Application of rhynchophylline in preparation of medicine for treating hypertensive vascular endothelial injury

Abstract

The invention relates to application of rhynchophylline in preparation of a medicine for treating hypertensive vascular endothelial injury, and belongs to the technical field of biology. According to the invention, the treatment effect of the rhynchophylline on hypertension vascular endothelial injury and EPCs dysfunction is defined; researches find that rhynchophylline can be used for preparing an SIRT3 activator, and rhynchophylline can activate SIRT3 in a targeted manner and inhibit a downstream mitochondrial apoptosis pathway, so that EPCs mitochondrial injury is improved, EPCs dysfunction is antagonized, endothelial injury repair is promoted, and the effects of regulating blood pressure, improving the endothelial function and inhibiting hypertension target organ injury are achieved. Good application prospects are realized in medicines for clinically treating hypertension.

Description

Technical Field

[0001] The invention relates to application of rhynchophylline in preparing medicine for treating hypertension vascular endothelial injury, belonging to the field of biotechnology. Background Art

[0002] The information disclosed in this background technology section is only intended to increase some understanding of the overall background of the invention, and should not be necessarily regarded as an admission or any form of suggestion that the information constitutes the prior art already known to ordinary technicians in this field.

[0003] Essential hypertension, also known as hypertension, is a progressive cardiovascular disease characterized by sustained elevation of arterial blood pressure. Hypertension is the most common chronic disease in clinical practice and the most important risk factor for cardiovascular and cerebrovascular diseases. It is often accompanied by disorders of fat and sugar metabolism and functional or organic changes in organs such as the heart, brain, kidney and retina. Major complications include stroke, myocardial infarction, heart failure and chronic kidney disease. The pathological mechanism of hypertension is complex. In recent years, a large number of studies have shown that hypertension is closely related to endothelial damage and endothelial dysfunction. Under the regulation of cytoskeletal elements and cell adhesion complexes, vascular endothelial cells constitute a semi-selective barrier for macromolecules and cellular elements. Studies have found that the endothelium plays a vital role in maintaining the stability of the vascular environment. Endothelial dysfunction can cause an increase in systemic vascular resistance, leading to hypertension. Increased blood pressure can further aggravate endothelial damage and accelerate the progression of hypertension. Therefore, endothelial dysfunction is considered to be an early event in the pathological process of hypertension and a predisposing factor for target organ damage. Repairing endothelial damage and improving endothelial function are of great significance for blood pressure regulation and target organ protection.

[0004] Endothelial progenitor cells (EPCs) are immature precursor cells of endothelial cells. They are a type of directed stem cells that have the characteristics of migration between stem cells and vascular endothelial cells and can self-renew, proliferate and differentiate. They are also called angioblasts. EPCs originate from the bone marrow and circulate in the peripheral blood. They promote the maturation and differentiation of endothelial cells and promote angiogenesis by producing and secreting a variety of angiogenic growth factors and cytokines. Therefore, EPCs can repair damaged endothelium and blood vessels and are considered to be the most important endogenous vascular repair pathway. However, recent studies have shown that age and cardiovascular risk factors can reduce the bioavailability of circulating EPCs and impair their function to varying degrees. Hypertension is an important independent risk factor for impaired EPC function. Studies have found that patients with hypertension have reduced EPC numbers, decreased cell function, and reduced endothelial repair capacity. In addition, various pathological mechanisms in the course of hypertension, such as abnormal autonomic nervous system, abnormal renin-angiotensin-aldosterone system, and inflammation, can lead to EPC dysfunction. Therefore, how to improve the reduction in the number and abnormal function of EPCs in hypertensive patients is of great significance to endothelial repair and blood pressure regulation, and has become a new approach for the treatment of hypertension.

