Application of lycium ruthenicum anthocyanin in preparation of antihypertensive products

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of lycium ruthenicum anthocyanin in preparation of a blood pressure lowering product. The invention provides application of lycium ruthenicum anthocyanin in preparation of a product for reducing blood pressure. The lycium ruthenicum anthocyanin is found to have an antihypertensive effect, can protect target organs such as heart and kidney, alleviate organ injury and reduce TG and LDL levels so as to reduce the risk of cardiovascular diseases, and is in direct proportion to dosage. The lycium ruthenicum anthocyanin has a pharmacodynamic mechanism of lowering blood pressure, lowering the level of angiotensin converting enzyme (ACE), reducing secretion of angiotensin II (AngII) and aldosterone (ALD), and regulating excessive activation of an RAAS system; the level of nitric oxide (NO) is improved, and vascular tension is reduced; the method is related to reduction of the thickness of the middle layer of the vascular intima and improvement of aortic remodeling, and has the advantage of multi-link and multi-target integral adjustment.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and particularly relates to the application of anthocyanins from lycium barbarum in the preparation of blood pressure lowering products. Background Art

[0002] Hypertension refers to the pressure on the blood vessel wall caused by blood flowing in the blood vessels that is higher than normal. It is one of the most common cardiovascular diseases and an important risk factor for diseases such as stroke, coronary heart disease, and heart failure. In the absence of antihypertensive drugs, hypertension can be diagnosed when three blood pressure values ​​measured on different days are higher than normal, that is, systolic blood pressure ≥140mmHg, and (or) diastolic blood pressure ≥90mmHg. Hypertension is currently the most common disease with a very high incidence and growth rate, which seriously endangers human health. Nowadays, people are paying more and more attention to the treatment of hypertension.

[0003] However, the existing drugs for treating hypertension are mainly Western medicines. Although they have significant antihypertensive effects, they also have great side effects and limited protective effects on the vascular endothelium of hypertensive patients. Long-term use of these synthetic drugs will produce some adverse reactions, such as dry cough, taste disorders, rashes and allergic reactions. Therefore, it is particularly necessary to find drugs with significant antihypertensive effects, few side effects and protection of the vascular endothelium of hypertensive patients. Summary of the invention

[0004] The purpose of the present invention is to provide an application of anthocyanins from black wolfberry in the preparation of blood pressure lowering products, enrich the types of blood pressure lowering drugs, have significant blood pressure lowering effects, and have few side effects.

[0005] The present invention provides the use of anthocyanins from black wolfberry in the preparation of a blood pressure lowering product;

[0006] The black wolfberry anthocyanidins include petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-O-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside, malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside;

[0007] The petunidin-3-O-rutin (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanins in the black wolfberry.

[0008] Preferably, the blood pressure lowering product includes a product for preventing and / or treating cardiovascular diseases.

[0009] Preferably, the cardiovascular disease products include hypertension and / or hyperlipidemia.

[0010] Preferably, the blood pressure lowering product includes a product that prevents vascular remodeling and / or reduces vascular tension.

[0011] Preferably, the prevention of vascular remodeling comprises reducing the thickness of the vascular intima-media layer;

[0012] The reduction of vascular tone includes increasing vascular nitric oxide.

[0013] Preferably, the blood pressure lowering product includes a product for improving abnormal lipid metabolism.

[0014] Preferably, the lipid metabolism index includes one or more of total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein and arteriosclerosis index.

[0015] Preferably, the blood pressure lowering product includes a product that improves the oxidative stress response of hepatocytes and / or regulates the overactivation of the renin-angiotensin-aldosterone system.

[0016] The present invention also provides a blood pressure lowering product, wherein the drug effect or effective component of the blood pressure lowering product is anthocyanidin from black wolfberry;

[0017] The black wolfberry anthocyanidins include petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-O-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside, malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside;

[0018] The petunidin-3-O-rutin (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanins in the black wolfberry;

[0019] The products include medicines or health foods.

[0020] Preferably, the product is formulated as a single-use dosage form with 115-462 mg of Lycium ruthenicum anthocyanins per dose.

