Application of delta-Viniferin in preparation of medicine for preventing or treating metabolic cardiovascular diseases

Abstract

The invention relates to an application of delta-Viniferin in preparation of a medicine for preventing or treating metabolic cardiovascular diseases. Delta-Viniferin reduces endothelial dysfunction by inhibiting vascular endothelial cell senescence and inflammation related gene / protein expression level, and defines the antihypertensive effect of delta-Viniferin; delta-Viniferin improves the contraction function of vascular smooth muscle by inhibiting the expression level of gene / protein related to phenotype transformation of vascular smooth muscle cells, meanwhile, the in-vivo level remarkably relieves degradation of elastic fibers of vascular walls and rupture of intima of aortic walls, and the effect of the delta-Viniferin on preventing and treating aortic dissection is defined; delta-Viniferin defines the anti-myocardial fibrosis effect of delta-Viniferin by inhibiting the myocardial fibroblast fibrosis related gene / protein expression level; the delta-Viniferin promotes the browning of white fat by targeting an SIRT1-FOXO1 axis and increases the energy consumption, so that the effect of the delta-Viniferin on resisting obesity is defined. In conclusion, it is proved that the compound delta-Viniferin has the effect of protecting various metabolic cardiovascular diseases and can be used for preparing the medicine for preventing and treating the metabolic cardiovascular diseases.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of delta-Viniferin in preparing medicines for preventing or treating metabolic cardiovascular diseases. Background Art

[0002] Metabolic cardiovascular disease (CMD) is a general term for cardiovascular diseases or metabolic diseases caused by metabolic risk factors (dysglucose metabolism, dyslipidemia, hypertension, obesity, etc.), and has become a major public health problem that seriously threatens people's health. Unhealthy eating habits and lifestyles and an aging population have further accelerated the progression of metabolic cardiovascular disease. Metabolic cardiovascular disease involves a wide range of disciplines. In addition to being closely related to cardiovascular and endocrine and metabolic diseases, it also involves nephrology, neurology, geriatrics, etc. For this complex interdisciplinary chronic disease, new requirements and new methods must be put forward for diagnosis, treatment and management models. However, there is currently no treatment recommended by clinical guidelines or expert consensus, and its pathophysiology and molecular mechanisms are unclear, which greatly limits the development of effective therapeutic drugs. Therefore, in-depth analysis of the pathophysiology and molecular biological mechanisms of metabolic cardiovascular disease, discovery of new biomarkers / new drug targets, and the development of early diagnostic reagents / effective therapeutic drugs on this basis have important scientific significance and clinical application value for solving this major global public health problem.

[0003] Resveratrol dimer is also called viniferin. There are many structural types, such as ε-Viniferin and δ-Viniferin (resveratrol dehydrogenated dimer), etc. It is mainly found in grapes. Resveratrol dimer is formed by the oxidation of resveratrol by 4-hydroxystilbene peroxidase. All these types of viniferin show important biological activities and have many possible applications in clinical research and future drug development. They may become the most effective nutritional supplements and auxiliary drugs. It has been reported that ε-Viniferin has strong antioxidant, anti-inflammatory, anti-diabetic and anti-neurodegenerative disease activities, among which the biological activities in antioxidant and antibacterial aspects are stronger than resveratrol.

[0004] Patent CN115108989B discloses a resveratrol derivative and its preparation method and application, wherein the resveratrol derivatives are SS-A6, SS-A13 and SS-A14. It also provides the application of resveratrol derivatives in the preparation of drugs for preventing or treating bacterial infections and as inhibitors of Salmonella T3SS virulence proteins. The resveratrol derivatives SS-A6, SS-A13 and SS-A14 provided in the application can effectively inhibit the secretion of Salmonella T3SS virulence proteins, and can significantly reduce the invasion of Salmonella on host cells, which is of great significance for the discovery of new anti-bacterial virulence drugs with broad spectrum, high efficiency and low toxicity, and can be used to prepare drugs for preventing or treating bacterial infections. In addition, the resveratrol derivatives SS-A6, SS-A13 and SS-A14 have concentration gradient dependence on the secretion of Salmonella T3SS virulence proteins SipA / B / C / D, and SS-A6 is the most active T3SS inhibitor.