[0005] Mitochondria are involved in cellular energy metabolism, information transmission, cell differentiation, apoptosis and other processes. Abnormal mitochondrial structure and function affect the overall function of cells, leading to a variety of diseases. Recent studies have shown that regulating mitochondrial fission and apoptosis can repair dysfunctional EPCs and reduce oxidative stress damage to EPCs, which may provide new ideas for the protection of hypertensive EPCs. Other studies have found that histone deacetylase SIRT3, as a key regulator of mitochondrial oxidation and metabolic function, is involved in a variety of oxidative stress and aging-related diseases, such as atherosclerosis, myocardial hypertrophy, and hypertension. The study found that the overall loss of SIRT3 increased the blood pressure of model mice, aggravated vascular aging and inflammation, while overexpression of SIRT3 improved endothelial function and inhibited blood pressure increases, suggesting the therapeutic potential of this mitochondrial target for hypertension and endothelial damage.

[0006] Seeking antihypertensive drugs with good target organ protection has always been the goal of clinicians and scientific researchers. With the deepening of the understanding of the cardiovascular protective effect of EPCs, researchers have gradually regarded EPCs as an effective target for the treatment of hypertension. Based on the holistic concept and the theory of syndrome differentiation and treatment, traditional Chinese medicine for the treatment of hypertension has significant advantages in target organ protection on the basis of effective blood pressure reduction. However, the complex effective ingredients of traditional Chinese medicine prescriptions, unclear mechanisms of action, and lack of targeted therapeutic drugs have always plagued the modernization of traditional Chinese medicine and affected its clinical application and promotion. Therefore, further clarifying the efficacy and target of rhynchophylline, the active ingredient of the clinically effective prescription (Tengfu Antihypertensive Prescription), has important exploration value. At the same time, it is a bold innovation and application to promote the integration of traditional Chinese medicine treatment with international hypertension research and accelerate the scientific and internationalization of traditional Chinese medicine. Summary of the invention

[0007] In view of the deficiencies in the prior art, the present invention clarifies the therapeutic effect of Rhynchophylline on hypertensive vascular endothelial damage and EPCs dysfunction; the present invention has found that Rhynchophylline can be used to prepare SIRT3 activators, and Rhynchophylline can improve EPCs mitochondrial damage, antagonize EPCs dysfunction, and promote endothelial damage repair by targeted activation of SIRT3 and inhibition of downstream mitochondrial apoptosis pathways, thereby achieving the effects of regulating blood pressure, improving endothelial function, and inhibiting hypertensive target organ damage, and has good application prospects in clinical drugs for the treatment of hypertension.

[0008] The technical solution adopted by the present invention is as follows:

[0009] In a first aspect of the present invention, a use of rhynchophylline in preparing a histone deacetylase 3 (SIRT3) activator is provided.

[0010] Rhynchophylline is a white crystalline powder with a CAS number of 76-66-4. Its chemical name is Corynixan-16-carboxylic acid, 16, 17-didehydro-17-methoxy-2-oxo-, methyl ester, (7B, 16E, 20a)-, and its molecular formula is C 22 H 28 N 2 O 4 , molecular weight is 384.47 g / mol, soluble in chloroform, acetone, ethanol, benzene, chemical structure is Figure 1 shown.

[0011] Experiments have shown that rhynchophylline can act as a SIRT3 activator to enhance SIRT3 expression in EPCs.

[0012] In a second aspect of the present invention, a use of a SIRT3 activator in the preparation of a drug for treating hypertension vascular endothelial injury is provided.

[0013] In a third aspect of the present invention, there is provided a use of rhynchophylline in the preparation of a medicament for preventing and / or treating hypertensive vascular endothelial damage.

[0014] Experiments have shown that rhynchophylline can maintain the morphology and structural integrity of the vascular endothelium, regulate vascular contraction and relaxation factors, and improve endothelium-dependent vasodilation function; it can inhibit vascular fibrosis by inhibiting collagen deposition, delay the process of vascular fibrosis, and thus reduce vascular wall damage. Rhynchophylline enhances EPCs mitochondrial respiration, significantly increases cell oxygen consumption rate and ATP generation rate; it can also significantly increase EPCs mitochondrial membrane potential.

