[0040] Figure 2 A schematic diagram of a system for evaluating the therapeutic effect of anti-aging drugs Senolytics according to an embodiment of the present application is shown in the figure;

[0041] Figure 3 A schematic diagram of the structure of a computer device according to the present application is shown in the figure;

[0042] Figure 4 A result graph of anti-aging drugs Senolytics improving LPS-induced HUVECs aging is shown in the figure, wherein, Figure 4 A is a result graph of CCK-8 experiment for detecting the cell viability of HUVECs treated with different concentrations of Das and Que; Figure 4 B is a result graph of CCK-8 experiment for detecting the cell viability of HUVECs treated with different concentrations of Das and Que after LPS intervention; Figure 4 C is a result graph of SA-β-gal staining after treatment with Das and Que; Figure 4 D is BrdU staining after treatment with Das and Que;

[0043] Figure 5 The influence of Senolytics on the expression of aging-related proteins and SASP genes is shown in the figure, wherein, Figure 5A is the relative telomere length result chart; Figure 5 B is the RT-PCR analysis of the effect of Senolytics on the expression level of JPX result chart; Figure 5 C- Figure 5 D is the Western blot analysis of the effect of Senolytics on the expression of senescence-related proteins (p53, p21 and p16); Figure 5 E is the RT-PCR analysis of the effect of Senolytics on the expression of SASP genes (IL-6, IL-8, IL-1β, CCL2, ICAM-1 and TNF-α); "*" indicates compared with the control group, "##" indicates compared with the LPS+Senolytics group;

[0044] Figure 6 is the result chart of JPX gene knockout can alleviate vascular endothelial cell senescence; wherein, Figure 6 A is the RT-PCR detection shows the expression level of JPX; Figure 6 B is the relative telomere length result chart; Figure 6 C is the SA-β-gal staining result chart after JPX gene knockout; Figure 6 D is the BrdU staining result after JPX gene knockout; "*" indicates compared with the control group, "##" indicates compared with the LPS+shNC group;

[0045] Figure 7 is the result chart of the effect of JPX knockout on the expression level of senescence-related proteins and SASP genes in LPS-induced endothelial cells; wherein, Figure 7 A is the Western blot analysis of the effect of JPX knockout on the expression level of senescence-related proteins (p53, p21 and p16); Figure 7 B is the RT-PCR analysis of the effect of JPX knockout on the expression level of SASP genes (IL-6, IL-8, IL-1β, CCL2, ICAM-1 and TNF-α); "*" indicates compared with the control group, "##" indicates compared with the LPS+shNC group;

[0046] Figure 8 is the related result chart of anti-aging drug Senolytics improving vascular endothelial senescence by antagonizing JPX-BRD4-p65 complex; wherein, Figure 8 A is the Western blot analysis of the effect of anti-aging drug Senolytics on BRD4 and NF-κB signaling pathway; Figure 8 B is the RNA-RNA pull down experiment result of detecting the interaction between JPX and BRD4 and p65; Figure 8 C is the RNA immunoprecipitation (RIP) detection to evaluate the association between JPX and BRD4 and p65 proteins;Figure 8 D represents the predicted RNA-protein interaction result. A probability greater than 0.5 indicates a positive result, suggesting that there may be an interaction between RNA and protein. Figure 8 E- Figure 8 F, Chromatin immunoprecipitation (ChIP) showed that p65 and BRD4 were enriched at the SASP gene promoter; "*" indicates comparison with the control group, "##" indicates comparison with the LPS group;

[0047] Figure 9 This figure shows the in vivo activity of senolytics against vascular endothelial cell senescence; among which, Figure 9 A represents Western blot analysis to assess the effects of senolytics on aging-related proteins (p21, p16), BRD4, and the p-p65 / p65 ratio in mouse aortic tissue. Figure 9 B represents the effect of RT-PCR on the expression levels of SASP genes (IL-6, IL-8, IL-1β, CCL2, ICAM-1, and TNF-α) in mouse aortic tissue; "*" indicates the difference compared to the control group, and "##" indicates the difference compared to the normal aging group.