[0021] Beneficial effects:

[0022] The invention provides application of anthocyanidin of black wolfberry in preparing a blood pressure lowering product; the anthocyanidin of black wolfberry comprises petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-0-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside and malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside; the petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanidin of black wolfberry. The present invention found that the anthocyanidin of black wolfberry has a hypotensive effect, can protect target organs such as the heart and kidneys, alleviate their organ damage, reduce TG and LDL levels to reduce the risk of cardiovascular disease, and is proportional to the dose. The pharmacodynamic mechanism of the anthocyanidin of black wolfberry in lowering blood pressure is related to reducing the level of angiotensin converting enzyme (ACE), reducing the secretion of angiotensin II (AngII) and aldosterone (ALD), regulating the overactivation of the renin-angiotensin-aldosterone system; increasing the level of nitric oxide (NO), reducing vascular tension; reducing the thickness of the intima-media layer (IMT) of the blood vessels to improve aortic remodeling, and has the advantages of multi-link and multi-target overall regulation. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0024] Figure 1 The results of systolic blood pressure (SBP) test of mice in different treatment groups; △ indicates P < 0.05 compared with the control group, and * indicates P < 0.05 compared with the model group;

[0025] Figure 2 Diastolic blood pressure (DBP) test results of mice in different treatment groups; △ indicates P < 0.05 compared with the control group, and * indicates P < 0.05 compared with the model group;

[0026] Figure 3 The weight test results of mice in different treatment groups at the last administration; the mean values ​​between different letters are significantly different (P<0.05);

[0027] Figure 4 The results of body weight test of mice in different treatment groups during drug administration;

[0028] Figure 5 The results of serum TG, T-CHO, HDL-CL and DL-C test of mice in different treatment groups; the average values ​​between different letters are significantly different (P<0.05);

[0029] Figure 6 The results of serum lipid index test of mice in different treatment groups; the average values ​​between different letters are significantly different (P<0.05);

[0030] Figure 7 The results of serum ALT, AST and AKP detection in mice of different treatment groups; the average values ​​between different letters are significantly different (P<0.05);

[0031] Figure 8 HE staining results of kidneys of mice in different treatment groups (4X); red arrows / circles indicate inflammatory cell infiltration; blue arrows indicate diffuse fibrous necrosis; yellow arrows indicate tissue cell edema;

[0032] Fig. 9 The results of HE staining of kidneys of mice in different treatment groups (20X); black arrows indicate tissue necrosis; red arrows indicate glomeruli; blue arrows indicate renal interstitial fibroblast proliferation; yellow arrows indicate inflammatory cell infiltration;

[0033] Fig.10 The results of Masson staining of kidneys of mice in different treatment groups (20X); the red arrows indicate blue fiber reticular deposition; the yellow arrows indicate collagen deposition in the glomerular mesangial area;

[0034] Fig.11 The results of serum BUN test of mice in different treatment groups; the mean values ​​between different letters are significantly different P<0.05

[0035] Fig.12 The results of serum CER detection of mice in different treatment groups; the mean values ​​between different letters are significantly different P<0.05

[0036] Fig.13 The results of NO and ET-1 detection in serum of mice in different treatment groups; the mean values ​​between different letters are significantly different (P<0.05);

[0037] Fig.14 HE staining results of aorta vessels of mice in different treatment groups (10X);

[0038] Fig.15 The results of the aortic intima-media thickness test of mice in different treatment groups; the mean values ​​between different letters are significantly different (P<0.05);

[0039] Fig.16 These are the results of the detection of related indicators of the renin-angiotensin-aldosterone system (RAAS system) in mice in different treatment groups; the mean values ​​between different letters are significantly different (P < 0.05);

[0040] Fig.17This is the HPLC analysis chart of anthocyanins from black wolfberry. DETAILED DESCRIPTION

[0041] The present invention provides the use of anthocyanidin of black wolfberry in the preparation of a blood pressure lowering product; the anthocyanidin of black wolfberry includes petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-0-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside, malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside; the petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanidin of black wolfberry. The anthocyanidin of black wolfberry of the present invention is preferably purchased from Qinghai Jinmaiqi Biotechnology Co., Ltd., and the HPLC analysis chart is as follows Fig.17 The product includes medicine and health food. The product of the present invention preferably includes medicine or health food

[0042] In the present invention, the blood pressure lowering product preferably includes a product for preventing and / or treating cardiovascular diseases; the cardiovascular diseases preferably include hypertension and / or hyperlipidemia. The present invention uses Lycium ruthenicum anthocyanidin as an active ingredient, which can improve hypertension and / or hyperlipidemia in a dose-dependent manner.