[0015] The optimal dosage of rhynchophylline for the treatment of spontaneously hypertensive rats (SHR) model was 0.862 mg kg -1 ·d -1 The optimal dosage for treating EPCs dysfunction model in vitro was 100 μmol·L -1 .

[0016] Compared with the related art known to the inventor, one of the technical solutions of the present invention has the following beneficial effects:

[0017] (1) The present invention provides the use of rhynchophylline in the preparation of a drug for treating hypertension vascular endothelial damage;

[0018] (2) The present invention designs and implements pharmacological experiments, and verifies for the first time that Rhynchophylline can be used as a SIRT3 activator, which inhibits the downstream mitochondrial apoptosis pathway by activating SIRT3, thereby antagonizing mitochondrial damage in endothelial progenitor cells (EPCs), alleviating EPCs dysfunction, and promoting endothelial damage repair, thereby achieving the effects of regulating blood pressure and antagonizing target organ damage, providing a new approach to broaden the screening and development of SIRT3 activators and antihypertensive drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their description are used to explain the present invention and do not constitute improper limitations on the present invention.

[0020] Figure 1 Rhynchophylline chemical structure.

[0021] Figure 2 Rhynchophylline significantly reduced blood pressure in SHR rats.

[0022] Figure 3 Rhynchophylline has a protective effect on the vascular endothelium of SHR rats.

[0023] Figure 4 Rhynchophylline improves the function of EPCs in SHR rats.

[0024] Figure 5 Rhynchophylline can act as a SIRT3 activator to enhance SIRT3 expression in EPCs.

[0025] Figure 6 Knockout of SIRT3 can reverse the effect of rhynchophylline on improving mitochondrial dysfunction in EPCs.

[0026] Figure 7 Knockout of SIRT3 can reverse the effect of rhynchophylline on antagonizing EPCs injury. DETAILED DESCRIPTION

[0027] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0028] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, it indicates the presence of features, steps, operations and / or combinations thereof.

[0029] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0030] Example 1: Effects of Rhynchophylline on Endothelial Damage and EPCs Function in SHR Rats

[0031] 1. Experimental Animals

[0032] Sixteen 8-week-old male SHR rats and eight 8-week-old male WKY rats were selected and fed with normal diet. All experimental animals were caged, 4 per cage, with free access to water, and kept in an environment with 12 hours of light and 12 hours of dark.

[0033] 2. Rhynchophylline ingredients and their quality control

[0034] According to the results of previous studies, the optimal intervention concentration of rhynchophylline was 0.862 mg kg -1 To study the factors, HPLC was used to determine the content and perform quality control.

[0035] 3. Experimental Animal Grouping and Drug Administration

[0036] SHR rats were randomly divided into 2 groups, 8 in each group, and 8 WKY rats were used as the normal control group. The rats in each group were adaptively fed for 1 week before drug intervention. The drug was administered at regular intervals every day, 6 days a week, and for 6 consecutive weeks. The specific grouping and intervention measures are as follows:

[0037] Normal control group: WKY rats were intragastrically administered with an equal volume of saline daily;

[0038] Disease model group: SHR rats were intragastrically administered with an equal volume of normal saline daily;

[0039] Rhy intervention group: SHR rats were given Rhy 0.862 mg kg daily by oral gavage -1 .

[0040] 4. Blood pressure monitoring

[0041] The non-invasive tail cuff method was used to detect the tail artery blood pressure of rats. The rat was placed in the fixture of the platform, and the tail cuff was passed through the tail of the rat and as close to the base of the tail as possible. Preheat to 37°C for about 10 minutes to allow the rat to adapt to the stimulation of the platform and keep the rat in a quiet state. Turn on the animal non-invasive blood pressure analysis system, first perform 5 preliminary measurements to promote good blood flow in the tail of the rat to ensure the stability and reliability of the measured value, then measure 3 times in a row, and take the average value as the measured blood pressure value (unit: mmHg). Measure once at the same time every week and monitor continuously for 6 weeks.