[0048] Figure 10 This image shows the results of the anti-aging drug Senolytics improving vascular endothelial aging in vivo by antagonizing the JPX-BRD4-p65 complex; [The image shows...] Figure 10 A represents the expression levels and co-localization of BRD4 and p65 in mouse aortic vascular tissue analyzed by immunofluorescence. Figure 10 B is a RIP assay to evaluate the effect of senolytics on the formation of the JPX-BRD4-p65 complex; Figure 10 C represents the effect of ChIP assay on the enrichment of the JPX-p65-BRD4 complex on the SASP gene promoter. Detailed Implementation

[0049] To overcome the shortcomings of existing technologies, the overall approach of the technical solution provided in this application is as follows: Through LPS-induced human umbilical vein endothelial cell senescence model and natural senescence model, we demonstrated the anti-aging effect of the drug combination of dasatinib and quercetin; subsequently, through analysis, we found that the drug combination of dasatinib and quercetin exerts its anti-aging effect by inhibiting the formation of the JPX-BRD4-p65 complex; based on this, we propose a method, system, and device for evaluating the therapeutic effect of the anti-aging drug Senolytics.

[0050] To enable those skilled in the art to better understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings.

[0051] In some processes described in the specification, claims, and accompanying drawings of this invention, multiple operations are included in a specific order. However, it should be clearly understood that these operations may not be performed in the order they appear herein, or may be performed in parallel. The operation numbers, such as 101, 102, etc., are merely used to distinguish different operations and do not represent any execution order. Furthermore, these processes may include more or fewer operations, and these operations may be performed sequentially or in parallel.

[0052] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Experimental methods in the following embodiments that do not specify specific conditions are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer.

[0053] Figure 1 This invention provides a flowchart illustrating a method for evaluating the therapeutic effect of the anti-aging drug Senolytics. Specifically, the method includes the following steps:

[0054] 101: Obtain data to obtain expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex in vascular aging patient samples after treatment with the anti-aging drug Senolytics;

[0055] In some embodiments, the patient may be human or non-human and may include, for example, animal strains or species used as a “model system” for research purposes. Similarly, the patient may include adults or adolescents (e.g., children). Furthermore, the patient may refer to any living organism, preferably a mammal (e.g., human or non-human), that may benefit from administration of the pharmaceutical composition of the anti-aging drugs Senolytics dasatinib and quercetin. Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates (e.g., chimpanzees) and other apes and monkeys; livestock, such as cattle, horses, sheep, goats, and pigs; domestic animals, such as rabbits, dogs, and cats; and laboratory animals, including rodents such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds, fish, etc.

[0056] In the context of this invention, the term "sample" as used refers to a composition obtained from or derived from a patient / subject that contains cells and / or other molecular entities to be characterized and / or identified based on, for example, physical, biochemical, chemical, and / or physiological characteristics. For example, a sample refers to any sample derived from a patient / subject that is expected or known to contain cells and / or molecular entities to be characterized. Samples include, but are not limited to, tissue samples, primary or cultured cells or cell lines, cell cultures, cell supernatants, cell lysates, platelets, serum, plasma, vitreous fluid, lymph, synovial fluid, follicular fluid, semen, pancreatic juice, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, tissue culture fluid, tissue extracts, homogenized tissue, cell extracts, and combinations thereof. In a specific embodiment of this invention, the sample is derived from vascular endothelial cells or vascular tissue.

[0057] In this invention, "anti-aging drugs Senolytics", "drug combination of dasatinib and quercetin" and "D+Q" are synonymous and all refer to anti-aging drugs composed of dasatinib and quercetin.

[0058] In this invention, "p-p65 / p65" refers to the proportion of phosphorylated p65 to total p65.

[0059] In some implementations, expression level data for JPX, BRD4, p-p65 / p65, and / or the JPX-BRD4-p65 complex can be detected using methods well-known in the art. For example, expression level data for JPX, BRD4, p-p65 / p65, and / or the JPX-BRD4-p65 complex can be obtained at the nucleic acid level by measuring the amount of RNA, mRNA, or any other RNA species using methods well-known in the art, including digital PCR and real-time (RT) quantitative or semi-quantitative PCR, fluorescence-activated cell sorting (FACS), and in situ hybridization.

[0060] In other embodiments, expression level data for JPX, BRD4, p-p65 / p65, and / or the JPX-BRD4-p65 complex can also be obtained by measuring protein-level expression levels, including mass spectrometry-based quantitative proteomics, immunoassays, Western blotting, spectrophotometry, enzymatic assays, ultraviolet assays, kinetic assays, electrochemical assays, colorimetric assays, turbidimetric assays, atomic absorption spectrometry, flow cytometry, mass flow cytometry, or any combination thereof. In specific embodiments of the invention, Western blotting or RT-PCR is used to detect the expression levels of JPX, BRD4, and p-p65 / p65, and RIP is used to detect the formation of the JPX-BRD4-p65 complex.

