Tie2 activator, blood vessel maturation agent or blood vessel stabilizer, and food or drink thereof, blood capillary improvement agent, and blood capillary improvement food or drink

Abstract

The present invention provides: a Tie2 activator, a blood vessel maturation agent, or a blood vessel stabilizer, which activates a Tie2 receptor by means of an active ingredient, promotes blood circulation, and increases the absorption rate of the active ingredient in the body, thereby achieving a capillary improvement effect; also provided are: a food or drink containing the preparation; a capillary-improving agent; and a capillary-improving food or drink. A tea drink according to the present invention contains, as active ingredients, a processed product of South African Robos leaves and / or branches, a processed product of Cinnamon Cinnamomum leaves, a processed product of Cinnamon Cinnamomum branches and / or barks, a processed product of Carthamus tinctorius and a processed product of black beans. The tea is provided for 16 volunteers to drink, and a microscope is used for observing and measuring the blood capillary state and the blood flow velocity of fingers before and after drinking so as to detect whether the food or drink has an improvement effect on the blood capillary. Results show that after the tea is drunk, the shape of blood capillaries, the length of blood vessel loops, the width of the blood vessels, the brightness of tissues around the blood capillaries and the blood flow velocity are all improved.

Description

Technical Field

[0001] The present invention relates to a Tie2 activator, a vascular maturation agent or a vascular stabilizer and a food and drink thereof, as well as a capillary improving agent and a capillary improving food and drink. Background Art

[0002] Capillaries are small reticular blood vessels located between arterioles and venules in the peripheral part of blood vessels, with a diameter of about 6 to 14 microns, accounting for 99% of the blood vessels in the body. Capillaries transport nutrients, oxygen, hormones, etc. to body tissues and cells, while recycling carbon dioxide and metabolic waste and excreting them through various excretory organs. Capillaries are also involved in the regulation of water and body temperature. Capillaries are an extremely important part of the human blood circulation system, and their health is closely related to various organs, especially cardiovascular function. Therefore, capillaries have a profound impact on the health of the body.

[0003] Vascular endothelial growth factor (VEGF) is involved in the formation of new capillaries. When VEGF acts on vascular endothelial cells, it induces cell division, migration, and differentiation, leading to the formation of new blood vessels branching from existing blood vessels. On the other hand, the structural maturation of new capillaries and the structural stability of mature capillaries are caused by the binding of angiopoietin-1 (Ang-1) secreted by capillary wall cells and receptor-type protein tyrosine kinase (tyrosine kinase with Ig and EGF homology domain-2: Tie2) present on the vascular endothelial cell membrane. Therefore, the structural stability of capillaries depends on the functional stability of Ang-1 and Tie2 receptors (see references 1 to 3).

[0004] The capillary wall is formed by a parietal cell directly adhering to the outer surface of the lumen of multiple endothelial cells. Specifically, Ang-1 secreted by the parietal cell binds to the Tie2 receptor on the vascular endothelial cell membrane, phosphorylating the Tie2 receptor and activating it. The activated vascular endothelial cells adhere tightly to each other, forming focal adhesions between endothelial cells. Then, the parietal cell adheres to the focal adhesions of the endothelial cells to form a strong capillary wall.

[0005] When the function of parietal cells is weakened due to aging, lifestyle disorders, inflammatory reactions, excessive reactive oxygen species, etc., the secretion of angiopoietin-1 decreases, and Tie2 receptors cannot be activated, resulting in the shedding of parietal cells, the collapse of adhesion plaques between endothelial cells, the impairment of vascular stability, and the inability to maintain the shape of capillaries. Ultimately, the adhesion plaques between capillary endothelial cells are destroyed, the capillaries are twisted and atrophied, red blood cells aggregate in the lumen, the lumen expands, and the elasticity of the blood vessels decreases. Due to the gaps between endothelial cells, cells in the capillaries and internal environmental factors (such as humoral factors, etc.) can easily seep out of the blood vessels, causing the blood flow rate to slow down or stop, thereby causing a variety of cardiovascular diseases and accelerating the aging process of the body.

[0006] Studies have shown that human capillaries begin to decline around the age of 45 (see document 4). Among the subjects (aged 30 to 80) who underwent nail fold capillary examinations conducted by the present inventors, more than 80% were observed to have abnormal vascular leakage such as capillary bending, twisting, atrophy, thickening, formation of bypass vessels, dark brown or pink turbidity of perivascular tissue, and other symptoms of capillary deterioration such as decreased blood flow velocity (see below).

[0007] References

[0008] [Document 1] Suni C et al.: Requisite role of Angiopoietin-1, a ligand for the Tie2 receptor, during embryonic angiogenesis, Cell, vol. 87, pp. 1171-1180, 1996.

[0009] [Reference 2] Fukuhara S et al.: Angiopoietin-1 / Tie2 receptor signaling in vascular quiescence and angiogenesis, Histol Histopathol, p. 25, 2010.

[0010] [Document 3] Shigetomo Fukuhara, Naoki Mochizuki: The blood vessel stabilization mechanism brought about by angiopoietin-1 (Angiopoietin-1 Vascular Stabilizationメカニズム), Drugs and Body (くすりとからだ), Japanese Journal of Pharmacology, Volume 136(1), Pages 26-30, 2010.

[0011] [Document 4] Kajiya K et al.: Structural alterations of the cutaneous vasculature in aged and inphotoaged human skin in vivo, J Dermatol Sci., vol. 61, pp. 206-207, 2011. Summary of the invention

[0012] [Problems to be Solved by the Invention]

[0013] In order to overcome the above problems, the inventors have conducted in-depth research and discovered an effective ingredient that can deal with capillary degeneration caused by aging, lifestyle disorders, inflammatory reactions, excessive reactive oxygen species, etc., which can activate capillary Tie2 receptors. At the same time, they have invented a food and drink made of edible plant raw materials that can activate capillary Tie2 receptors, promote blood circulation, and promote the absorption of active ingredients by the intestines.

[0014] [Methods to solve the problem]

[0015] The inventors found that the extract of the leaves of Okinawa cinnamon (Cinnamomum sieboldii, also known as Okinawa cinnamon) has the effect of activating Tie2 receptors, promoting blood vessel maturation or stabilizing blood vessels, and thus completed the present invention. In addition, the inventors completed the invention of a capillary-improving food and drink based on the following four elements: 1) an edible plant ingredient containing an edible plant ingredient that activates Tie2 receptors (Okinawa cinnamon, Ceylon cinnamon, South China cinnamon, rooibos); 2) an edible plant ingredient that promotes blood circulation (safflower); 3) an edible plant ingredient that increases blood flow (black beans); and 4) lactic acid bacteria that improve intestinal flora and promote absorption of active ingredients.

[0016] The specific means for solving the above-mentioned problems are as follows: [1]

[0018] A Tie2 receptor activator containing Okinawa cinnamon leaf extract as an active ingredient. [2]

[0020] Contains Okinawa cinnamon leaf extract as an active ingredient, a vascular maturation agent or vascular stabilizer having a vascular maturation effect or a vascular stabilization effect based on Tie2 receptor activation. [3]

[0022] A food or beverage for one of the purposes of Tie2 receptor activation, vascular maturation and vascular stabilization. A food or beverage containing one of the Tie2 receptor activator described in [1], the vascular maturation agent described in [2], and the vascular stabilizer described in [2]. [4]

[0024] A capillary improving agent containing processed products of rooibos leaves and / or branches, processed products of Okinawa cinnamon leaves, processed products of cinnamon branches and / or barks, processed products of safflower, and processed products of black beans as active ingredients. [5]

[0026] The processed products of the capillary improving agent described in [4] above are all dry and pulverized products. [6]

[0028] The capillary improving agent described in [4] above further contains lactic acid bacteria. [7]

[0030] A food or drink for improving capillaries, comprising the capillary improving agent described in [4].

[0031] [Effects of the invention]

[0032] The present invention uses effective active ingredients of plants to activate Tie2 receptors, promote blood circulation, and increase the absorption rate of the active ingredients in the body, thereby achieving improvement of capillaries.