[0042] 5. Specimen Collection

[0043] After the intervention, the rats were fasted for 12 h and allowed to drink water freely. Blood was collected from the medial canthal venous plexus of the orbit and placed in a blood collection tube containing a cell separation anticoagulant for the separation and extraction of EPCs from peripheral blood. 1% sodium pentobarbital (50 mg kg -1 ) After intraperitoneal injection of anesthesia, the abdominal cavity was quickly opened and the inferior vena cava blood was collected using a non-anticoagulant tube. The tube was allowed to stand at room temperature for 30 min and centrifuged at 3500 rpm for 15 min. The supernatant was dispensed into 1.5 mL EP tubes and stored in a -80°C refrigerator for later use.

[0044] After blood collection, the chest cavity was opened quickly to fully expose the heart and aorta, and the rat aorta was stripped to the bifurcation of the common iliac artery. The entire aorta was removed together with the heart, and the blood vessels were separated from the heart at the root of the aorta. A part of the blood vessels was cut and placed in 4% paraformaldehyde fixative for the preparation of paraffin sections. The thoracic aorta segment was then cut and immersed in fresh Krebs solution. The residual fat, connective tissue and small branches on the blood vessels were removed under a dissecting microscope, and a 3-4 mm long vascular ring was prepared. The in vitro vascular ring tension test was immediately performed. The remaining aorta was placed in a -80°C refrigerator for future use.

[0045] 6. Isolation and Culture of EPCs from Rat Peripheral Blood

[0046] Take fresh anticoagulated blood, centrifuge at 250g for 10min, discard the plasma, add FBS with the same volume as the discarded plasma and resuspend. Add separation solution 1, separation solution 2 and cell suspension to the centrifuge tube in turn, and centrifuge at 500g for 30min. Carefully pipette the second layer of ring-shaped milky white EPCs layer into another centrifuge tube and wash 3 times. Resuspend the cells in the culture medium, inoculate them in a 3cm culture dish coated with FN, and place them in a cell culture incubator for culture. After 48h, change the medium to remove non-adherent cells, and add 3mL EGM-2MV culture medium to continue culturing. Thereafter, change the culture medium every 2-3 days.

[0047] 7. Index detection and detection methods

[0048] (1) Detection of tail artery systolic and diastolic pressure in awake rats: using the non-invasive tail cuff method;

[0049] (2) Histological morphology of the aorta: HE staining and Masson staining were used to observe the structural changes of the aorta; CD31 immunofluorescence staining was used to observe the integrity of the aortic endothelium.

[0050] (3) Vascular relaxation function testing: in vitro vascular ring tension testing; serum NO and ET-1 content testing.

[0051] (4) Aortic EPCs aggregation detection: Immunofluorescence staining was used to detect the expression of CD133 and CD34 in the aorta;

[0052] (5) Functional testing of peripheral blood EPCs: Transwell assay to detect cell migration ability; adhesion assay to detect cell adhesion ability; Tube formation assay to detect cell tube formation ability.

[0053] 8. Statistical analysis methods

[0054] SPSS statistics 22.0 software was used to analyze the data, and the measurement data were expressed as mean ± standard deviation. If the measurement data among multiple groups were in accordance with normal distribution, one-way analysis of variance (ANOVA) was used, and if they were not in accordance with normal distribution, non-parametric test (Kruskal-Wallis test) was used. P<0.05 indicated that the difference was statistically significant.

[0055] 9. Experimental Results

[0056] Rhynchophylline can maintain the morphology and structural integrity of the vascular endothelium, regulate vasoconstriction and relaxation factors, and improve endothelium-dependent vasodilation function.

[0057] (1) Rhynchophylline significantly reduces blood pressure in SHR rats

[0058] The results showed that the systolic and diastolic blood pressure levels of WKY and SHR rats increased slightly with age, but there was no statistical difference compared with before administration (P>0.05). Compared with WKY rats, the systolic and diastolic blood pressure of SHR rats were significantly increased (P<0.001). After 3 weeks of administration, the systolic and diastolic blood pressure of rats in the SHR+Rhy group decreased significantly, which was statistically different from the SHR group (P<0.001). As the intervention time prolonged, rhynchophylline continued to exert its antihypertensive effect. After 6 weeks of intervention, the blood pressure of rats in the SHR+Rhy group dropped to the lowest, and the difference was statistically significant compared with the SHR group (P<0.001).