[0005] Patent CN115246802B discloses a class of viniferin derivatives, their preparation methods, pharmaceutical compositions and uses, and specifically relates to a class of 2,3-diaryl-5-phenylvinylbenzofuran-type viniferin derivatives and their medically acceptable salts, as well as pharmaceutical compositions of such compounds and their use in the preparation of drugs for preventing, treating or assisting in the treatment of inflammatory immune-related diseases.

[0006] Patent CN103275044B discloses the use of resveratrol dimer (7R, 8R)-trans-δ-viniferin for lowering blood sugar.

[0007] However, there are few studies on the effects of vinblastine and its derivatives and experimental drugs disclosed in the prior art on metabolic cardiovascular diseases, etc. It is still necessary to continue to research and develop new vinblastine and its derivatives to meet people's growing medical and health needs. Summary of the invention

[0008] Based on the current situation that there is a lack of effective drugs for preventing and treating metabolic cardiovascular diseases in the prior art, the present invention provides a use of δ-Viniferin (resveratrol dehydrogenated dimer) in the preparation of drugs for preventing and treating metabolic cardiovascular diseases.

[0009] The purpose of the present invention can be achieved by the following technical solutions:

[0010] The present invention first provides the use of resveratrol dehydrogenated dimer δ-Viniferin or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing or treating metabolic cardiovascular diseases. The chemical formula of the δ-Viniferin is shown in Formula I:

[0011]

[0012] In one embodiment of the present invention, the δ-Viniferin or a pharmaceutically acceptable salt thereof is a racemate (±)-δ-Viniferin or (R,R)-δ-Viniferin or (S,S)-δ-Viniferin, and its structure is shown in Formula II or Formula III or Formula IV:

[0013]

[0014] In one embodiment of the present invention, the metabolic cardiovascular diseases include hypertension, aortic dissection, myocardial fibrosis, and obesity.

[0015] In one embodiment of the present invention, the δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for reducing endothelial dysfunction to resist hypertension by inhibiting the expression level of genes / proteins related to vascular endothelial cell senescence and inflammation.

[0016] In one embodiment of the present invention, the δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for preventing and treating aortic dissection by inhibiting the expression level of genes / proteins related to phenotype conversion of vascular smooth muscle cells, improving the contractile function of vascular smooth muscle, and alleviating the degradation of vascular wall elastic fibers and rupture of the aortic intima.

[0017] In one embodiment of the present invention, the δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for preventing and treating myocardial fibrosis by inhibiting the expression level of fibrosis-related genes / proteins.

[0018] In one embodiment of the present invention, the δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for promoting browning of white fat and increasing energy consumption to resist obesity by targeting the SIRT1-FOXO1 axis.

[0019] In one embodiment of the present invention, the drug further comprises a pharmaceutically acceptable excipient and / or carrier.

[0020] In one embodiment of the present invention, the dosage form of the drug includes tablets, capsules, soft capsules, granules, powders, pills, gel candies, powders, oral liquids or drops.

[0021] In one embodiment of the present invention, the administration route of the drug includes an enteral administration route or a parenteral administration route.

[0022] The present invention studies have demonstrated that δ-Viniferin can play a preventive and therapeutic role in different metabolic cardiovascular diseases (hypertension, aortic dissection, myocardial fibrosis, obesity) through different drug action mechanisms.