[0043] In the present invention, the blood pressure lowering product preferably includes a product that prevents vascular remodeling and / or reduces vascular tension. The prevention of vascular remodeling in the present invention preferably includes reducing the thickness of the vascular intima-media layer; the reducing vascular tension preferably includes increasing vascular nitric oxide. The present invention uses Lycium ruthenicum anthocyanidin as an active ingredient, which can reduce the thickness of the vascular intima-media layer induced by L-NAME, prevent aortic remodeling, increase the level of nitric oxide (NO), reduce vascular tension, and has a blood pressure lowering effect.

[0044] In the present invention, the blood pressure lowering product preferably includes a product that improves lipid metabolism abnormalities and lowers blood pressure. The lipid metabolism index of the present invention preferably includes one or more of total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein and arteriosclerosis index, and is further preferably total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein and arteriosclerosis index. The present invention uses black wolfberry anthocyanidin as an active ingredient, which can reduce TG and LDL levels to reduce the risk of cardiovascular disease, which is proportional to the dosage.

[0045] In the present invention, the blood pressure lowering product preferably includes a product that improves the oxidative stress response of hepatocytes and / or regulates the overactivation of the renin-angiotensin-aldosterone system. The present invention uses black wolfberry anthocyanidins as active ingredients, which can protect target organs such as the heart and kidneys of L-NAME-induced hypertensive mice, alleviate their organ damage, reduce the level of angiotensin converting enzyme (ACE), reduce the secretion of angiotensin II (AngII) and aldosterone (ALD), regulate the overactivation of the RAAS system, and has a blood pressure lowering effect.

[0046] The present invention also provides a blood pressure lowering product, the medicinal effect or effective ingredient of the blood pressure lowering product is anthocyanidin of black wolfberry; the anthocyanidin of black wolfberry includes petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-0-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside, malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside; the petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanidin of black wolfberry; the product includes medicine or health food.

[0047] In the present invention, the product is preferably formulated as a single-use dosage form with 115-462 mg of black wolfberry anthocyanidin / dose, and more preferably 231 mg of black wolfberry anthocyanidin / dose. The dosage of black wolfberry anthocyanidin in the product of the present invention is preferably a daily dosage for a person weighing 70 kg.

[0048] In order to further illustrate the present invention, the application of the anthocyanins of Lycium barbarum provided by the present invention in the preparation of blood pressure lowering products is described in detail below in combination with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0049] Data analysis of the embodiments of the present invention was performed by GraphPadprism 6 to perform statistical analysis on antihypertensive activity, biochemical indices and histomorphological changes. One-way ANOVA and Tukey post hoc test were used for statistical analysis. P < 0.05 was statistically significant. Images were digitally recorded and analyzed using NIU image analysis software and ImageJ software to calculate aortic media thickness. All data are expressed as mean ± standard error mean (Mean ± SEM).

[0050] Example 1

[0051] 1. Experimental reagents and instruments

[0052] Reagents:

[0053] NG-nitro-L-arginine methyl ester hydrochloride (L-NAME, ST1555), hematoxylin-eosin stain (C0105M), hydrochloric acid ethanol slow differentiation solution (C0165M), BCA protein concentration determination kit (P0010S) were purchased from Shanghai Biyuntian Biotechnology Co., Ltd.; Masson trichrome stain (ml093341), mouse aldosterone (ALD, ml037530V), angiotensin converting enzyme (ACE, ml002231V), angiotensin II (AngII, ml058193V), endothelial nitric oxide synthase (eNOS, ml063615V), renin (Renin, ml002045V), endothelial ET-1 (ml037530V) ELISA kit was purchased from Shanghai ELISA Biotechnology Co., Ltd.; total cholesterol (T-CHO, A111-1-1), triglyceride (TG, A110-1-4), low-density lipoprotein cholesterol (LDL-C, A113-1-1), high-density lipoprotein cholesterol (HDL-C, A112-1-1), aspartate aminotransferase (AST, C010-2-1), alanine aminotransferase (ALT, C009-2-1), alkaline phosphatase (AKP, A059-2-2), urea nitrogen (BUN, C013-2-1), and creatinine (CRE, C011-2-1) kits were purchased from Nanjing Jiancheng Bioengineering Institute;

[0054] Instruments: intelligent non-invasive sphygmomanometer for mice and rats (BP-2010A, Ruanlong, Beijing), multifunctional live cell imaging microplate reader (Spark cyto 400, Tecan, Austria), UV-visible spectrophotometer (L5S, Instrument and Electrical Analysis, Shanghai), inverted fluorescence microscope (DS-Ri2, Nikon Eclipse, Japan), low-temperature high-speed centrifuge (Centrifuge5424R, Eppendorf, Germany), cryostat (CM 1860UV, Leica, Germany), water purifier (Millipore-QIntegral 3, Merk, Germany), handheld tissue grinder (MY-20, Jingxin, Shanghai).

[0055] 2. Establishment of Mouse Hypertension Model and Grouping and Drug Administration

[0056] (1) Male Kunming (KM) mice, weighing 18±2g, were adaptively fed for one week and randomly divided into a control group and a modeling group. The control group drank distilled water, and the modeling group added 2g / LNG-nitro-L-arginine methyl ester (L-NAME) to drinking water. All mice were allowed to eat freely. Blood pressure was measured at fixed time points using the tail cuff method, and body weight, diet and drinking water were monitored for 6 consecutive weeks. After 6 weeks, the systolic blood pressure (SBP) of the modeling group mice was greater than 140mmHg, indicating that they were hypertensive mice. The modeling was successful with a success rate of 90%.

[0057] (2) The hypertensive mice in the modeling group were randomly divided into a model group and a drug-treated group, and the drugs were administered by gavage according to Table 1 below; the control group was given distilled water, and the mice in each treatment group were given the drug continuously for 8 weeks.

[0058] Table 1 Drug administration method for mice in each treatment group

[0059]

[0060] Test Example 1

[0061] 1. General

[0062] During the administration period of Example 1, no death occurred in the control group, model group, captopril group and drug administration group, and there was no significant difference in diet and water intake.

[0063] 2. Blood pressure test

[0064] Blood pressure was measured using the tail cuff method. All mice were trained in a blood pressure analyzer for one week before modeling and administration. Example 1: Mice were fixed 1 hour after each administration, and systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured after preheating at 37°C for 10 minutes. Each mouse was measured 5 times in a row, once a week. The results are shown in the table below. Figure 1 and Figure 2 shown.

[0065] according to Figure 1 and Figure 2 It can be seen that the systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the mice in the model group were significantly higher than those in the control group. Compared with the model group, the SBP of the mice treated with captopril (positive control) was significantly reduced after 1 week of administration (P < 0.05), and the DBP was significantly reduced after 3 weeks of administration (P < 0.05); the SBP of the high-dose group was significantly reduced after 4 weeks of administration of AEL 30mg / kg and 60mg / kg (P < 0.05) Figure 1 ), although there was a trend in the AEL 15mg / kg group, it was not statistically significant, and had no effect on DBP. AEL can significantly reduce the increase in SBP caused by L-NAME, and has the effect of lowering blood pressure. The antihypertensive effect is positively correlated with the dosage.

[0066] 3. Weight test

[0067] Studies have shown that L-NAME inhibits NOS levels in the hypothalamus and reduces the secretion of ghrelin, thereby reducing body weight and fat content. Long-term induction can cause mice to lose weight significantly. Example 1 Body weight was measured at a fixed time and place after administration, once a week for 8 consecutive weeks. The results are as follows Figure 3 and Figure 4 shown.