[0059] (2) Rhynchophylline has a protective effect on the vascular endothelium of SHR rats

[0060] Figure 3 The results of H&E staining of rat aorta in A and 3D showed that the thoracic aorta of WKY rats had clear wall, thin vascular wall, smooth lumen, intact endothelial structure, normal endothelial cell morphology, uniform size and distribution, and regular cell arrangement; the wall of SHR was rough and hypertrophic, the endothelial cells were degenerated and swollen, the size and distribution were uneven, and the cells were disordered; after the intervention of rhynchophylline, the thickness of the aorta wall of rats was reduced, the lumen was smoother, the vascular endothelial structure tended to be intact, and the size and distribution of endothelial cells were more uniform.

[0061] Figure 3 The results of Masson staining of the aorta of rats B and 3E showed that the area of ​​collagen fibers in the aorta of SHR was significantly increased compared with that of WKY rats (P<0.001). After 6 weeks of rhynchophylline intervention, the area of ​​collagen in the aorta of rats decreased (P<0.001), indicating that rhynchophylline can inhibit vascular fibrosis by inhibiting collagen deposition, delaying the process of vascular fibrosis, and thus reducing vascular wall damage.

[0062] Figure 3The results of immunofluorescence staining of C rat aorta showed that the endothelial marker CD31 was evenly and intactly distributed in the aorta of WKY rats, while the expression level of CD31 in the aorta of SHR rats was reduced, indicating that the endothelial structure was broken and missing. Rhynchophylline can increase the expression of CD31 in the aorta of SHR rats, alleviate endothelial defects, and maintain a relatively complete endothelial structure.

[0063] Figure 3 The results of the isolated vascular ring tension test of F rats showed that compared with WKY, the endothelium-dependent vasodilation activity induced by Ach stimulation in SHR was significantly decreased (P < 0.05). After 6 weeks of rhynchophylline intervention, the sensitivity of rats to Ach stimulation was restored, and the endothelium-dependent vasodilation function was significantly improved, which was significantly different from the SHR group (P < 0.001). The results showed that rhynchophylline can improve the endothelial vasodilation function of hypertensive rats and inhibit endothelial damage in hypertension.

[0064] Figure 3 The results of serum NO content detection in G rats showed that compared with WKY, the serum NO content in SHR was lower, and the difference was statistically significant (P<0.001); while the serum NO level of rats in the rhynchophylline group was significantly higher than that in the SHR group (P<0.01).

[0065] Figure 3 The results of serum ET-1 content detection in H rats showed that compared with WKY, the serum ET-1 content in SHR was significantly increased (P<0.001); the serum ET-1 level in the rhynchophylline group was significantly decreased, which was significantly different from that in the SHR group (P<0.001).

[0066] (3) Rhynchophylline improves EPCs function in SHR rats

[0067] Figure 4 A The results of immunofluorescence double-labeling staining of rat aorta showed that there was no obvious yellow fluorescent area in the aorta of WKY rats, indicating that WKY rats had no endothelial damage, so the number of EPCs gathered in the aortic endothelium was very small. There was a small amount of yellow fluorescent area in the aorta of SHR, and after rhynchophylline intervention, the yellow fluorescent area of ​​the rat aorta increased significantly, indicating that CD133 in the aorta of SHR + / CD34 + The number of labeled EPCs is extremely small, indicating that the migration and adhesion abilities of EPCs in SHR are reduced, and their mobilization and homing to endothelial damage are impaired. Rhynchophylline can improve the function of EPCs, mobilize EPCs to migrate and adhere to damaged vascular endothelium, thereby increasing the number of EPCs in the SHR aorta and promoting the repair of endothelial damage.