[0033] The present invention is briefly described above. The following further elaborates on the details of the present invention by describing a specific embodiment of the present invention (hereinafter referred to as "embodiment") and the accompanying drawings of the embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The changes in the values ​​(capillary diameter) before and after ingestion of the food and drink of the present invention are shown.

[0035] Figure 2 The numerical changes (the length of the vascular loop) before and after taking the food and drink of the present invention are shown.

[0036] Figure 3 The numerical changes (blood flow velocity) before and after ingestion of the food and drink of the present invention are shown.

[0037] Figure 4 This is a photograph of the appearance of blood vessels (capillary image) before ingestion of the food and drink of the present invention.

[0038] Figure 5 These are photographs of the appearance of blood vessels (capillary images) after ingestion of the food and drink of the present invention.

[0039] Figure 6 These are photographs of the appearance of blood vessels (capillary loop length and blood flow velocity) before ingestion of the food and drink of the present invention.

[0040] Figure 7 These are photographs of the appearance of blood vessels (capillary loop length and blood flow velocity) after ingestion of the food and drink of the present invention.

[0041] Figure 8 Showing the improvement effect of the Rooibos tea group and the control group (cinnamon + rooibos) on capillary blood flow velocity.

[0042] Fig. 9 Showing the improvement effect of the Rooibos tea group and the control group (cinnamon + rooibos) on capillary length.

[0043] Fig.10 Showing the improvement effect of the Rooibos tea group and the control group (cinnamon + rooibos) on capillary width.

[0044] Fig.11 Showing the improvement effect of the Rooibos tea group and the control group (cinnamon + rooibos) on the number of capillaries.

[0045] Fig.12 The results show the improvement effect of the Rooibos tea group and the control group (cinnamon + rooibos) on capillary deformation.

[0046] Fig.13 The results show the improvement effect of the Rooibos tea group and the control group (cinnamon + rooibos) on the haze around capillaries.

[0047] Fig.14 Showing the comparison of capillary improvement effect between the Rooibos tea group and the control group (cinnamon + rooibos).

[0048] Fig.15 The observation results for the 60-year-old female of the Leguancha group number 15 are shown.

[0049] Fig.16 Observations for Leguancha group number 11, a 52-year-old male, are shown.

[0050] Fig.17 Observations for the 73-year-old male in Lok Tong Tea Group No. 4 are shown.

[0051] Fig.18 Observations for Leguancha group number 14, a 54-year-old male, are shown.

[0052] Fig.19 This is a graph showing phosphorylated Tie2 protein bands in each sample detected by Western blotting.

[0053] Fig. 20 This is a graph of Tie2 phosphorylation data.

[0054] Fig.21 This is a photo of the first test result of Case 1 in the fourth embodiment.

[0055] Fig. 22 This is a photo of the second test result of Case 1 in the fourth embodiment.

[0056] Fig.23 This is a photo of the third test result of Case 1 in the fourth embodiment.

[0057] Fig.24 This is a photo of the fourth test result of Case 1 in the fourth embodiment.

[0058] Fig.25 This is a photo of the first test result of Case 2 in the fourth embodiment.

[0059] Fig.26 This is a photo of the second test result of Case 2 in the fourth embodiment.

[0060] Fig. 27 This is a photo of the third test result of Case 2 in the fourth embodiment.

[0061] Fig.28 This is a photo of the fourth test result of Case 2 in the fourth embodiment.

[0062] Fig.29 It is a picture of immunoglobulin blot of the detection result of the fifth embodiment.

[0063] Fig.30 It is a data chart of the test results of the fifth embodiment. DETAILED DESCRIPTION

[0064] One or more specific embodiments of the capillary improving agent, capillary improving food and drink, Tie2 receptor activator, vascular maturation agent or vascular stabilizer and food and drink thereof related to the present invention will be described in detail below.

[0065] However, sometimes unnecessary overly detailed descriptions may be omitted. For example, detailed descriptions of well-known matters or repeated descriptions of substantially the same components may be omitted. This is done to avoid unnecessary lengthiness of the following description and to make it easier for relevant technicians to understand.

[0066] In addition, the following description is intended to enable relevant technical personnel to fully understand the present disclosure, but is not intended to limit the subject matter of the present invention.

[0067] In addition, unless otherwise specified, all numerical values ​​such as parameters, reaction conditions and component concentrations used in this specification should be understood to be modified by the term "about" in all instances. Therefore, unless otherwise indicated, the numerical parameters shown in the following specification should be regarded as approximate values ​​that may vary at least depending on specific analytical techniques.

[0068] <Explanation of terms>

[0069] “Comprising…” and “containing…” Synonymous terms such as “comprising…” or “characterized by…” should be interpreted as having a broad or open meaning and do not exclude other unrecited elements or method steps.

[0070] Furthermore, the term “essentially consisting of…” as used in this specification includes not only specific elements or method steps, but also portions that have no substantial effect on the main components and novel features (singular or plural) of the object for which protection is sought.

[0071] Regarding the terms "comprising...", "consisting of..." and "consisting essentially of...", when any of these terms are used in this specification, the objects disclosed and claimed by the present invention may also include the use of the other two terms. Therefore, in some embodiments not explicitly stated, any case of "comprising..." may be replaced by "consisting of..." or "consisting essentially of...".

[0072] The term "process" or "step" may be used explicitly or implicitly in connection with a process or method feature. However, there is no limitation on the order or procedure between these explicit or implicit steps or steps unless the order or procedure is specified.

[0073] <First Embodiment>

[0074] The first embodiment of the present invention will be described below. This embodiment relates to a capillary improving agent and a capillary improving food and drink.

[0075] In this embodiment, the capillary improving food and beverage (hereinafter referred to as "food and drink") contains at least one processed product from the leaves and branches of rooibos, at least one processed product from the leaves and branches of cinnamon, and processed products of safflower and black beans as the active ingredient of the capillary improving agent. The aforementioned processed products are all dried and crushed. In addition, the food and drink of this embodiment also contains lactic acid bacteria.

[0076] In other words, the food and drink of this embodiment is based on the following three elements: containing plant extracts that can activate Tie2 receptors, plant ingredients that promote blood circulation and increase blood volume, and lactic acid bacteria extracts that promote intestinal absorption of active ingredients, mixed with cinnamon plants and rooibos plants that have Tie2 receptor activating ingredients, safflower that promotes blood circulation, black beans that can increase blood flow, and tea made from lactic acid bacteria that improves intestinal flora and thus enhances intestinal absorption. The inventors call this mixed tea that improves capillary function "Leguan Tea" (see below for details).

[0077] The active ingredients of the food and beverage of the present embodiment, cinnamon ( Cinnamomum ) and rooibos ( Aspalathus linearis ) contain a substance that can activate the Tie2 receptor similar to angiopoietin-1.

[0078] In addition, cinnamon has a spicy and sweet taste, and is warm in nature. It can warm the body, act on multiple meridians, dispel cold and improve the circulation of yang energy, thereby activating blood circulation and relieving pain caused by cold, such as abdominal pain, joint pain, menstrual pain, etc. Rooibos is caffeine-free, low in tannins, and contains more polyphenols than green tea.

[0079] Safflower, a herbal medicine commonly used in Chinese medicine, is warm in nature and has a significant effect on improving blood circulation. It can warm the body and improve various symptoms caused by blood circulation disorders. It can help regulate menstrual disorders and cold symptoms by activating blood circulation, eliminating blood stasis, relieving pain, and helping to regulate menstrual disorders and cold symptoms. Due to its effects of promoting blood circulation and warming the body, it is included in the Japanese Pharmacopoeia as a herbal medicine for promoting blood circulation.

[0080] Black beans are reported to be rich in anthocyanins, a functional substance that can increase blood flow. Anthocyanins have a strong antioxidant effect and can inhibit platelet coagulation, so they are expected to protect capillaries and improve vascular and circulatory functions. In addition, anthocyanins have been confirmed to reduce the risk of vascular-related death.

[0081] There are no particular restrictions on the processed products of rooibos, cinnamon, safflower and black beans, and the processed parts can be appropriately selected according to the purpose, for example, leaves, branches, stems, flowers, fruits, roots and skins, etc. For rooibos, leaves and / or branches are preferred; for cinnamon, skins and / or leaves are preferred; for safflower, flowers are preferred; and for black beans, beans and / or skins are preferred.