[0023] Specifically, the present invention establishes an Ang II-induced rat aortic endothelial cell (RAECs) aging and inflammation model, and detects the effects of δ-Viniferin on vascular endothelial aging and inflammatory phenotypes by Western blot and RT-qPCR, thereby clarifying the potential of δ-Viniferin in preventing and treating hypertension. RT-qPCR results show that δ-Viniferin significantly alleviates the levels of aging genes (Lmnb1) and inflammatory genes (Nos3, Il1β) induced by Ang II; western blot results show that δ-Viniferin also significantly alleviates the expression of aging marker proteins (γH2AX), endothelial tight junction proteins (ZO-1) and inflammatory marker proteins (NLRP3, IL-1β) induced by Ang II.

[0024] The present invention establishes IL-1β-induced rat aortic smooth muscle cell (VSMCs) phenotypic transformation and BAPN-induced aortic dissection mouse model, and detects the effect of δ-Viniferin on VSMCs phenotypic transformation and aortic dissection formation by Western blot, RT-qPCR, H&E staining and other methods, thereby clarifying the potential of δ-Viniferin in preventing and treating aortic dissection. RT-qPCR results show that IL-1β induces a decrease in the expression of VSMCs contractile phenotype genes (Acta2, Myh11, Cnn1), while an increase in the expression of inflammatory genes (Nlrp3), and δ-Viniferin treatment can alleviate VSMCs phenotypic transformation and inflammatory phenotype. Western blot results show that δ-Viniferin significantly increases the expression of IL-1β-induced VSMCs contractile phenotype marker proteins (MYH11, CNN1, α-SMA) and Sirt1 protein. In addition, H&E staining results showed that δ-Viniferin could alleviate BAPN-induced vascular wall elastic fiber degradation and aortic wall intima rupture, and further reduce the influence of aortic metalloproteinase gene levels and alleviate BAPN-induced contractile phenotype marker protein (MYH11, α-SMA) expression.

[0025] The present invention establishes a TGFβ1 / Ang II-induced myocardial fibrosis model, detects the effect of δ-Viniferin on the fibrosis phenotype by Western blot, RT-qPCR and immunofluorescence staining, and then clarifies the potential of δ-Viniferin in resisting myocardial fibrosis. The results show that δ-Viniferin significantly reduces the levels of TGFβ1-induced fibrosis genes (Fn, Mmp2, Mmp9) and TGFβ1-induced fibrosis marker proteins (FN, α-SMA, Collagen I, MMP2, MMP9) expression. In addition, the results of immunofluorescence staining also show that δ-Viniferin can reduce the expression of Ang II-induced fibrosis phenotype protein (α-SMA).

[0026] The present invention establishes an in vivo and in vitro white fat browning model, and detects the effect of δ-Viniferin on the white fat browning phenotype by Western blot, RT-qPCR and other methods, thereby clarifying the potential of δ-Viniferin to promote white fat browning and resist obesity. The results of in vivo and in vitro experiments show that δ-Viniferin can significantly increase the expression of white fat browning marker (UCP1) and δ-Viniferin potential target SIRT1 and transcription factor FOXO1 mRNA / protein.

[0027] In conclusion, δ-Viniferin can be used to prepare drugs for preventing and treating metabolic cardiovascular diseases.

[0028] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0029] The present invention explores the protective effect of the compound δ-Viniferin on metabolic cardiovascular diseases by establishing various in vivo and in vitro metabolic cardiovascular disease models such as hypertension, aortic dissection, myocardial fibrosis, obesity, etc.

[0030] The results showed that δ-Viniferin reduced endothelial dysfunction by inhibiting the expression levels of genes / proteins related to senescence and inflammation of vascular endothelial cells, thus clarifying its anti-hypertensive effect; δ-Viniferin improved the contractile function of vascular smooth muscle by inhibiting the expression levels of genes / proteins related to phenotype conversion of vascular smooth muscle cells, and significantly alleviated the degradation of vascular wall elastic fibers and rupture of the aortic intima at the in vivo level, thus clarifying its role in preventing and treating aortic dissection; δ-Viniferin inhibited the expression levels of genes / proteins related to fibrosis of cardiac fibroblasts, thus clarifying its anti-myocardial fibrosis effect; δ-Viniferin promoted the browning of white fat and increased energy consumption by targeting the SIRT1-FOXO1 axis, thus clarifying its anti-obesity effect.