[0061] In one embodiment of the present invention, to investigate the protective effect of senolytics, a combination of dasatinib and quercetin (D+Q), on senescent endothelial cells, we established a human umbilical vein endothelial cell (HUVECs) senescence model. HUVECs were treated with 1 μg / mL lipopolysaccharide (LPS) for 24 hours, followed by intervention with senolytics. Initially, to determine the safe and effective concentration of D+Q in HUVECs, cells were treated with different concentrations of dasatinib (Das) and quercetin (Que), and cell viability was assessed using a CCK-8 assay. The results showed that when Das was below 1 μM and Que was below 10 μM, cytotoxicity was not significant. Figure 4 A). Subsequently, LPS-treated HUVECs were exposed to different concentrations of Das and Que. CCK-8 assay results showed that 0.1 μM Das and 1 μM Que significantly reversed the LPS-induced senescence phenotype of HUVECs. Figure 4 B). Therefore, 0.1 μM Das and 1 μM Que were selected as the optimal concentrations for antagonizing LPS-induced endothelial senescence.

[0062] In one embodiment of the invention, to investigate the effect of senolytics on SA-β-gal activity, we performed SA-β-gal staining. For example... Figure 4 As shown in Figure C, compared with the control group, the number of SA-β-gal positive cells in the LPS treatment group was significantly increased, while the intervention of senolytics significantly reduced the number of SA-β-gal positive cells. Furthermore, cell proliferation was assessed by BrdU staining. The results showed that compared with the control group, the number of BrdU positive cells in the LPS group was reduced, indicating impaired proliferation; while the intervention of senolytics significantly increased the number of BrdU positive cells (…). Figure 4 (D) This indicates that senolytics intervention restored the proliferative capacity of senescent cells. Furthermore, relative telomere length analysis showed that LPS significantly shortened telomere length compared to the control group, while senolytics treatment greatly reversed the shortening of telomere length. Figure 5 A). Furthermore, Western blotting and RT-PCR were used to assess the effects of senolytics on the expression of aging-related proteins (p53, p21, and p16) and SASP factors (IL-6, IL-8, IL-1β, CCL2, ICAM-1, and TNF-α). The results showed that, compared to the control group, aging markers and SASP genes were significantly upregulated in the LPS group, while senolytics treatment significantly downregulated their expression levels. Figure 5CE). Notably, RT-PCR analysis also showed that LPS significantly increased JPX expression, while JPX expression significantly decreased after senolytics intervention. Figure 5 (B) This indicates that JPX plays a crucial role in endothelial cell senescence. In summary, these results demonstrate that senolytics can effectively antagonize LPS-induced HUVEC senescence and inhibit JPX expression.

[0063] In one embodiment of the present invention, to further investigate the role of JPX in senescent endothelial cells, we constructed a lentiviral vector encoding a shRNA targeting JPX (shJPX) and transfected it into HUVECs to knock out JPX. RT-PCR analysis showed that after transfection with shJPX, the expression of JPX in LPS-induced senescent myocardium was significantly reduced. Figure 6 A) confirmed the successful knockout of JPX. Subsequently, we evaluated the effects of JPX knockout on telomere length, SA-β-gal activity, cell proliferation, aging-related protein levels, and SASP gene expression in senescent cardiac ECGs. The results showed that JPX knockout significantly prolonged telomere length in LPS-induced senescent ECGs (A). Figure 6 B), reducing the proportion of SA-β-gal positive cells ( Figure 6 C), increases BrdU-positive cells ( Figure 6 D). Furthermore, JPX silencing significantly downregulated the expression of aging markers p53, p21, and p16, and significantly inhibited multiple SASP factors, including IL-6, IL-8, IL-1β, CCL2, ICAM-1, and TNF-α. Figure 7 A- Figure 7 B). In summary, these findings suggest that JPX promotes endothelial cell senescence, while senolytics can delay vascular endothelial senescence by inhibiting JPX expression.