[0082] The method for preparing the parts of these processed products is to use the desired parts directly or crush them with a pulverizer after drying, and then use the dried and crushed products as tea materials. In this embodiment, the crushed products of each dried product are stored in packaging bags such as tea bags and provided as capillary improvement food and beverages. Drying can be carried out in the sun or with a commonly used dryer.

[0083] The dieter can boil the tea bag or soak it in hot water to extract its effective ingredients, and then drink it. Through this diet, the dieter can take in the effective ingredients that improve capillaries.

[0084] In the present embodiment, although all processed products are dried and crushed products, it is not limited thereto, and extracts (extracts) may also be used. The extracts may be simply obtained by a general method of plant extraction. There are no particular restrictions on the extracts, and they may be appropriately selected according to the purpose, for example, a diluent of the extract, a concentrated solution, a dried product of the extract, etc.

[0085] In addition, the extraction method is not particularly limited and can be appropriately selected according to the purpose. For example, a method of adding the above-mentioned plant extraction part to a treatment tank filled with an extraction solvent, stirring appropriately to dissolve soluble components if necessary, and then removing the extraction residue by filtration to obtain an extract can be used as an example.

[0086] In addition, the food and beverage of this embodiment contains lactic acid bacteria with intestinal regulating effects. EC-12 strain of lactic acid bacteria is the preferred bacteria. EC-12 strain can improve intestinal flora and enhance the digestion and absorption function of the intestine. Studies have shown that after taking EC-12 strain of lactic acid bacteria, the number of bifidobacteria in the intestine increases by about 2.3 times, and the amount of stool also increases (reference: Terada A et al.: Effects of the consumption of heat-killed Enterococcus faecalis EC-12 preparation on microbiota and metabolic activity of the faeces in healthy adults, Microbial Ecology in Health and Disease, Vol. 16 (4), pp. 188-194, 2004). Therefore, this lactic acid strain is added to many health food and beverages or supplements to promote the absorption of active ingredients. In the present invention, lactic acid bacteria are not necessarily included, but in order to promote efficient absorption of the active ingredient, it is preferable to add lactic acid bacteria.

[0087] In addition, the extraction conditions (extraction time and extraction temperature) and the amount of the extraction solvent used are not particularly limited and can be appropriately selected according to the purpose.

[0088] <Second embodiment>

[0089] The second embodiment of the present invention will now be described. This embodiment relates to a Tie2 activator, a vascular maturation agent or a vascular stabilizer, and a food and drink thereof.

[0090] In this embodiment, an extract of leaves (leaf part) of Okinawa cinnamon (scientific name: Cinnamomumsieboldii Meisn.) extracted with alcohol (e.g., methanol, etc.) is used as a Tie2 receptor activator, a vascular maturation agent, or a vascular stabilizer and its food and drink. That is, the extract of leaves of Okinawa cinnamon is used as an active ingredient for activating Tie2 receptors. It is worth noting that Okinawa cinnamon is also called Okinawa Karaki in Okinawa.

[0091] Okinawa cinnamon (scientific name: Cinnamomum sieboldii Meisn. or Cinnamomum Okinawense Hatus., nom. nud., alias: Okinawa Karaki) is a native species of Okinawa. It is only confirmed to be distributed wild in Okinawa in Japan (source: Plants of the World Online, recorded as Cinnamomum sieboldii Meisn.). Those grown in areas outside Okinawa (including Honshu, Japan) are all cultivated varieties. Although this variety is sometimes believed to have been introduced from China, there is no record of this variety in the Chinese tree record, and there is no corresponding variety description in the "Flora of China" (source: Mikawa's Flower (Flora of Mikawa) chapter).

[0092] Okinawan Karaki is a different plant species from the cinnamon (Cinnamomum cassia (L.) D. Don;) most commonly used in China.

[0093] The main differences are as follows:

[0094] 1. Appearance and flowering period:

[0095] Okinawa Karaki is a large evergreen tree 10 to 15 meters tall, with alternate leaves, smooth and glossy surface. It blooms with small yellowish-green flowers in clusters from May to June, and its fruit is an oval berry that turns dark brown when ripe.

[0096] In contrast, cinnamon is a medium-sized tree with alternate or nearly opposite leaves and very fine yellow-brown hairs on the surface. It blooms small yellow-white flowers in three-flowered inflorescences from June to August, and its fruits are oval and dark purple when ripe.

[0097] 2. Aroma and coumarin content:

[0098] Cinnamon has a strong aroma and high coumarin content, and excessive intake may cause liver toxicity or carcinogenic risks (AFC, 2004).

[0099] Okinawa Karaki has a milder aroma and lower coumarin content, making it safer to use.

[0100] Although it has been demonstrated in in vitro test tube experiments that extracts obtained from cinnamon (the bark of the trunk) can activate Tie2 receptors, it is not clear whether its leaves have the effect of activating Tie2 receptors. In addition, since Okinawa cinnamon is a precious tree in Okinawa and has been designated as a quasi-extinct endangered tree species and protection activities have been carried out, the bark of the trunk cannot be used. Therefore, further research and development and application of the leaves of Okinawa cinnamon are of great value. After in-depth research, the inventors found that the leaf extract of Okinawa cinnamon contains active substances that can activate Tie2 receptors. It is also known that the leaves of Okinawa cinnamon are rich in aldehydes, cloves, phenolic compounds, and catechin trimers, which have the effects of dilating peripheral blood vessels and increasing blood flow.

[0101] The leaf extract of Cinnamon bark used in the present embodiment can be easily obtained by a general method of plant extraction. The extract is not particularly limited and can be appropriately selected according to the purpose, such as a diluted extract, a concentrated extract, or a dried extract.

[0102] In addition, the extraction method is not particularly limited and can be appropriately selected according to the purpose. For example, the extraction part of the plant is added to a treatment tank filled with an extraction solvent, and if necessary, the soluble components are appropriately stirred to dissolve, and then the extraction residue is removed by filtration to obtain an extract. In this embodiment, the extraction solvent is an alcohol.

[0103] In addition, the leaf extract or powder (e.g., dried and ground) of Cinnamon bark in this embodiment can be mixed into the food and drink used as the capillary improving agent in the first embodiment. After mixing, the activation of Tie2 receptors, the maturation of blood vessels, or the stabilization of blood vessels can be further enhanced, which has a synergistic effect.

[0104] In addition to the processed leaves of Okinawa cinnamon of the present embodiment, the capillary improving agent or food and drink as an effective ingredient may also include processed leaves and / or branches of rooibos, processed branches and / or bark of other cinnamon, processed safflower, and processed black beans. Such food and drinks will be explained from the perspective of traditional Chinese medicine.

[0105] Chinese medicine has the concept of "activating blood circulation and removing blood stasis". Disorders in lifestyle and other factors lead to the occurrence of "blood stasis", and certain plants or foods are believed to improve blood flow obstruction in blood vessels, improve pain, cold symptoms, skin metabolism and blood circulation, which is the effect of "activating blood circulation and removing blood stasis".

[0106] The present invention is based on this Chinese medicine concept, and the inventor has conducted in-depth research. As a result, the inventor has developed a food and drink that can improve capillaries (tea is one example) by adding the precious plant Okinawa cinnamon mentioned in this embodiment, which has the effect of promoting blood circulation and removing blood stasis, "cassia bark (cinnamon bark)", "safflower" and "black beans" known to the Chinese people since ancient times.

[0107] Chinese medicine attaches importance to the properties (five properties: warm, hot, cool, cold, and neutral), taste (five tastes: sour, bitter, sweet, pungent, and salty), meridians (liver meridian, heart meridian, kidney meridian, spleen meridian, and lung meridian), and yin and yang of plants and foods (references: "Chinese Pharmacopoeia 2020 Edition", "Chinese-Japanese Medical Dictionary (China People's Medical Publishing House)", "Toyama University Chinese and Chinese Medicine Research Institute and Chinese Medicine Database Portal", and "Medicinal Plant Comprehensive Information Database [National Research Institute for Pharmaceutical Research]").