[0068] Figure 4B and 4E Transwell experiments showed that compared with the peripheral blood EPCs of WKY rats, the number of migrating cells and the migration ability of peripheral blood EPCs of SHR rats were significantly reduced (P<0.001). The migration ability of peripheral blood EPCs of rats in the rhynchophylline group was improved, and the number of migrating cells was significantly increased compared with that in the SHR group (P<0.05).

[0069] Figure 4 The results of C and 4F adhesion experiments showed that compared with WKY rats, the number of EPCs adhesion cells in the peripheral blood of SHR rats was significantly reduced (P<0.001), indicating that the adhesion ability of EPCs in the peripheral blood of SHR rats was significantly reduced. The adhesion ability of EPCs in the peripheral blood of the rats in the rhynchophylline group was improved, and the number of adhesion cells was significantly increased compared with the model group (P<0.001).

[0070] Figure 4 The results of D and 4G Tube formation experiments showed that compared with WKY rats, the in vitro tube formation ability of EPCs in peripheral blood of SHR rats was reduced, and the length of tubes and the number of branch nodes were significantly reduced, and the difference was statistically significant (P < 0.05); Rhynchophylline intervention can significantly promote the tubular differentiation ability of EPCs in peripheral blood of rats, promote tube formation and connection, and significantly increase the length of tubes and the number of branch nodes, which are significantly different from those of the SHR group (P < 0.05).

[0071] Example 2: Rhynchophylline targets and regulates SIRT3 to improve EPCs mitochondrial function and antagonize EPCs damage

[0072] 1. Culture of Human Bone Marrow-derived EPCs

[0073] Human bone marrow-derived EPCs were selected and placed in a 37°C, 5% CO 2 , cultured in a cell culture incubator with saturated humidity, the culture medium was replaced every 2-3 days, and after the cells grew and merged to 90% of the area of ​​the culture flask, the cells were subcultured at a ratio of 1:2, and the 3rd to 7th generations were used for experiments.

[0074] 2. Rhynchophylline ingredients and their quality control

[0075] The optimal cell administration concentration of rhynchophylline was obtained through CCK8 experiments in the early stage, which was 100 μmol·L -1 The intervention time was 24 h, and the quality control was the same as in Example 1.

[0076] 3. Grouping and intervention

[0077] The cells were randomly assigned to the following groups and the interventions were as follows:

[0078] (1) si-NC group: transfected with NC siRNA for 48 h.

[0079] (2) si-NC+Ang II group: 24 h after transfection with NC siRNA, Ang II (2×10 -6 mol·L -1 ) intervention for 24 hours.

[0080] (3) si-NC+Ang II+Rhy group: 24 h after transfection with NC siRNA, rhynchophylline (100 μmol·L -1 ) for 1 h, and then Ang II (2×10 -6 mol·L -1 ) intervention for 24 hours.

[0081] (4) si-SIRT3 group: transfected with SIRT3 siRNA for 48 h.

[0082] (5) si-SIRT3+Ang II group: 24 h after SIRT3 siRNA transfection, Ang II (2×10 -6 mol·L -1 ) intervention for 24 hours.

[0083] (6) si-SIRT3+Ang II+Rhy group: 24 h after SIRT3 siRNA transfection, rhynchophylline (100 μmol·L -1 ) for 1 h, and then Ang II (2×10 -6 mol·L -1 ) intervention for 24 hours.

[0084] 4. Detection indicators and detection methods

[0085] (1) Detection of SIRT3 expression level: Western blot was used to detect SIRT3 protein expression in EPCs;

[0086] (2) SIRT3 activity detection: SIRT3 activity in EPCs was detected using a SIRT3 activity test kit;

[0087] (3) Mitochondrial respiratory function test: The Seahorse mitochondrial stress test was used to test mitochondrial respiratory function;

[0088] (4) Mitochondrial membrane potential detection: JC-1 fluorescence staining method was used to detect mitochondrial membrane potential;

[0089] (5) Cell function detection: EdU assay to detect cell proliferation ability; Transwell assay to detect cell migration ability; Tube formation assay to detect cell tube formation ability.