[0068] according to Figure 3 and Figure 4 It can be seen that compared with the control group, the final body weight of the model group was significantly reduced, indicating that L-NAME induction for 14 weeks (6 weeks + 8 weeks) can cause significant weight loss in mice; compared with the model group, the captopril group had no effect on the weight loss caused by L-NAME; AEL administration for 8 weeks can significantly improve the weight loss of mice induced by L-NAME, and there are significant differences between the medium and high dose groups in the dose range of 15-60 mg / kg. During the administration period, the body weight of the model group was lower than that of the control group, and the area under the weight change curve (AUC) was significantly reduced; compared with the model group, the AUC of the captopril group was slightly increased, but not statistically significant; the AUC of the 15 mg / kg AEL group was not statistically significant, and the AUC of the 30 mg / kg AEL group and the 60 mg / kg AEL group was significantly increased. AEL can improve the weight loss phenomenon of L-NAME-induced hypertensive mice, which is positively correlated with the dose.

[0069] 4. Visceral system number

[0070] Example 1 Two hours after the last administration, the mice were killed, and the heart, kidney, liver, spleen, and lung were collected and weighed. The organ count of each organ in each treatment group of mice was calculated. The results are shown in Table 2.

[0071] Table 2 Organ counts of mice in different treatment groups

[0072] organ Control L-NAME LAEL MAEL HAEL Captopril heart <![CDATA[0.39±0.03 b ]]> <![CDATA[0.54±0.04 a ]]> <![CDATA[0.44±0.01 b ]]> <![CDATA[0.47±0.03 a ]]> <![CDATA[0.43±0.01 a ]]> <![CDATA[0.43±0.01 b ]]> liver <![CDATA[3.09±0.06 a ]]> <![CDATA[4.07±0.12 a ]]> <![CDATA[3.95±0.17 a ]]> <![CDATA[3.48±0.10 a ]]> <![CDATA[3.27±0.10 a ]]> <![CDATA[3.17±0.11 a ]]> kidney <![CDATA[1.27±0.02 b ]]> <![CDATA[0.83±0.03 a ]]> <![CDATA[1.33±0.07 b ]]> <![CDATA[0.95±0.04 a ]]> <![CDATA[1.28±0.05 b ]]> <![CDATA[1.27±0.04 b ]]> lung <![CDATA[0.91±0.14 a ]]> <![CDATA[1.03±0.24 a ]]> <![CDATA[0.84±0.18 a ]]> <![CDATA[0.85±0.19 a ]]> <![CDATA[0.89±0.16 a ]]> <![CDATA[1.00±0.18 a ]]> spleen <![CDATA[0.26±0.02 a ]]> <![CDATA[0.36±0.06 a ]]> <![CDATA[0.26±0.03 a ]]> <![CDATA[0.25±0.03 a ]]> <![CDATA[0.22±0.01 a ]]> <![CDATA[0.27±0.03 a ]]>

[0073] Note: Different letters in each row indicate that the mean values ​​are significantly different, P < 0.05.

[0074] According to Table 2, the relative weight of the heart of mice induced by L-NAME for 14 weeks (6 weeks + 8 weeks) was significantly higher than that of other groups, which was related to myocardial hypertrophy caused by hypertension; the relative weight of the kidney decreased, which was related to renal damage caused by hypertension, and there was no significant difference in the liver, lung, and spleen. After 8 weeks of administration, the relative weight of the heart and kidney in the captopril group decreased significantly, which could significantly reduce myocardial hypertrophy and renal damage caused by L-NAME; the relative weight of the heart and kidney in the 60mg / kgAEL administration group decreased significantly, while there was no significant difference in the 15mg / kgAEL and 30mg / kgAEL administration groups, which was proportional to the dose. No significant differences were observed in the liver, lung, and spleen of each AEL dose group. L-NAME induced myocardial hypertrophy and renal damage in mice for 14 weeks. AEL administration for 8 weeks can significantly improve myocardial hypertrophy and renal damage in L-NAME-induced hypertensive mice, and the effect is proportional to the dose.