[0108] In the aforementioned first embodiment or second embodiment, cassia bark (cinnamon bark) is considered to have the characteristics of "hot in nature, pungent and sweet in taste, entering the kidney, spleen, heart, and liver meridians, and replenishing yang". Specifically, its effects include replenishing yang, dispelling cold, relieving symptoms such as cold pain in the stomach and abdomen, dysmenorrhea, and frequent urination, and enhancing yang. As mentioned above, a new raw material (Okinawa cinnamon leaf) is introduced in this embodiment, which can promote blood circulation, warm the internal organs, increase blood flow, and has the effects of sweating, relieving fever, and expelling cold.

[0109] Safflower is warm in nature, pungent in taste, and enters the heart and liver meridians, nourishing yin. Specifically, its effects mainly include "activating blood circulation", "dispelling cold" and "regulating menstruation". Black beans are neutral in nature, sweet in taste, enter the kidney and spleen meridians, and nourish yin. Its effects include "activating blood circulation", "diuresis", "removing wind" and "detoxification".

[0110] The yin and yang balance of these plants is harmonized, with cinnamon (Okinawa cinnamon and other cinnamon), safflower and black beans as the core, and South African rooibos tea and lactic acid bacteria that improve intestinal absorption function are added to promote capillary circulation, thereby achieving the effect of promoting blood circulation and removing blood stasis and improving blood flow (which will be explained in detail in the examples described later).

[0111] The dosage form is not limited to tea. The vascular improver of the present invention can be provided in various forms. Liquid forms include syrups, elixirs, tinctures, emulsions, etc. Solid forms can be tablets, capsules, granules, powders, etc. In powder form, it can be soluble powder and granular powder. In addition, percutaneous absorption preparations, sprays, gels and other dosage forms can also be used. These various dosage forms can be selected according to the user's preferences and circumstances to achieve effective intake.

[0112] In order to explain the usefulness of the present invention in more detail, a number of administration tests are listed below as examples (application examples and / or specific examples) of the present invention. It should be noted that the present invention is not limited to these examples.

[0113] <First embodiment>

[0114] In this example, in order to verify the effectiveness of the food and drink involved in the present invention (i.e., the mixed tea in the first embodiment) in improving capillaries, a human drinking confirmation test was conducted. Figures 1 to 7 , analyze the effectiveness of the food and drink involved in the present invention.

[0115] Figure 1 This is a graph showing changes in values ​​before and after food intake (capillary diameter).

[0116] Figure 2 The graph shows the changes in values ​​before and after food intake (capillary loop length).

[0117] Figure 3 This is a graph showing changes in values ​​before and after food intake (blood flow velocity).

[0118] Figure 4 This is a photograph of the blood vessel condition before taking food or drink (capillary image).

[0119] Figure 5 This is a photograph of the blood vessel condition after ingesting food and drink (capillary image).

[0120] Figure 6 This is a photograph of the blood vessel condition before ingestion of food and drink (length of capillary loop and blood flow velocity).

[0121] Figure 7 These are photos of blood vessel conditions after ingestion of food and drink (capillary loop length and blood flow velocity).

[0122] [Objects and methods]

[0123] The research subjects were 5 volunteers (aged 50 to 73 years old, 1 male and 4 females), each of whom drank the food and drink of the present invention (one package of 3 g) every day by soaking it in about 150 ml of boiling hot water for 5 minutes and then drinking it.

[0124] Using a capillary microscope (manufactured by GOKO Imaging Equipment Co., Ltd., Japan) equipped with high-function image analysis and measurement software GOKO Measure Plus and dedicated flow rate measurement software GOKO-VIP, the nail fold capillaries of the ring finger of the non-dominant hand were observed and measured twice, before drinking the food and beverage of the present invention and 30 days after drinking.

[0125] The parameters measured include: (1) capillary shape, (2) brightness of tissue surrounding the capillaries, (3) length of the vascular loop, (4) thickness of the blood vessels, and (5) blood flow velocity. The measured values ​​of the vascular loop length, blood vessel thickness, and blood flow velocity of each subject are averaged three times at the same location. The p-value probability of the difference between the two average values ​​before and after drinking the food and beverage of the present invention is calculated using a paired sample t-test.

[0126] [result]

[0127] The results of observation and measurement are shown in Table 1 and Figures 1 to 7 .

[0128] Table 1

[0129]

[0130] Table 1 lists the measured values ​​of various parameters of capillaries before and after drinking the food and drink of the present invention.

[0131] (1) Regarding the shape of blood vessels, the ideal fingertip capillary loop should be straight, without lateral bypass, and the vascular loop should not be twisted or deformed.

[0132] (2) The brightness of the tissue around capillaries is affected by factors such as vascular aging, inflammation, and vascular leakage. Waste, vascular leakage substances, and inflammatory response substances caused by these factors will accumulate in the tissue, forming a thick mist.

[0133] (3) The ideal thickness of the capillaries in the nail folds of the fingertips is about 6 to 14 μm. Before drinking the food and drink of the present invention, the average thickness of the capillaries was 16.4±2.5 μm, and after drinking it, the average thickness became 14.3±3.0 μm.

[0134] (4) The ideal length of the capillary loop of the fingertips is more than 250 μm. The average length of the capillary loop before drinking was 259.2 ± 58.1 μm, and the average value after drinking was 321.3 ± 141.4 μm.

[0135] (5) The faster the capillary blood flow velocity, the better. The average value of the capillary blood flow velocity before drinking the food and drink of the present invention was 204.2±166.4 μm / s, and the average value after drinking was 459.1±311.6 μm / s.

[0136] pass Figures 1 to 3 , indicating the changes in capillary thickness, vascular loop length and blood flow velocity before and after drinking the food and drink of the present invention (*, p < 0.05).

[0137] Regarding the thickness of blood vessels, the average value after drinking was 2.1 μm less than before drinking (p < 0.046, with a significant difference). The average value of vascular loop length increased by 62.1 μm compared to before drinking, but the difference was not significant (p > 0.11). The average value of blood flow velocity increased by 254.9 μm / s compared to before drinking (p < 0.018, with a significant difference).

[0138] Figure 4 and Figure 5 The capillary images (145 times magnification) of a 61-year-old female volunteer before and after drinking the food and drink of the present invention for 30 days (3 g per day) are shown.

[0139] like Figure 4 and Figure 5 As shown in the figure, before drinking, there was a thick brown mist in the tissue around the capillaries, making the capillaries unclear. After drinking, the brown mist disappeared, the capillaries were clearly visible, and the capillary loops were extended.

[0140] Figure 6 and Figure 7 The graph shows the changes in capillary loop length and blood flow velocity (560 times) of a 61-year-old female volunteer before and after drinking the food and drink of the present invention for 30 days (3 g per day).

[0141] like Figure 6 and Figure 7 As shown in the figure, the length of the capillary loop marked 1 in the picture increased from 201.3 μm before drinking to 296.2 μm after drinking, an increase of 1.5 times. The blood flow velocity increased from 286.6 μm / s before drinking to 424.8 μm / s, an increase of 1.6 times.

[0142] Figure 6 and Figure 7The length of the blood vessel ring marked 2 in the picture increased by 1.7 times from 331.9 μm before drinking to 561.3 μm after drinking. The blood flow velocity increased by 1.8 times from 238.8 μm / s before drinking to 411.1 μm / s.

[0143] Figure 6 and Figure 7 The blood flow velocity of the blood vessel marked 3 in the picture increased from 50.7 μm / s before drinking to 331.9 μm / s, which is 3.8 times faster. In addition, the capillary loops marked 1 and 2 were curved before drinking, but became straight after drinking.

[0144] [Investigation and Conclusion]

[0145] In this experiment, after drinking the food (mixed tea) containing cinnamon, rooibos, safflower, black beans and EC-12 lactic acid bacteria for 30 days, the morphology of capillaries, the brightness of the tissue around the capillaries, the thickness of capillaries, the length of capillary loops and the blood flow rate of capillaries were all improved. In particular, the thickness of capillaries and the blood flow rate were significantly improved, which confirmed that the present invention has the effect of improving the health of human capillaries.