[0090] 5. Statistical Analysis

[0091] Same as Example 1.

[0092] 6. Experimental Results

[0093] (1) Rhynchophylline can act as a SIRT3 activator to enhance SIRT3 expression in EPCs

[0094] Figure 5 A Results showed that compared with the normal group, the expression level of SIRT3 protein in EPCs was significantly decreased after Ang II intervention (P<0.001). Rhynchophylline increased SIRT3 protein expression in a dose-dependent manner, which was significantly different from the model group (P<0.01).

[0095] Figure 5 B The results showed that Ang II significantly reduced SIRT3 activity in EPCs (P < 0.001), while SIRT3 activity increased with the increase of rhynchophylline intervention concentration, and the differences were statistically significant compared with the model group (P < 0.01). The results showed that rhynchophylline is a SIRT3 activator and can enhance SIRT3 expression in EPCs.

[0096] (2) Knockout of SIRT3 can reverse the effect of rhynchophylline on improving mitochondrial dysfunction in EPCs

[0097] Figure 6 A results showed that after Ang II intervention, the basal respiratory rate, maximum respiratory rate and ATP generation rate of EPCs were significantly reduced (P < 0.05), and rhynchophylline enhanced the mitochondrial respiration of EPCs and significantly increased the cellular oxygen consumption rate and ATP generation rate (P < 0.05). After knocking out SIRT3 in EPCs, the effect of rhynchophylline on improving the mitochondrial respiration of EPCs was significantly weakened.

[0098] Figure 6 B The results showed that after Ang II intervention, the mitochondrial membrane potential of EPCs was significantly decreased (P < 0.001); Rhynchophylline significantly increased the mitochondrial membrane potential of EPCs, but after knocking out SIRT3 in EPCs, the effect of Rhynchophylline in maintaining the mitochondrial membrane potential of EPCs was significantly weakened.

[0099] (3) Knockout of SIRT3 can reverse the effect of rhynchophylline on EPCs damage

[0100] Figure 7 A The results showed that after knocking out SIRT3 in EPCs, the effect of rhynchophylline on improving the proliferation ability of EPCs was significantly weakened.

[0101] Figure 7 The results in B showed that after knocking out SIRT3 in EPCs, the effect of rhynchophylline on improving the migration ability of EPCs was significantly weakened.

[0102] Figure 7 C The results showed that after knocking out SIRT3 in EPCs, the effect of rhynchophylline on improving the tube-forming ability of EPCs was significantly weakened.

[0103] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of rhynchophylline in the preparation of histone deacetylase 3 (SIRT3) activators.

2. The use according to claim 1, characterized in that: Rhynchophylline, as a SIRT3 activator, enhances SIRT3 expression in EPCs.

3. Application of SIRT3 activators in the preparation of drugs for the treatment of vascular endothelial damage caused by hypertension.

4. Use of rhynchophylline in the preparation of drugs for preventing and / or treating vascular endothelial damage caused by hypertension.

5. The use according to claim 4, characterized in that: Rhynchophylline can maintain the morphology and structural integrity of the vascular endothelium, regulate vasoconstriction and vasodilation factors, and improve endothelium-dependent vasodilation function; it can inhibit vascular fibrosis by inhibiting collagen deposition, delay the process of vascular fibrosis, and thus reduce vascular wall damage.

6. The use according to claim 4, characterized in that: Rhynchophylline can enhance EPCs mitochondrial respiration, increase cellular oxygen consumption rate and ATP production rate.

7. The use according to claim 4, characterized in that: Rhynchophylline can increase the mitochondrial membrane potential of EPCs.

8. The use according to claim 4, characterized in that: The dosage for the efficacy experiment of treating essential hypertension rat model was 0.862 mg kg -1 ·d -1 .

9. The use according to claim 4, characterized in that: The dosage for treating EPCs dysfunction model in vitro was 100 μmol·L -1 .

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