[0031] In summary, it is proved that the compound δ-Viniferin has the effect of protecting against a variety of metabolic cardiovascular diseases and can be used to prepare drugs for preventing and treating metabolic cardiovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 : Effects of δ-Viniferin on Ang II-induced senescence and inflammation in RAECs.

[0033] Among them, (A) CCK8 kit was used to detect the effect of δ-Viniferin on RAECs vitality; (B) RT-qPCR method was used to detect the effect of δ-Viniferin on the levels of senescence genes (Lmnb1) and inflammatory genes (Nos3, Il1β) in RAECs induced by Ang II; (C) Western blot method was used to detect the effect of δ-Viniferin on the expression of senescence marker proteins (γH2AX), endothelial tight junction proteins (ZO-1) and inflammatory marker proteins (NLRP3, IL-1β) in RAECs induced by Ang II. Ctrl, normal control group; δ-Vini, δ-Vini (5μM) single administration group; Ang II+Veh, model group; Ang II+δ-Vini, model+δ-Vini (5μM) group. *p<0.05, **p<0.01, ***p<0.001.

[0034] Figure 2 :Effect of δ-Viniferin on phenotypic transformation of VSMCs induced by IL-1β.

[0035] Among them, (A) CCK8 kit was used to detect the effect of δ-Viniferin on VSMCs vitality; (B) RT-qPCR method was used to detect the effect of δ-Viniferin on the levels of IL-1β-induced VSMCs contraction phenotype genes (Acta2, Myh11, Cnn1) and inflammatory genes (Nlrp3); (C) Western blot method was used to detect the effect of δ-Viniferin on IL-1β-induced VSMCs contraction phenotype marker proteins (MYH11, CNN1, α-SMA) and Sirt1 protein expression. Ctrl, normal control group; δ-Vini, δ-Vini (2μM) single administration group; IL-1β+Veh, model group; IL-1β+δ-Vini, model+δ-Vini (2μM) group. *p<0.05, **p<0.01, ***p<0.001.

[0036] Figure 3 : Effects of δ-Viniferin on BAPN-induced aortic dissection mouse model.

[0037] Among them, (A) H&E staining was used to observe the changes in the cross-section of blood vessels in each group. (B) The RT-qPCR method was used to detect the effect of δ-Viniferin on the levels of metalloproteinase genes (Mmp2, Mmp9) induced by BAPN; (C) The western blot method was used to detect the effect of δ-Viniferin on the expression of contractile phenotype marker proteins (MYH11, α-SMA) induced by BAPN. Sham: sham operation group; BAPN+Vehicle: aortic dissection model group; BAPN+δ-Vini (10mg / kg / day), aortic dissection model + δ-Vini (10mg / kg / day) administration group; BAPN+δ-Vini (20mg / kg / day), aortic dissection model + δ-Vini (20mg / kg / day) administration group. *p<0.05, **p<0.01, ***p<0.001.

[0038] Figure 4 :Effects of δ-Viniferin on TGFβ1-induced myocardial fibrosis.

[0039] Among them, (A) CCK8 kit was used to detect the effect of δ-Viniferin on the viability of rat neonatal primary cardiac fibroblasts (NRCFs); (B) RT-qPCR method was used to detect the effect of δ-Viniferin on the levels of TGFβ1-induced fibrosis genes (Fn, Mmp2, Mmp9); (C) Western blot method was used to detect the effect of δ-Viniferin on the expression of TGFβ1-induced fibrosis marker proteins (FN, α-SMA, Collagen I, MMP2, MMP9); (D) Ang II was used to induce another fibrosis cell model, and immunofluorescence staining was used to detect the effect of Ang II-induced fibrosis phenotype protein (α-SMA). Ctrl, normal control group; TGFβ1+Veh, model group; TGFβ1+δ-Vini, model+δ-Vini (5μM) group; Ang II+Veh, model group; Ang II+δ-Vini, model+δ-Vini (5μM) group. *p<0.05, **p<0.01, ***p<0.001.