[0146] <Second embodiment>

[0147] In this example, in order to verify the effectiveness of the food of the present invention (i.e., the mixed tea in the first embodiment) in improving the capillary function of the fingernail wrinkles, a test was conducted on subjects drinking the mixed tea. In addition, the mixed tea in this example is also called "Leguan Tea" (see below for details).

[0148] [Purpose]

[0149] In the present embodiment, among the subjects whose fingertip nail fold capillaries were examined in the age group of 30 to 80 years old, more than 80% showed vascular deformation phenomena such as vessel bending, twisting, atrophy, thickening, bypass vessel formation, as well as dark brown (accumulation of metabolic products) or pink (vascular leakage) mist-like abnormal phenomena, and vascular degeneration symptoms such as decreased blood flow velocity (see below for details).

[0150] In vitro test tube experiments have shown that substances extracted from cinnamon, rooibos and pepper have the effect of activating Tie2, but due to factors such as intestinal absorption and blood flow status, the effect on the capillaries in the body after taking them is not yet clear. In order to solve the capillary degeneration caused by factors such as aging, lifestyle disorders, inflammation, and excessive reactive oxygen, the inventors have formulated a mixed tea food called "Leguan Tea" containing cinnamon and rooibos that activate Tie2, safflower that promotes blood circulation, black beans that increase blood flow, and lactic acid bacteria that improve intestinal flora and enhance intestinal digestion and absorption functions, based on the three major elements of ingredients that activate Tie2 receptors, ingredients that promote blood circulation, and ingredients that promote the absorption of effective ingredients in the intestine, and verified the effect of improving capillaries through in vivo experiments.

[0151] [Subjects, Materials, and Methods]

[0152] [Target]

[0153] Thirty-two volunteers ranging in age from 38 to 81 years old (average age 57, 18 males and 14 females) were randomly divided into two groups: "Leguan Tea Verification Group" and "Cinnamon + Rooibos Control Group", with 16 people in each group.

[0154] [Material]

[0155] [1] The validation group drank "Leguan Tea" (containing Okinawa cinnamon leaf crushed and powdered, Ceylon cinnamon or South China cinnamon bark crushed and powdered, Rooibos branch and leaf crushed and powdered, safflower crushed and powdered, black bean crushed and powdered, and lactic acid bacteria extract).

[0156] [2] The control group drank “Cinnamon + Rooibos” (a mixture of cinnamon and rooibos with the same content as in Lopix tea).

[0157] [method]

[0158] [1] The validation group and control group drank “Leguan tea” (3 g / day) and “Cinnamon + Rooibos” tea (2 g / day), respectively, soaked in about 150 ml of boiling water for at least 5 minutes, and drank for 30 consecutive days.

[0159] [2] Using a capillary microscope (manufactured by GOKO Imaging Equipment Co., Ltd., Japan) equipped with high-performance image analysis and measurement software GOKO Measure Plus and GOKO Bscan-Z / ZD dedicated flow velocity measurement software GOKO-VIP, the capillaries in the nail fold of the ring finger of the non-dominant hand were observed and measured one day before and 30 days after drinking "Loco Tea" and "Cinnamon + Rooibos".

[0160] [3] Precautions during measurement: The room temperature should be maintained at 26±1℃, the subject should be seated, the measuring finger should be kept at the same height as the heart, and the same part should be observed and measured twice.

[0161] [4]Measurement and observation parameters (variables):

[0162] (1) Capillary morphology (scores for deformed vessels such as twisting):

[0163] In one field of view (145x), according to the proportion of deformed blood vessels, <10% = 0; >10%, <30% = 1; >30%, <60% = 2; >60% = 3. The ideal fingertip capillary loop should be straight, without lateral bypass, and the capillary loop should not be distorted.

[0164] (2) Brightness of tissue surrounding capillaries (fog-like shadow):

[0165] According to the proportion of fog in a field of view (145 times), none = 0; 1 / 3 of the field of view = 1; 1 / 2 of the field of view = 2; > 1 / 2 of the field of view = 3. The brightness of the tissue around the capillaries is affected by factors such as vascular aging, inflammation and vascular leakage. These factors cause waste or vascular leakage substances and inflammatory reactants to accumulate in the tissue, forming a thick fog.

[0166] (3) Length of vascular loop:

[0167] Average length of the five longest vascular loops at 145x magnification (μm). The ideal length of the fingertip capillary loop should be more than 250μm.

[0168] (4) The thickness of blood vessels:

[0169] The average diameter of five blood vessels at 145x or 560x magnification (μm). The ideal thickness of fingertip capillaries should be about 6 to 14 μm.

[0170] (5) Number of capillaries:

[0171] The number of capillary loops in the first row in one field of view at 145x magnification. The ideal number of capillaries in a fingertip should be >20.

[0172] (6) Blood flow velocity:

[0173] The average blood flow velocity (μm / s) of the arterial side of the three capillary loops was calculated from the video. The faster the capillary blood flow velocity, the better.

[0174] [5] Statistical analysis:

[0175] Data are presented as mean ± SEM of the values ​​obtained in each group (n = 16). Differences in mean values ​​were confirmed by ANOVA procedure and bivariate t-test analysis without replication. The significance threshold was set as the value divided by the number of comparisons (P < 0.05).

[0176] [Test results]

[0177] The test results are shown in Tables 2 and 3. Table 2 shows the original data and subject list of the Leguancha group. Table 3 shows the original data and subject list of the control group.

[0178] Table 2

[0179]

[0180] Table 3

[0181]

[0182] [Statistical Analysis]

[0183] The test results were statistically analyzed. The analysis results are shown in Table 4, Table 5 and Figures 8 to 14 shown.

[0184] Table 4 shows the effects of drinking “Leguan Tea” and “Cinnamon + Rooibos” on capillaries.

[0185] Table 5 compares the capillary improvement effects of “Leguan Tea” and “Cinnamon + Rooibos”.

[0186] Figure 8 The improvement effect of Rooibos tea and the control group (cinnamon + rooibos) on capillary blood flow velocity was shown.

[0187] Fig. 9 The improvement effect of Rooibos tea on capillary length was shown compared to the control group (cinnamon + rooibos).

[0188] Fig.10 The results show the improvement effect of Rooibos tea and the control group (cinnamon + rooibos) on the thickness of capillaries.

[0189] Fig.11 The improvement effect of Rooibos tea and the control group (cinnamon + rooibos) on the number of capillaries was shown.

[0190] Fig.12 The improvement effect of Leguan tea and the control group (cinnamon + rooibos) on deformed capillaries was shown.

[0191] Fig.13 The improvement effect of Leguan tea and the control group (cinnamon + rooibos) on pericapillary turbidity is shown. Fig.14Shown is a comparison of the capillary improvement effects of the Rooibos group and the control group (cinnamon + rooibos).

[0192] Table 4

[0193]

[0194] Table 5

[0195] Test items Cinnamon + Rooibos Leguan Tea Average increase in vessel length (μm) 20.1±19.4 <![CDATA[40.1±24.3 * ]]> Average reduction in blood vessel diameter (μm) 0.6±0.5 <![CDATA[1.3±0.9 * ]]> Average increase in the number of blood vessels (line) 1±1 <![CDATA[2±1.7 * ]]>

[0196] As shown in Table 4, Leguan tea showed significant improvement effects in all measurement items, while the cinnamon + rooibos control group only showed weak improvement effects in the length, thickness and number of blood vessels. ***, P < 0.001; **, P < 0.01; *, P < 0.05.

[0197] As shown in Table 5, there were significant differences between the Leguan tea group and the cinnamon + rooibos control group in the mean increase or decrease values ​​of capillary length, thickness, and number of blood vessels. *, P < 0.05.

[0198] like Figure 8 As shown, blood flow velocity increased in both groups, but there was no significant difference in the control group before and after drinking.

[0199] like Fig. 9 As shown, both groups had improved effects. **, P < 0.01; ***, P < 0.001.

[0200] like Fig.10 As shown, both groups had an improvement in capillary thinning. *, P < 0.05.

[0201] like Fig.11 As shown in Figure 2, the improvement effect of both groups was significant. *, P < 0.05; **, P < 0.01. Fig.12 As shown, there was no significant difference in the control group before and after drinking. *, P < 0.05.

[0202] like Fig.13 As shown, the control group showed an improvement trend, but there was no significant difference. **, P < 0.001.