[0075] 5. Serum biochemical index detection

[0076] Example 1 Two hours after the last administration, mice were killed, and blood samples were collected and coagulated at room temperature for 2 hours. The blood samples were centrifuged at 4°C, 3500 rpm / min for 10 minutes, and the serum was separated and stored at 80°C for the detection of serum biochemical indicators.

[0077] (1) Serum total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL-C), and low-density lipoprotein (LDL-C) related to lipid metabolism

[0078] Hypertension and hyperlipidemia are two related cardiovascular risk factors. Therefore, hypertension and dyslipidemia should be controlled to reduce cardiovascular damage. L-NAME-induced hypertensive mice are characterized by high levels of total TC, LDL-C, LDL-C, TG and low levels of HDL-C.

[0079] The present invention detects the changes in serum TC, TG, HDL-C, LDL-C content and related blood lipid indexes related to lipid metabolism, studies the blood lipid levels of mice in each treatment group after 8 weeks of administration, and evaluates the damage of the samples to cardiovascular and cerebrovascular diseases such as atherosclerosis and coronary heart disease. The test results are as follows Figure 5 and Figure 6 As shown in the figure, HDL-C is negatively correlated with the occurrence of coronary heart disease, TG is closely related to the risk of atherosclerotic cardiovascular disease, the larger the AI ​​index value, the more severe the degree of arteriosclerosis, and the higher the risk of cardiovascular and cerebrovascular diseases; serum LDL-C / HDL-C is an important risk factor for coronary heart disease, and TG / HDL-C is used to predict the risk of coronary heart disease; TC / HDL-C as a comprehensive blood lipid index can better express lipid metabolism abnormalities.

[0080] according to Figure 5It can be seen that compared with the control group, the TG, T-CHO, and LDL-C levels of the model group mice increased significantly, and HDL-C decreased significantly, showing obvious dyslipidemia. Compared with the model group, after 8 weeks of administration, the TG and LDL-C levels of the AEL-administered group were significantly reduced, while T-CHO and HDL-C had no effect, showing a dose-dependent manner.

[0081] according to Figure 6 It can be seen that compared with the control group, the arteriosclerosis index (AI), LDL-C / HDL-C, TC / HDL-C, and TG / HDL-C indices of mice in the model group were significantly increased, indicating abnormal lipid metabolism, which leads to a higher risk of arteriosclerosis, coronary heart disease, and death; after 8 weeks of administration, the LDL-C / HDL-C and TG / HDL-C ratios of the AEL-administered group decreased, and the AI ​​index and TC / HDL-C ratio were not affected, showing a dose-dependent manner.

[0082] In summary, captopril can reduce LDL-C and LDL-C / HDL-C, thereby reducing the risk of coronary heart disease in hypertensive mice, but has no improvement on abnormal lipid metabolism; AEL improves vascular sclerosis and reduces the risk of coronary heart disease in hypertensive mice by reducing TC, LDL-C, LDL-C / HDL-C and TG / HDL-C, but has no significant improvement on abnormal lipid metabolism caused by hypertension.

[0083] (2) Serum ALT, AST, and AKP tests related to liver function

[0084] The present invention detected the serum ALT, AST, and AKP levels related to liver function. The results are as follows: Figure 7 As shown. Figure 7 It can be seen that compared with the control group, the AST of the mice in the model group increased significantly, while ALT and AKP did not change significantly. The increase in serum AST content is related to mitochondrial damage of hepatocytes, indicating that continuous administration of L-NAME can induce significant oxidative stress in mouse hepatocytes. Compared with the model group, after 8 weeks of administration, the positive control captopril had no effect on AST; the medium and high dose groups of AEL could significantly reduce AST levels. AEL improved the oxidative stress response of hepatocytes in L-NAME-induced hypertensive mice in a dose-dependent manner.

[0085] 6. Kidney tissue index detection

[0086] Example 1 Mice were killed 2 hours after the last administration, and blood samples were collected and coagulated at room temperature for 2 hours. Serum was separated and used for serum BUN and CER detection. Kidneys were collected and a small portion was immersed in 10% neutral formalin for HE staining and Masson staining. The results are shown in Figure 1. Figures 8 to 12 shown.