[0040] Figure 5 : Effect of δ-viniferin on the browning of white adipose tissue.

[0041] Among them, (A) the CCK8 kit was used to detect the effect of δ-Viniferin on the vitality of preadipocytes 3T3-L1; (B) the western blot method was used to detect the effect of δ-Viniferin on the expression of white fat browning marker protein (UCP1) and the expression of δ-Viniferin potential targets SIRT1 and transcription factor FOXO1 protein; (C) Schematic diagram of in situ injection of δ-Viniferin into inguinal adipose tissue (iWAT) in vivo; (D) mRNA was extracted from iWAT, and the RT-qPCR method was used to detect the effect of δ-Viniferin in situ injection into iWAT on the levels of white fat browning marker gene (Ucp1) and potential target Sirt1 gene; (E) Total protein was extracted from iWAT, and the western blot method was used to detect the effect of δ-Viniferin on the expression of white fat browning marker protein (UCP1) and the expression of δ-Viniferin potential targets SIRT1 and transcription factor FOXO1 protein. Control, normal control group; δ-Vini, δ-Vini (5μM) alone administration group; Browning, model group; Browning+δ-Vini, model+δ-Vini (5μM) group. *p<0.05, **p<0.01, ***p<0.001. DETAILED DESCRIPTION

[0042] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

[0043] Example 1: Effects of δ-Viniferin on Ang II-induced RAECs senescence and inflammation.

[0044] In this example, an Ang II-induced rat aortic endothelial cell (RAECs) senescence and inflammation model was established, and the effects of δ-Viniferin on vascular endothelial senescence and inflammatory phenotypes were detected by Western blot and RT-qPCR to further illustrate the potential of δ-Viniferin in preventing and treating hypertension.

[0045] Rat primary aortic endothelial cells (RAECs) were cultured in DMEM medium containing 10% fetal bovine serum at 37°C and 5% CO. 2The cells were cultured in an incubator and passaged when the confluence reached 80-90%. The Ctrl group (no drug intervention, no Ang II), Ang II+Veh group (model group, no drug intervention, 1μM Ang II stimulation for 48h, drug solvent added), and Ang II+δ-Vini group (δ-Vini, 5μM, pre-added 1h in advance and then stimulated with Ang II) were set up. RAECs senescence and inflammation models were induced by Ang II (1μM, 48h) culture, and total cell protein and total mRNA were extracted. Western Blot was used to determine the expression of senescence marker proteins (γH2AX), endothelial tight junction proteins (ZO-1) and inflammatory marker proteins (NLRP3, IL-1β), and RT-qPCR experiments were used to determine the changes in the levels of aging genes (Lmnb1) and inflammatory genes (Nos3, Il1β). Results are shown in Figure 1 As shown, RT-qPCR results showed that δ-Viniferin significantly alleviated the levels of aging genes (Lmnb1) and inflammatory genes (Nos3, Il1β) induced by Ang II; western blot results showed that δ-Viniferin also significantly alleviated the expression of aging marker proteins (γH2AX), endothelial tight junction proteins (ZO-1) and inflammatory marker proteins (NLRP3, IL-1β) induced by Ang II. The above results prove that the δ-Viniferin has obvious anti-aging and anti-inflammatory effects and can be used to prepare drugs for the prevention and treatment of hypertension.

[0046] Example 2: Effects of δ-Viniferin on IL-1β-induced phenotypic transformation of VSMCs and BAPN-induced aortic dissection.