[0203] like Fig.14 As shown, in terms of capillary length (A), thickness (B) and number of blood vessels (C), both groups showed improvement effects, but the difference between the two groups was significant, indicating that the effect of Leguan Tea was significantly better than that of the control group. *, P < 0.05.

[0204] [About the first case of this embodiment]

[0205] Reference Fig.15, the first case observed in the above confirmation test is explained.

[0206] Fig.15 The results of observation for a 60-year-old female of Leguancha group No. 15 (see Table 2) are shown.

[0207] Fig.15 Shown are improvements in capillary blood flow velocity, vascular loop length, perivascular clarity, and vascular loop shape before (left) and after (right) drinking Leguan tea for 30 days (3 g daily).

[0208] [Image above (magnification = 145)]

[0209] Before drinking, thick brown and pink haze (inflammation, vascular leakage and poor metabolism) was visible, and the blood vessels were short and unclear; after drinking, the brown and pink haze was significantly reduced, and the blood vessels became longer and clearly visible.

[0210] [Image below (magnification=560)]

[0211] (1) Blood flow velocity before and after drinking: 1 = 286.6 μm / s → 424.8 μm / s; 2 = 238.9 μm / s → 411.1 μm / s; 3 = 50.75 μm / s → 196.58 μm / s; 4 = 184.3 μm / s → 295.7 μm / s, increasing by 1.6, 1.8, 3.8 and 1.4 times respectively.

[0212] (2) The length of the vascular loop before and after drinking: 1 = 201.3 μm → 296.2 μm; 2 = 181.4 μm → 253.7 μm; 3 = 159.1 μm → 277.2 μm; 4 = 212.8 μm → 394.9 μm, which increased by 1.5, 1.4, 1.7 and 1.9 times respectively.

[0213] (3) Compared with before drinking, the pink haze around the blood vessels decreased after drinking, and the vascular leakage was improved.

[0214] (4) After drinking the drink, the deformed blood vessel loops 1 to 4 became straightened and their shapes improved.

[0215] [About the second case of this embodiment]

[0216] Reference Fig.16 , the second case observed in the above confirmation test is explained.

[0217] Fig.16 The results of observations for a 52-year-old male of Leguancha group No. 11 (see Table 2) are shown.

[0218] Fig.16Shown are the improvements in capillary blood flow velocity, vascular loop length, and blood vessel thickness before (left) and after (right) drinking Leguan tea for 30 days (3 grams per day).

[0219] [Image above (magnification = 145)]

[0220] Before drinking, the blood vessels are short and thick, but after drinking, they become long, thin, and clearly visible.

[0221] [Image below (magnification=560)]

[0222] (1) The average blood flow velocity increased from 107.6 μm / s to 706.5 μm / s before and after drinking, an increase of 6.5 times. (2) The average length of the vascular loop increased from 103.7 μm to 208.8 μm before and after drinking, an increase of 2 times.

[0223] (3) The average thickness of blood vessels decreased from 13.5 μm to 9.3 μm before and after drinking, a decrease of 4.2 μm.

[0224] “↓” marks the same position.

[0225] [About the third case of this embodiment]

[0226] Reference Fig.17 , the third case observed in the above confirmation test is explained.

[0227] Fig.17 The results of observations for a 73-year-old male of Leguancha Group No. 4 (see Table 2) are shown.

[0228] Fig.17 The capillary blood flow velocity, vessel length, vessel thickness, deformed vessels, and improvement of perivascular clarity before (left) and after (right) drinking Leguan Tea for 30 days (3 grams per day) (magnification = 145). Before drinking, the vessels were short and thick, the vascular loops were twisted, and the surroundings were covered with dense pink haze. After drinking, the vessels became longer and thinner, the vascular loops became straighter, and the pink haze was significantly lighter.

[0229] (1) The average blood flow velocity increased from 194.1 μm / s to 489.7 μm / s before and after drinking, a 2.5-fold increase.

[0230] (2) The average length of the vascular ring increased from 175.2 μm to 283.7 μm before and after drinking, a 1.6-fold increase.

[0231] (3) The average thickness of blood vessels decreased from 18.6 μm to 12.3 μm before and after drinking, a decrease of 5.3 μm.

[0232] (4) The proportion of twisted blood vessels (deformed blood vessels) dropped from 60% before drinking to 10% after drinking, and the degree of deformation was significantly reduced.

[0233] “↓” marks the same position.

[0234] [About the fourth example of this embodiment]

[0235] Reference Fig.18 , the fourth case observed in the above confirmation test is described.

[0236] Fig.18 The results of observations for a 54-year-old male of Leguancha group No. 14 (see Table 2) are shown.

[0237] Fig.18 Shown are the improvements in capillary blood flow velocity, blood vessel thickness, and deformed blood vessels before (left) and after (right) drinking Leguan tea for 30 days (3 grams per day) (magnification = 560).

[0238] (1) The average blood flow velocity before and after drinking increased from 238.5 μm / s to 360.9 μm / s, an increase of 1.5 times.

[0239] (2) The average thickness of blood vessels decreased from 12.6 μm to 9.9 μm before and after drinking, a decrease of 2.7 μm.

[0240] (3) The vascular loops that were stuck together before drinking (deformed vascular loops) returned to normal vascular ring shapes after drinking. The proportion of deformed blood vessels dropped from 70% before drinking to 20% after drinking.

[0241] The black “↓” indicates an adhesion-deformed vascular ring, and the white “↓” indicates a normal vascular ring.

[0242] Feedback from Leguan Tea drinkers: Many drinkers say that the tea tastes good and helps improve cold intolerance. Other feedback includes lowered blood pressure, increased appetite, improved constipation, improved complexion, easier sleep, increased morning alertness, and an overall better feeling.

[0243] [Investigation and Conclusion]

[0244] Ingredients in "Loco-Tea", such as cinnamon (Cinnamomum) and South African rooibos (Aspalathus linearis), contain substances that can activate Tie2 receptors, such as s-Syringaresinol, which is similar to angiopoietin-1. In vitro experiments have shown that these ingredients can bind to the Tie2 receptors of vascular endothelial cells in vitro and promote their phosphorylation (activation). In addition, cinnamon has a spicy and sweet taste and warming properties, which can improve blood circulation in the body and relieve symptoms such as abdominal pain, joint pain, and menstrual pain caused by cold. South African rooibos is a plant that does not contain caffeine, has low tannins, and is richer in polyphenols than green tea.

[0245] Safflower has a warming property and can warm the body, promote blood circulation and remove blood stasis, improve various symptoms caused by blood circulation disorders, and promote blood circulation to eliminate blood stasis. Due to its excellent effect in promoting blood circulation, safflower has been included in the Japanese Pharmacopoeia as a raw medicinal material for promoting blood circulation.

[0246] Black beans are rich in anthocyanins, a functional substance that helps hematopoiesis and promotes blood flow. Anthocyanins have strong antioxidant effects, can inhibit platelet aggregation, help protect capillaries, and improve vascular function and circulation. In addition, the use of anthocyanins has been shown to reduce the risk of vascular-related death. Studies have shown that black bean polyphenols can promote the production of NO in vascular endothelial cells, suggesting that it can enhance blood flow.

[0247] EC-12 lactic acid bacteria, which has a bowel-regulating effect, has been proven to improve intestinal flora and enhance digestion and absorption functions. Experiments have shown that the intake of EC-12 lactic acid bacteria can increase the number of bifidobacteria in the intestines to about 2.3 times and increase the amount of bowel movements. This strain has been added to a variety of health products to promote the absorption of active ingredients.

[0248] Studies have shown that cinnamon and rooibos can activate the Tie2 receptors of capillary endothelial cells in vitro, but the improvement of capillaries in vivo is affected by factors such as intestinal absorption and blood flow. It can be inferred that adding ingredients that promote the absorption of active ingredients and promote blood circulation can help enhance their effects in vivo.