[0087] according to Figure 8 and Fig. 9 HE staining results show that after 14 weeks of L-NAME induction (6 weeks of modeling + 8 weeks of drug administration), the morphological structure of renal tissue in the control group was intact, the renal tissue cells were orderly distributed, the morphological structure of glomeruli and renal tubules was intact, and no obvious pathological damage was observed; in the model group, diffuse fibrous necrosis of renal tissue, thickening and enlargement of the basement membrane of the glomerular capsule wall, a large number of inflammatory cells infiltrated, and severe renal damage was observed; captopril reduced necrotic tissue and inflammatory cell infiltration, mild glomerular lesions, and significantly alleviated pathological symptoms, but edema of some tissue cells and hyperplasia of renal interstitial fibroblasts occurred; in the AEL administration group, tissue necrosis and inflammatory cell infiltration were reduced, glomerular volume was normal, and renal pathological damage was significantly improved, which was positively correlated with the dose. The high-dose group was better than the positive control.

[0088] according to Fig.10 The results of Masson staining showed that the control group had normal glomerular and tubular tissue structure, complete morphology and structure, basically no fiber deposition, and no obvious pathological damage. In the model group, more blue fiber deposition appeared in the renal interstitium and renal blood vessels, which were connected into a network (red arrow), and collagen deposition in the glomerular mesangial region (yellow arrow), and obvious fibrosis occurred. In the captopril group, the amount of collagen deposition in the renal interstitium, renal blood vessels, and glomeruli was significantly reduced, and the inflammatory cell infiltration in the kidneys of hypertensive mice was improved. In the AEL group, the collagen deposition in the renal interstitium and renal blood vessels was reduced, and the collagen deposition in the glomerular mesangial region was significantly reduced, which had a certain improvement function on renal fibrosis and was proportional to the dose.

[0089] according to Fig.11 and Fig.12 It can be seen that serum BUN and CRE in the model group induced by L-NAME for 14 weeks increased significantly. After 8 weeks of administration, serum BUN and CRE in the captopril group decreased significantly. There was no significant difference in serum BUN and CRE in the 15 mg / kg AEL group. Serum BUN and CRE in the 30 mg / kg and 60 mg / kg AEL groups decreased significantly, which was proportional to the dose.

[0090] AEL can reduce the levels of serum BUN and CRE in hypertensive mice, alleviate pathological phenomena such as renal interstitial fibroblast proliferation, inflammatory factor infiltration, basement membrane thickening and volume increase of glomerular capsule wall in hypertensive mice, thereby improving renal function impairment.

[0091] 7. Detection of aortic vascular tissue indicators

[0092] Example 1 Mice were killed 2 hours after the last administration, and blood samples were collected and coagulated at room temperature for 2 hours. The blood samples were centrifuged at 4°C and 3500 rpm / min for 10 minutes to separate the serum for the detection of serum nitric oxide (NO) and endothelin-1 (ET-1). The thoracic aorta was carefully separated along the aortic arch, and the connective tissue was removed. Do not pull to cause artificial vascular damage. The thoracic aorta was frozen and sectioned, and HE staining was performed. The results are shown in Figure 13 to Figure 15 shown.

[0093] according to Figures 13-15 It can be seen that the NO level in the model group decreased significantly and the ET-1 content increased significantly, indicating that long-term hypertension caused endothelial dysfunction; the NO level in the captopril group increased and the ET-1 level decreased, which significantly alleviated the endothelial dysfunction caused by hypertension; AEL intervention can increase the NO level, which is positively correlated with the dose, while there is no significant difference in ET-1 ( Fig.13 ); The vascular smooth muscle cells of the control group were closely arranged, the intima was smooth, and the endothelial cells were intact; the cells of the model group were loosely arranged, the smooth muscle was hypertrophic, the intima was rough, the endothelial cells were swollen and ruptured, the intima-media thickness increased, and the vascular lesions were significant; the intima-media thickness of the captopril group was significantly reduced, and the aortic vascular remodeling was significantly improved; the smooth muscle cells of the medium and high dose AEL groups were closely arranged, and the intima was relatively smooth ( Fig.14 ), the thickness of the intima-media layer was significantly reduced ( Fig.15 ), improved aortic vascular remodeling in a dose-proportional manner. AEL can increase NO content and prevent aortic remodeling caused by hypertension.