[0047] In this example, IL-1β-induced rat aortic smooth muscle cell (VSMCs) phenotypic transformation and BAPN-induced aortic dissection mouse model were established, and the effects of δ-Viniferin on VSMCs phenotypic transformation and aortic dissection formation were detected by Western blot, RT-qPCR, H&E staining and other methods, thereby illustrating the potential of δ-Viniferin in preventing and treating aortic dissection.

[0048] Primary rat aortic smooth muscle cells were cultured in DMEM medium containing 10% fetal bovine serum at 37°C and 5% CO. 2Cultured in an incubator, the cells were passaged when the confluence reached 80-90%. Ctrl, normal control group; δ-Vini, δ-Vini (2μM) single administration group; IL-1β+Veh, model group; IL-1β+δ-Vini, model+δ-Vini (2μM) group. Total cell protein and total mRNA were extracted from the above groups, and the RT-qPCR method was used to detect the effect of δ-Viniferin on the levels of IL-1β-induced VSMCs contraction phenotype genes (Acta2, Myh11, Cnn1) and inflammatory genes (Nlrp3); the western blot method was used to detect the effect of δ-Viniferin on the expression of IL-1β-induced VSMCs contraction phenotype marker proteins (MYH11, CNN1, α-SMA) and Sirt1 protein. Results are shown in Figure 2 As shown in the figure, RT-qPCR results showed that IL-1β induced a decrease in the expression of VSMCs contractile phenotype genes (Acta2, Myh11, Cnn1), while an increase in the expression of inflammatory genes (Nlrp3). δ-Viniferin treatment can alleviate the phenotypic transformation and inflammatory phenotype of VSMCs. Western blot results showed that δ-Viniferin significantly increased the expression of IL-1β-induced VSMCs contractile phenotype marker proteins (MYH11, CNN1, α-SMA) and Sirt1 protein.

[0049] 3-week-old wild-type C57BL / 6J mice were used to establish an aortic dissection mouse model using β-aminopropionitrile monofumarate (BAPN) (A3134, Sigma-Aldrich). β-aminopropionitrile (BAPN) is a lysyl oxidase inhibitor that destroys the integrity of the arterial wall structure by inhibiting the cross-linking of collagen and elastin. Researchers have reported that continuous administration of BAPN to C57BL / 6J mice in drinking water can cause degeneration of the arterial media, leading to AD and aneurysm formation. BAPN (1.0 g / kg / day) was dissolved in drinking water and continuously consumed for 4 weeks to induce the formation of an aortic dissection model. δ-Viniferin was dissolved in water, mixed, and administered to mice daily by gavage. The mice were randomly divided into Sham: sham operation group; BAPN+Vehicle: aortic dissection model group; BAPN+δ-Vini (10mg / kg / day), aortic dissection model + δ-Vini (10mg / kg / day) administration group; BAPN+δ-Vini (20mg / kg / day), aortic dissection model + δ-Vini (20mg / kg / day) administration group. After 28 days, the aorta was collected, one part was fixed with 4% paraformaldehyde, embedded in paraffin sections, and H&E staining was used to observe the vascular morphology; the other part was stored in liquid nitrogen for subsequent Western Blot and RT-qPCR detection; the results are shown in Figure 3 H&E staining results showed that δ-Viniferin could alleviate BAPN-induced degradation of vascular wall elastic fibers and intimal rupture of aortic wall, and further reduce the influence of aortic metalloproteinase gene levels and alleviate the expression of contractile phenotype marker proteins (MYH11, α-SMA) induced by BAPN.

[0050] The above results prove that the δ-Viniferin has a significant effect of alleviating the phenotypic transformation of VSMCs and the formation of aortic dissection induced by BAPN, and can be used to prepare drugs for preventing and treating aortic dissection.

[0051] Example 3: Effects of δ-Viniferin on TGFβ1-induced myocardial fibrosis.