[0249] The "Le Tube Tea" in this embodiment contains the above-mentioned ingredients that have the effect of improving capillaries, namely cinnamon, rooibos, safflower, black beans and EC-12 lactic acid bacteria. This study showed that there were significant differences in the six measurement parameters before and after drinking, indicating that "Le Tube Tea" has an improvement effect on capillaries. However, "Cinnamon + Rooibos" only slightly improved the two parameters of blood vessel length and blood vessel thickness. Moreover, in these two parameters, "Le Tube Tea" was significantly higher than "Cinnamon + South African Red Bush" in terms of the average increase in length and number or the decrease in thickness (P<0.05).

[0250] in conclusion:

[0251] (1) "Leguan Tea" contains ingredients that activate Tie2 receptors and promote absorption and blood circulation. It can improve the shape of capillaries, the blurriness of surrounding tissues, thickness, length, number and blood flow rate, and has a positive effect on the health of human capillaries.

[0252] (2) For the beneficial capillary components verified in vitro to work in vivo, they need to involve components that promote intestinal absorption and blood circulation.

[0253] Adding an extract or powder (such as a dry powder) of "Okinawa cinnamon (scientific name Cinnamomum sieboldii Meisn.)" leaves to "Lokuan tea" can further enhance the activation effect of Tie2 receptors.

[0254] <Third Embodiment>

[0255] In this example, Okinawa cinnamon leaves are used, which are rich in aldehydes, eugenol, polyphenols and catechin trimers, and have the effects of dilating peripheral blood vessels, increasing blood flow, and relieving fever and warming. This example verifies the Tie2 receptor activation effect of Okinawa cinnamon leaves.

[0256] [Materials and Methods]

[0257] [Material]

[0258] Dried powder of Okinawa cinnamon (scientific name Cinnamomum sieboldii Meisn.) leaves.

[0259] [Extraction method]

[0260] Extraction with methanol: Specifically, 3 g of leaf powder was added to 60 L of methanol, and the mixture was allowed to stand at 50°C for 3 hours, and the filtrate was concentrated under reduced pressure by suction filtration.

[0261] [Analysis method]

[0262] Tie2 phosphorylation analysis was performed using mouse pro-B cells (Ba / F3-human Tie2) that express a large amount of human Tie2 receptors. Ba / F3-human Tie2 cells were stimulated with angiopoietin-1 (Ang1 300ng / ml), negative control PBS or DMSO, and the test substances. Different concentrations of the test substances were directly added to regular culture medium (10% FBS RPMI1640 + 1pg / ml mouse IL-3), and the cells were washed with PBS and extracted after 15 minutes. The extract was electrophoresed on a 7.5% SDS gel and transferred to a nylon membrane. The membrane was blocked with % skim milk + 0.5% BSA / TBST for 60 minutes and then detected with anti-Tie2 antibody (Ab33, Upstate) and anti-phospho-Tie2 antibody (Tyr992, Cell Signaling Technology). It was then detected with an HRP-labeled secondary antibody and developed with an ECL solution. The phosphorylation degree of Tie2 was measured using Amersham Imager 680 software.

[0263] [result]

[0264] Fig.19 , Fig. 20 Table 6 shows the experimental results.

[0265] Fig.19 The phosphorylated bands of Tie2 protein detected in each sample are shown.

[0266] Fig. 20 is a graph of the Tie2 phosphorylation data in Table 6.

[0267] Table 6 shows the values ​​calculated by Western blotting.

[0268] Table 6

[0269] Tie2 PTie2 PTie2 / Tie2 ①DMSO (10ul / mL) 818332 99361 0.121418935 1 ②Ang1(300ng / mL)+DMSO(10ul / mL) 721526 393722 0.545679574 4.494188 ③B1 (500ug / ml) 754543 85571 0.113407718 0.93402 ④B1 (1000ug / ml) 670025 86646 0.129317563 1.065053 ⑤B2 (500ug / ml) 585234 97052 0.165834521 1.365804 ⑥B2 (1000ug / ml) 599628 130991 0.218453775 1.799174 ⑦B3 (125ug / ml) 570912 92379 0.161809526 1.332655 ⑧B3 (250ug / ml) 576289 69836 0.121182254 0.998051 ⑨A282=2.1(500ug / ml) 555010 74168 0.133633628 1.100600 ⑩A282=2.1(1000ug / ml) 657048 48397 0.073658241 0.606645

[0270] like Fig.19 As shown, the intensity of the detected band is proportional to the phosphorylation amount of Tie2 protein. The higher the band intensity, the more phosphorylated Tie2 protein is.

[0271] (1) DMSO control; (2) Ang1 positive control; (5) Cinnamon leaf extract (500 μg / mL); (6) Cinnamon leaf extract (1000 μg / mL). The intensity of the Tie2 phosphorylation band of (6) was significantly higher than that of (1) DMSO control, and the Tie2 phosphorylation rate was 1.8 times that of the DMSO control (the standard for phosphorylation determination was: higher than 1.5 times).

[0272] As shown in Table 6, (6) the amount of Tie2 phosphorylation in the Okinawa cinnamon leaf extract (1000 μg / mL) was 130991, while that in the DMSO control was 99361, and the phosphorylation rate was 1.8 times higher.

[0273] like Fig. 20 As shown, (6) the Tie2 phosphorylation rate of the Okinawa cinnamon leaf extract (1000 μg / mL) was 1.8 times that of the DMSO control.

[0274] [Investigation]

[0275] Compared with the DMSO control, Tie2 was phosphorylated under the action of Ang1, proving that the analysis method is effective.

[0276] The phosphorylation rate of Tie2 in Okinawa cinnamon leaves (1000 μg / mL) was 1.8 times that of the DMSO control, and phosphorylation of Tie2 was observed. The phosphorylation of Tie2 was also observed by direct visual observation of the band intensity in immuno-western blotting, which verified the effectiveness of the present invention.

[0277] This embodiment uses alcohol extraction, but is not limited thereto. Processing Okinawa cinnamon leaves into powder and drinking with boiled water can also achieve similar effects.

[0278] <Fourth Embodiment>

[0279] In this embodiment, two examples of the improvement effect of using the Leguan tea of ​​the present invention on the capillaries of patients infected with the new coronavirus were verified.

[0280] [Purpose]

[0281] The symptoms caused by the new coronavirus (SARS-CoV-2) infection and the types of organs affected are very wide. The spike protein of SARS-CoV-2 can induce anti-fibrinolytic fibrin, leading to the formation of microthrombi in capillaries. Recent research reports show that the SARS-CoV-2 spike protein forms anti-fibrinolytic fibrin, leading to microthrombi in capillaries (Lize M. et al.: SARS-CoV-2 spike protein S1 induces fibrin(ogen)resistant tofibrinolysis: implications for microclot formation in COVID-19, Biosci Rep. Journal, 41:8, August 27, 2021). When microthrombi block capillaries, blood vessels are deformed, affecting the circulation of blood and oxygen, leading to a variety of symptoms. The present invention aims to confirm whether Leguan Tea is also effective in improving capillaries in patients infected with the new coronavirus.

[0282] [method]

[0283] Using the GOKO-VIP capillary microscope (manufactured by GOKO Imaging Equipment Co., Ltd., Japan) with GOKO Measure Plus high-function image analysis measurement software and GOKO Bscan-Z / ZD dedicated flow measurement software, the experiment was conducted on two patients infected with the new coronavirus. The changes in the nail fold capillaries (vessel length, vessel thickness, blood flow velocity, haze, etc.) of the ring finger of the non-dominant hand were measured multiple times before and after infection and before and after drinking Leguan Tea (3 grams per pack, one pack per day, soaked in about 150 ml of boiling water for more than 5 minutes and then drunk) to study the impact of the new coronavirus infection on capillaries and the effect of Leguan Tea.

[0284] [Results of Case 1]

[0285] The subject of this case is a 63-year-old woman. Figure 21 to Figure 24 The arrow “↓” in the figure indicates the same position.

[0286] Fig.21 Shows the results of the first inspection of Case 1. Photographed on July 4, 2023 (first inspection, magnification 145x).

[0287] Fig. 22 Shows the results of the second inspection of Case 1. Photographed on August 20, 2023 (second inspection, magnification 145x).

[0288] Fig.23 Shows the results of the third inspection of Case 1. Photographed on September 21, 2023 (third inspection, magnification 145x).