[0064] In one embodiment of the present invention, to investigate whether senolytics exerts an anti-aging effect by regulating the expression of p65 and BRD4, we first performed Western blot analysis. The results showed that LPS treatment significantly increased the ratio of phosphorylated p65 to total p65 (p-p65 / p65) and elevated BRD4 protein levels, while senolytics treatment effectively inhibited these increases. This suggests that senolytics may combat aging by regulating the BRD4 and NF-κB signaling pathways. Figure 8 A). Furthermore, to verify whether JPX directly interacts with p65 and BRD4, we used the RPISeq computational tool to predict the binding potential of JPX with these two proteins. The analysis results indicate that JPX is highly likely to interact with BRD4 and p65.Figure 8 D). Subsequent RNA pull-down and RNA immunoprecipitation (RIP) experiments showed that JPX physically binds to BRD4 and p65, forming a JPX-BRD4-p65 complex within the cell. Figure 8 BC). Finally, the functional roles of p65 and BRD4 in regulating SASP-related genes were investigated using chromatin immunoprecipitation (ChIP) experiments. Figure 8 Data showed that LPS stimulation significantly enhanced the binding of p65 and BRD4 to the promoter regions of SASP genes, including IL-6, IL-8, CCL2, and ICAM-1, indicating that they play a key role in the transcriptional activation of these genes. These findings suggest that senolytics exert their anti-aging effects by antagonizing the JPX-BRD4-p65 complex.

[0065] In one embodiment of the present invention, to further investigate the anti-aging effects and underlying mechanisms of senolytics in vivo, a naturally aging mouse model was established using 20-month-old C57BL / 6 mice and 8-week-old C57BL / 6 mice as controls. Western blot analysis showed that, compared with the control group, the expression levels of aging-related proteins (p21, p16), BRD4, and the ratio of phosphorylated p65 to total p65 (p-p65 / p65) in the aortic tissue of naturally aging mice were significantly upregulated. Figure 9 A). Senolytics intervention significantly reduced the expression of these proteins, indicating its efficacy in alleviating vascular endothelial aging in vivo. Further RT-PCR analysis showed that the expression of SASP-related genes (IL-6, IL-8, IL-1β, CCL2, ICAM-1, and TNF-α) was significantly increased in the naturally aging group, while senolytics significantly inhibited the expression of these inflammatory genes. Figure 9 B), indicating that it can effectively alleviate age-related inflammatory phenotypes. To explore the potential molecular mechanisms, researchers performed immunofluorescence staining to assess the expression levels and co-localization of BRD4 and p65 in aortic tissue. Figure 10 A). The results showed that the expression and co-localization of BRD4 and p65 were increased in the naturally aging group, while this phenomenon was greatly reversed after senolytics treatment, suggesting that senolytics may exert its effects by inhibiting the BRD4 and NF-κB signaling pathways. Subsequent RIP assays showed that the interaction between BRD4-p65 and JPX was significantly enhanced in the naturally aging group, while it was significantly weakened under the influence of senolytics. Figure 10(B) This indicates that senolytics disrupts the formation of the JPX-BRD4-p65 complex. Consistent with this, ChIP assays showed that in naturally aged mice, the binding of the JPX-BRD4-p65 complex to the promoter regions of SASP genes (IL-6, IL-8, CCL2, and ICAM-1) was significantly increased, while it was significantly reduced after senolytics treatment. This suggests that senolytics inhibits SASP gene expression by interfering with complex formation. Figure 10 C). In summary, these findings demonstrate that senolytics effectively alleviate vascular endothelial aging in vivo by inhibiting the formation of the JPX-BRD4-p65 complex and the resulting SASP gene expression, which is consistent with our in vitro results.

[0066] 102: Process the data by inputting the expression data of JPX, BRD4, p-p65 / p65 and / or the JPX-BRD4-p65 complex into the constructed evaluation model, which predicts the therapeutic effect of the anti-aging drug Senolytics based on the expression data of JPX, BRD4, p-p65 / p65 and / or the JPX-BRD4-p65 complex.

[0067] In some implementations, the methods for constructing the evaluation model are known to those skilled in the art and can be implemented and realized in different ways, linking the expression levels of JPX, BRD4, p-p65 / p65 and / or the JPX-BRD4-p65 complex with a certain probability or risk.

[0068] In the context of this invention, the term "machine learning" refers to the use of computers to simulate or implement human learning activities, and technicians typically use various development tools to build machine learning algorithmic models. These development tools include, but are not limited to, TensorFlow, Scikit-Learn, PyTorch, OpenNN, RapidMiner, Azure Machine Learning, Apache Mahout, Shogun, KNIME, Vertex AI, H2Oai, Anaconda, Keras, Tableau, Fast.ai, Catalyst, Amazon ML, MLJAR, and Spell. The algorithmic models include, but are not limited to, linear regression models, logistic regression models, Lasso regression models, Ridge regression models, linear discriminant analysis models, nearest neighbor models, decision tree models, perceptron models, neural network models, support vector machine models, Naive Bayes models, AdaBoost models, GBDT models, XGBoost models, LightGBM models, CatBoost models, or random forest models.