[0052] In this example, a TGFβ1-induced myocardial fibrosis model was established, and the effect of δ-Viniferin on the fibrotic phenotype was detected by Western blot, RT-qPCR, and immunofluorescence staining, thereby illustrating the potential of δ-Viniferin in resisting myocardial fibrosis.

[0053] Isolation and extraction of neonatal rat cardiac fibroblasts (NRCFs): prepare 6-8 SD neonatal rats that are 24 hours old, soak them in 75% alcohol for a few seconds for disinfection, fix the neonatal rat with one hand, and use sterile surgical scissors with the other hand to cut the chest cavity, gently squeeze the chest cavity of the suckling rat, and then quickly remove the heart and put it into the pre-cooled PBS buffer containing double antibodies. Use forceps to gently squeeze the heart to remove the blood, and wash it 3 times. Remove the surrounding connective tissue, right ventricle and atrium, and only take the left ventricle. Then cut the left ventricle into about 1mm with surgical scissors. 3 Collect all tissue blocks into a conical flask. Add 5 ml of 0.25% trypsin for digestion, place the conical flask in a 37°C water bath to heat and stir evenly. Digest for about 5 minutes, when the supernatant is slightly turbid, use a pipette to collect the digestion solution into a centrifuge tube and add an equal amount of complete culture medium to terminate the digestion. Add 5 ml of 0.25% trypsin to the conical flask again to continue digestion. Repeat this process about 8 times to completely digest the tissue blocks. Centrifuge the collected digestion solution at 1000 rpm for 3 minutes, discard the supernatant, resuspend it with complete culture medium and transfer it to a 10 mm large dish. The extracted cell suspension is placed at 37°C, CO 2 After 1.5 hours of culture in a constant temperature incubator, a large number of fibroblasts adhered to the wall. The supernatant culture medium was discarded, and the cells were washed with PBS, and 10 ml of new complete culture medium was added to continue the culture. When the cells filled the large dish, they could be passaged and identified. Set Ctrl, normal control group; TGFβ1+Veh, model group; TGFβ1+δ-Vini, model + δ-Vini (5μM) group. Total cell protein and total mRNA were extracted from the above groups, and the RT-qPCR method was used to detect the effect of δ-Viniferin on the levels of TGFβ1-induced fibrosis genes (Fn, Mmp2, Mmp9); the western blot method was used to detect the effect of δ-Viniferin on the expression of TGFβ1-induced fibrosis marker proteins (FN, α-SMA, Collagen I, MMP2, MMP9); Ang II was used to induce another fibrosis cell model, and the groups were set as Ctrl, normal control group; Ang II+Veh, model group; Ang II+δ-Vini, model+δ-Vini (5μM) group, and immunofluorescence staining was used to detect the effect of Ang II-induced fibrosis phenotype protein (α-SMA). The results are shown in Figure 4The results showed that δ-Viniferin significantly reduced the levels of TGFβ1-induced fibrosis genes (Fn, Mmp2, Mmp9) and the expression of TGFβ1-induced fibrosis marker proteins (FN, α-SMA, Collagen I, MMP2, MMP9). In addition, immunofluorescence staining results also showed that δ-Viniferin could reduce the expression of Ang II-induced fibrosis phenotype protein (α-SMA).

[0054] The above results prove that the δ-Viniferin has a significant effect on resisting myocardial fibrosis phenotype and can be used to prepare drugs for preventing and treating myocardial fibrosis.

[0055] Example 4: Effects of δ-Viniferin on white fat browning and obesity.

[0056] In this example, an in vivo and in vitro white fat browning model was established, and the effect of δ-Viniferin on the white fat browning phenotype was detected by Western blot, RT-qPCR and other methods, thereby illustrating the potential of δ-Viniferin to promote white fat browning and thus resist obesity.