[0094] 8. Renin-angiotensin-aldosterone system (RAAS system) detection

[0095] Example 1 Mice were killed 2 hours after the last administration, blood samples were collected, coagulated at room temperature for 2 hours, centrifuged at 4°C, 3500 rpm / min for 10 minutes, serum was separated, and the levels of renin (Renin), angiotensin converting enzyme (ACE), angiotensin II (AngII) and adrenal cortex aldosterone (ALD) were detected. The results showed that the levels of Renin, ACE, AngII and ALD in the serum of the model group increased significantly. After 8 weeks of administration, captopril and AEL could significantly reduce the levels of ACE, AngII and ALD, but had no significant effect on Renin, and the regulatory effect was proportional ( Fig.16 ).

[0096] According to the above content, it can be seen that black wolfberry anthocyanins (AEL) have a hypotensive effect, can protect target organs such as the heart and kidneys, alleviate their organ damage, and reduce TG and LDL levels to reduce the risk of cardiovascular disease, which is proportional to the dose. The pharmacodynamic mechanism of AEL in lowering blood pressure is related to reducing the level of angiotensin converting enzyme (ACE), reducing the secretion of angiotensin II (AngII) and aldosterone (ALD), regulating the over-activation of the RAAS system; increasing the level of nitric oxide (NO), reducing vascular tension; reducing the thickness of the vascular intima-media layer (IMT) to improve aortic remodeling, and has the advantages of multi-link and multi-target overall regulation.

[0097] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of anthocyanins from black wolfberry in the preparation of blood pressure lowering products; The black wolfberry anthocyanidins include petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-O-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside, malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside; The petunidin-3-O-rutin (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanins in the black wolfberry.

2. The use according to claim 1, It is characterized in that The blood pressure lowering products include products for preventing and / or treating cardiovascular diseases.

3. The use according to claim 2, It is characterized in that The cardiovascular disease products include hypertension and / or hyperlipidemia.

4. The use according to claim 2, It is characterized in that The blood pressure lowering products include products that prevent vascular remodeling and / or reduce vascular tone.

5. The use according to claim 4, It is characterized in that The prevention of vascular remodeling includes reducing the thickness of the vascular intima-media layer; The reduction of vascular tone includes increasing vascular nitric oxide.

6. The use according to claim 2, It is characterized in that The blood pressure lowering products include products that improve abnormal lipid metabolism.

7. The use according to claim 6, It is characterized in that The lipid metabolism index includes one or more of total cholesterol, triglycerides, high-density lipoprotein, low-density lipoprotein and arteriosclerosis index.

8. The use according to claim 2, It is characterized in that The blood pressure lowering products include products that improve the oxidative stress response of hepatocytes and / or regulate the overactivation of the renin-angiotensin-aldosterone system.

9. A blood pressure lowering product, It is characterized in that The efficacy or effective ingredient of the blood pressure lowering product is black wolfberry anthocyanidin; The black wolfberry anthocyanidins include petunidin-3-O-rutinoyl (trans-p-coumaric acid-7-O-glucoside)-5-O-glucoside, petunidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside, petunidin-3-O-rutinoyl (feruloyl)-5-O-glucoside, petunidin-3-O-rutinoyl (caffeoyl)-5-O-glucoside, malvidin-3-O-rutinoyl (trans-p-coumaric acid)-5-O-glucoside; The petunidin-3-O-rutin (trans-p-coumaric acid)-5-O-glucoside accounts for 80-90% of the anthocyanins in the black wolfberry; The products include medicines or health foods.

10. The blood pressure lowering product according to claim 9, It is characterized in that The product is formulated into a single-use dosage form with 115 to 462 mg of Lycium barbarum anthocyanins per dose.

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