[0069] In one embodiment, after constructing the evaluation model, the effectiveness of the evaluation model can be analyzed using ROC curves.

[0070] An ROC curve is a graph of the true positive rate (sensitivity) versus the false positive rate (100% specificity) of an experiment. It is useful for depicting the performance of a specific characteristic when distinguishing between two populations. Typically, characteristic data are selected across the entire population in ascending order based on the values ​​of a single characteristic. Then, for each value of that characteristic, the true positive and false positive rates of the data are calculated. The true positive rate is determined by counting the number of cases with values ​​higher than the characteristic value and dividing by the total number of cases. The false positive rate is determined by counting the number of controls with values ​​higher than the characteristic value and dividing by the total number of controls. While this definition refers to cases where the characteristic is higher in cases compared to controls, it also applies to cases where the characteristic is lower in cases compared to controls (in which case samples with values ​​lower than the characteristic value are counted). ROC curves can be generated with respect to individual characteristics and can also be generated with respect to other individual outputs. For example, combinations of two or more characteristics can be mathematically combined (e.g., addition, subtraction, multiplication, etc.) to provide individual sum values ​​that can be plotted on the ROC curve. Furthermore, any combination of multiple features derived from individual output values ​​can be plotted on a ROC curve.

[0071] 103: Output results.

[0072] In this invention, the evaluation model obtains results using the following criteria: when the expression levels of JPX, BRD4, p-p65 / p65, and / or the JPX-BRD4-p65 complex are below a threshold, a classification result indicating that the anti-aging drug Senolytics is effective in treating patients with vascular aging is obtained; when the expression levels of JPX, BRD4, p-p65 / p65, and / or the JPX-BRD4-p65 complex are above a threshold, a classification result indicating that the anti-aging drug Senolytics is ineffective in treating patients with vascular aging is obtained.

[0073] In some embodiments, the preset threshold is a representative value of a normal sample from a vascular aging population, including but not limited to the maximum value, the third or fourth quartile, and the mean. In some preferred embodiments of the present invention, the population sample includes 20 or more samples, such as 30, 50, 80, 100, 150, 200, 300, 500, or more.

[0074] Figure 2 This is a schematic diagram of a system for evaluating the therapeutic effect of the anti-aging drug Senolytics, provided as an embodiment of the present invention.

[0075] The system is programmed or otherwise configured to include a data acquisition unit 201, a data classification unit 202, and a result output unit 203.

[0076] Data acquisition unit 201: used to acquire expression data of JPX, BRD4, p-p65 / p65 and / or JPX-BRD4-p65 complex in vascular aging patient samples after treatment with the anti-aging drug Senolytics;

[0077] Data classification unit 202: used to classify and predict the data obtained by the data acquisition unit through the evaluation model obtained by the construction method described in the first aspect of the present invention, and to obtain the classification result of whether the anti-aging drug Senolytics is effective in treating patients with vascular aging.

[0078] Result output unit 203: Used to output classification results.

[0079] The system may be a user's electronic device or a computer system remotely located relative to that electronic device.

[0080] Figure 3 A schematic diagram of the structure of the computer device provided by the present invention.

[0081] The computer device 300 includes a processor 301 and a memory 302 coupled to the processor 301. The memory 302 stores program instructions. When the program instructions are executed by the processor 301, the processor 301 performs the method described above for achieving the therapeutic effect of the anti-aging drug Senolytics.

[0082] The processor 301 can also be referred to as a CPU (Central Processing Unit). The processor 301 may be an integrated circuit chip with signal processing capabilities. The processor 301 can also be a general-purpose processor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA), or other programmable logic devices, discrete gate or transistor logic devices, or discrete hardware components. A general-purpose processor can be a microprocessor or any conventional processor.

[0083] Computer device 300 can be a mobile electronic device.

[0084] It should be understood that the systems, apparatuses, and methods described in this invention can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative; for instance, the division of modules is only a logical functional division, and in actual implementation, there may be other division methods. For example, multiple modules or components may be combined or integrated into another system, or some features may be ignored or not executed. Furthermore, the couplings or direct couplings or communication connections shown or discussed may be indirect couplings or communication connections through some interfaces, apparatuses, or modules, and may be electrical, mechanical, or other forms.

[0085] The modules described as separate components may or may not be physically separate. The components shown as modules may or may not be physical units; that is, they may be located in one place or distributed across multiple network units. Some or all of the units can be selected to achieve the purpose of this embodiment according to actual needs.