[0057] Inducing white fat browning model in vitro. First, 3T3-L1 preadipocytes were inoculated into the well plate. On the second day after contact inhibition (designated as day 0), the differentiation medium (DMEM+10% FBS+1μg / ml insulin+1μM dexamethasone+0.5mM 3-isobutyl-1-methylxanthine) was changed for 48 hours, and then the maintenance medium (DMEM+10% FBS+1μg / ml insulin) was changed for 48 hours. Then, the culture medium (DMEM+10% FBS+corresponding drugs) was changed for 48 hours, and CL316243 was added for 48 hours to obtain the white fat browning model. δ-Viniferin intervention was given starting on day 0. Control, normal control group; δ-Vini, δ-Vini (5μM) single administration group; Browning, model group; Browning+δ-Vini, model+δ-Vini (5μM) group. Total cell protein was extracted from the above groups, and the effect of δ-Viniferin on the expression of white fat browning marker protein (UCP1) and the expression of δ-Viniferin potential targets SIRT1 and transcription factor FOXO1 was detected by western blot. In vivo, δ-Viniferin was injected into inguinal adipose tissue (iWAT) in situ, and iWAT was extracted to extract mRNA. RT-qPCR was used to detect the effect of δ-Viniferin in situ injection into iWAT on the level of white fat browning marker gene (Ucp1) and potential target Sirt1 gene. Total protein was extracted, and the effect of δ-Viniferin on the expression of white fat browning marker protein (UCP1) and the expression of δ-Viniferin potential targets SIRT1 and transcription factor FOXO1 protein was detected by western blot. The results are shown in Figure 2. Figure 5 The results showed that δ-Viniferin can significantly increase the expression of white fat browning marker (UCP1) and δ-Viniferin potential target SIRT1 and transcription factor FOXO1 mRNA / protein at both in vivo and in vitro levels. The above results prove that the δ-Viniferin has the effect of promoting the browning of white fat, and thus has the potential to become a drug for the preparation of prevention and treatment of obesity.

[0058] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. Use of δ-Viniferin or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing or treating metabolic cardiovascular diseases, wherein the chemical formula of the δ-Viniferin is as shown in Formula I:

2. The use according to claim 1, characterized in that: The δ-Viniferin or its pharmaceutically acceptable salt is a racemate (±)-δ-Viniferin or (R,R)-δ-Viniferin or (S,S)-δ-Viniferin, and its structure is shown in Formula II or Formula III or Formula IV:

3. The use according to claim 1, characterized in that: The metabolic cardiovascular diseases include hypertension, aortic dissection, myocardial fibrosis, and obesity.

4. The use according to claim 3, characterized in that: The δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for reducing endothelial dysfunction to resist hypertension by inhibiting the expression level of vascular endothelial cell senescence and inflammation-related genes / proteins.

5. The use according to claim 3, characterized in that: The δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for preventing and treating aortic dissection by inhibiting the expression level of genes / proteins related to phenotype conversion of vascular smooth muscle cells, improving the contractile function of vascular smooth muscle, and relieving the degradation of vascular wall elastic fibers and rupture of the aortic wall intima.

6. The use according to claim 3, characterized in that: The use of the delta-Viniferin or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating myocardial fibrosis by inhibiting the expression level of fibrosis-related genes / proteins.

7. The use according to claim 3, characterized in that: The δ-Viniferin or a pharmaceutically acceptable salt thereof is used in the preparation of a drug for promoting the browning of white fat and increasing energy consumption to resist obesity by targeting the SIRT1-FOXO1 axis.

8. The use according to claim 1, characterized in that: The drug also includes pharmaceutically acceptable excipients and / or carriers.

9. The use according to claim 1, characterized in that: The dosage form of the drug includes tablets, capsules, soft capsules, granules, powders, pills, gel candies, powders, oral liquids or drops.

10. The use according to claim 1, characterized in that: The administration route of the drug includes a gastrointestinal administration route or a parenteral administration route.

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