[0086] Furthermore, the functional modules in the various embodiments of the present invention can be integrated into one processing module, or each module can exist physically separately, or two or more modules can be integrated into one module. The integrated modules described above can be implemented in hardware or as software functional modules.

[0087] The above are merely embodiments of this application and do not limit the scope of this patent application. Any equivalent structural or procedural changes made using the content of this application's specification and drawings, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of this application.

Claims

1. A method of evaluating the therapeutic effect of the anti-aging drug Senolytics, characterized by, The anti-aging drug Senolytics is a drug combination of dasatinib and quercetin, the method is completed by a computer, and the method comprises the following steps: Data acquisition: the expression data of JPX, BRD4, p65 / p65 and / or JPX-BRD4-p65 complex in the samples of vascular aging patients after treatment with anti-aging drug Senolytics are acquired, p -p65 / p65 and / or JPX-BRD4-p65 complex; processing data: inputting the expression data of the JPX, BRD4, p -p65 / p65 and / or JPX-BRD4-p65 complex into the evaluation model constructed based on the expression data of the JPX, BRD4, p -p65 / p65 and / or JPX-BRD4-p65 complex predicts the therapeutic effect of the anti-aging drug Senolytics; outputting the result.

2. The method of claim 1, wherein, The construction steps of the evaluation model are as follows: Get JPX, BRD4, p Expression data of p65 / p65 and / or JPX-BRD4-p65 complexes, wherein JPX, BRD4, p Expression data for p65 / p65 and / or the JPX-BRD4-p65 complex were obtained from untreated patients with vascular aging and patients with vascular aging treated with the anti-aging drug Senolytics. JPX, BRD4, p - Expression data of p65 / p65 and / or JPX-BRD4-p65 complex is inputted into machine learning algorithm to build an evaluation model.

3. The method of claim 2, wherein, The machine learning algorithm comprises an algorithm model developed by using various development tools; Preferably, the development tools comprise but are not limited to TensorFlow, Scikit Learn, PyTorch, OpenNN, RapidMiner, Azure Machine Learning, Apache Mahout, Shogun, KNIME, Vertex AI, H2Oai, Anaconda, Keras, Tableau, Fast.ai, Catalyst, Amazon ML, MLJAR, and Spell. Preferably, the algorithm model comprises but is not limited to a linear regression model, a logistic regression model, a Lasso regression model, a Ridge regression model, a linear discriminant analysis model, a neighbor model, a decision tree model, a perception model, a neural network model, a support vector machine model, a naive Bayes model, an AdaBoost model, a GBDT model, an XGBoost model, a LightGBM model, a CatBoost model, and a random forest model.

4. The method of claim 1, wherein, The evaluation model obtains the result through the following standards: when the expression amount of the JPX, BRD4, p when the expression amount of the JPX, BRD4, p when the expression amount of the JPX, BRD4, 5. A system for evaluating the effectiveness of treatment with anti-aging drugs Senolytics, characterized by, The anti-aging drug Senolytics is a drug combination of dasatinib and quercetin, and the system comprises: Data acquisition unit: for acquiring the expression data of JPX, BRD4, p65 / p65 and / or JPX-BRD4-p65 complex in the blood vessel aging patient sample after the treatment of anti-aging drug Senolytics, p -p65 / p65 and / or JPX-BRD4-p65 complex a data classification unit: used for classifying and predicting the data obtained in the data acquisition unit by the evaluation model obtained by the construction method of claim 2, to obtain a classification result of whether the anti-aging drug Senolytics is effective for treating vascular aging patients; a result output unit: used for outputting the classification result.

6. A computer device, comprising: The computer device comprises a memory and a processor, the memory is used for storing program instructions; and the processor is used for calling the program instructions, and when the program instructions are executed, the method for evaluating the treatment effect of the anti-aging drug Senolytics according to any one of claims 1-4 is realized.

7. A computer readable storage medium characterized in that, The computer readable storage medium stores a computer program, and the computer program is executed by the processor to realize the method for evaluating the treatment effect of the anti-aging drug Senolytics according to any one of claims 1-4.

8. A method of inhibiting senescence of vascular endothelial cells in vitro for non-therapeutic purposes, characterized in that, The method comprises administering the drug combination of dasatinib and quercetin to vascular endothelial cells.

9. The method of claim 8, wherein, The concentration ratio of the dasatinib and quercetin is 1:

10.

10. Use of an inhibitor of JPK in the preparation of an anti-aging drug; Preferably, the inhibitor is shRNA.

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