[0289] Fig.24 Shows the results of the fourth inspection of Case 1. Photographed on October 26, 2023 (fourth inspection, magnification 145x).

[0290] like Fig.21 As shown, the blood vessel length was observed to be 147.2 microns, the blood vessel width was 13.4 microns, the blood flow velocity was 190.7 microns / second, and the foggy shadow was thicker. In addition, the subject drank the "Cinnamon + Rooibos" mixed tea consumed by the control group in the above-mentioned second embodiment from July 4 to July 17, 2023, and was diagnosed with new coronavirus infection on July 18, 2023, and then stopped drinking it.

[0291] like Fig. 22 As shown, the blood vessel length was observed to be 103.7 microns (shorter than the first 147.2 microns), the blood vessel width was 14.3 microns (thicker than the first 13.4 microns), and the blood flow rate was 101.8 microns / second (significantly slower than the first 190.7 microns / second). In addition, the new coronavirus infection caused significant changes in capillaries. The subjects began to drink Leguan tea from August 20 to September 20, 2023.

[0292] like Fig.23 As shown, the blood vessel length was observed to be 152.2 microns (increased from 103.7 microns in the second time), the blood vessel width was 13.3 microns (thinner than 14.3 microns in the second time), and the blood flow velocity was 215.9 microns / second (a significant increase from 101.8 microns / second in the second time). The subjects continued to drink Leguan Tea from September 21 to October 24, 2023.

[0293] like Fig.24 As shown, the blood vessel length increased to 237.3 microns (longer than before infection), the blood vessel width decreased to 12.1 microns (thinner than before infection), and the blood flow rate reached 406.3 microns / second (significantly faster than before infection). The haze around the capillaries was observed, which was significantly reduced compared with the previous three times, indicating that metabolism has improved.

[0294] Thus, in Case 1, after drinking Leguan tea, the capillaries recovered from the effects of the new coronavirus infection and were even better than before the infection.

[0295] [Results of Case 2]

[0296] The subject of this case is a 69-year-old male. Figure 25 to Figure 28 The arrow “↓” in the figure indicates the same position.

[0297] Fig.25 Shows the results of the first inspection of Case 2. Photographed on July 17, 2023 (first inspection, magnification 145x).

[0298] Fig.26 Shows the results of the second inspection of Case 2. Photographed on August 18, 2023 (second inspection, magnification 145x).

[0299] Fig. 27 Shows the results of the third inspection of Case 2. Photographed on September 24, 2023 (third inspection, magnification 145x).

[0300] Fig.28 Shows the results of the fourth inspection of Case 2. Photographed on October 26, 2023 (fourth inspection, magnification 145x).

[0301] like Fig.25 As shown, the blood vessel length was observed to be 304.6 microns, the blood vessel width was 9.6 microns, and the blood flow velocity was 260.9 microns / second. In addition, the subjects drank the "cinnamon + rooibos" mixed tea consumed by the control group in the second embodiment from July 17 to August 17, 2023.

[0302] like Fig.26 As shown, the blood vessel length was observed to be 282.4 microns, the blood vessel width was 9.3 microns, and the blood flow velocity was 272.9 microns / second, which was not much different from the first examination results. The subject drank Leguan tea for three days from August 18 to August 20, 2023, and was diagnosed with new coronavirus infection on August 21, 2023, and stopped drinking Leguan tea at the same time.

[0303] like Fig. 27 As shown, the vessel length was observed to be shortened to 108.5 μm (significantly shorter than 304.6 μm in the first and 282.4 μm in the second), the vessel width increased to 11.7 μm (larger than 9.6 μm in the first and 9.3 μm in the second), and the blood flow velocity decreased to 129.3 μm / s (significantly slower than 260.9 μm / s in the first and 272.9 μm / s in the second). In addition, the new coronavirus infection caused significant changes in capillaries. The subjects resumed drinking Leguan tea from September 25 to October 26, 2023.

[0304] It was observed that the blood vessel length reached 352.7 microns (longer than before infection), the blood vessel width recovered to 9.6 microns, and the blood flow velocity reached 610.9 microns / second (more than twice as fast as before infection).

[0305] Thus, in Case 2, the capillaries completely recovered from the effects of the novel coronavirus infection after drinking Leguan tea.

[0306] [Investigation]

[0307] In the capillaries of patients infected with the new coronavirus, capillary function is reduced due to microthrombosis (Lize M.et.al.Biosci Rep.2021Aug 27;41:8). In the infected persons of Case 1 and Case 2 in this embodiment, after being infected with the new coronavirus, the capillaries in the fingertips and nail folds were significantly atrophied, shortened, and thickened, and the blood flow rate was significantly slowed down, showing capillary damage caused by infection. By continuously drinking Leguan tea, the capillaries of both cases were completely restored, especially the length and blood flow rate of the capillaries were significantly improved, even exceeding the pre-infection level. This result shows that Leguan tea has an improvement effect on capillary damage in patients infected with the new coronavirus.

[0308] <Fifth Embodiment>

[0309] This example was carried out to verify the reproducibility of the Tie2 activation effect of the leaf extract of Cinnamon Bark (also known as Okinawa Karaki, hereinafter also referred to as "Okinawa Karaki").

[0310] [Purpose]

[0311] According to previous studies (Example 3), the Okinawa Karaki dried leaf extract extracted with methanol showed an effect of phosphorylating (activating) Tie2 at a concentration of 1000 ug / mL (the highest verified concentration of this test). Therefore, in this example, the effect of the Okinawa Karaki dried leaf extract in causing Tie2 phosphorylation at a concentration of 1000 ug / mL was reproducibly verified, and the effects of high concentrations of 1500 ug / mL and 2000 ug / mL of dried leaves and the effect of semi-dried leaf extract were also examined.

[0312] [Materials and Methods]

[0313] [Material]

[0314] Okinawa Karaki dried leaves and semi-dried leaf powder collected from the Ogimi farm in Okinawa Prefecture.

[0315] [method]

[0316] [1] Extraction method

[0317] Methanol extraction was performed by adding 60 L of methanol to 3 g of leaf powder, placing the mixture at 50°C for 3 hours, filtering the mixture, and concentrating the filtrate under reduced pressure.

[0318] [2] Analytical methods

[0319] Tie2 phosphorylation analysis was performed using cells overexpressing human Tie2 in mouse pro-B cells (Ba / F3) (Ba / F3-human Tie2). Ba / F3-human Tie2 was stimulated with Angiopoietin-1 (Ang1 300ng / ml), PBS or DMSO as negative control. Samples were directly added to culture medium containing 10% FBS RPMI1640 and 1pg / ml mouse IL-3 at different concentrations. After 15 minutes, cells were washed with PBS and cell extracts were collected using RIPA lysis buffer and Halt protease and phosphatase inhibitors. After 7.5% SDS gel electrophoresis, the extracts were transferred to nylon membranes, blocked in 4% skim milk powder + 0.5% BSA / TBST for 60 minutes, treated with HRP-labeled secondary antibodies, and developed with ECL colorimetric solution. Tie2 phosphorylation was detected using AmershamImager680 software.

[0320] [Test results]

[0321] The test results of this example are as follows Fig.29 , Fig.30 And as shown in Table 7. Fig.29 Western Blot diagram showing the detection results in this embodiment, Fig.30 Display data chart, Table 7 lists Fig.29 Western Blot values.

[0322] Table 7

[0323] Tie2 PTie2 PTie2 / Tie2 ①DMSO (10ul / mL) 566714 38876 0.068598976 1 ②)Ang1(300ng / mL)+DMSO(10ul / mL) 422770 359367 0.850029567 12.39129 ③B2 (1000ug / ml) 632630 82113 0.129796247 1.892102 ④B2 (1000ug / ml) 659359 44769 0.067897761 0.989778 ⑤) B2 (1500ug / ml) 680117 78460 0.115362504 1.681694 ⑥B2 (2000ug / ml) 590454 95427 0.161616316 2.355958 ⑦B2-2 (1000ug / ml) 678345 72530 0.106921994 1.558653 ⑧B2-2 (1000ug / ml) 747115 38890 0.052053566 0.758810 ⑨B2-2 (1500ug / ml) 648601 46793 0.072144508 1.051685 ⑩B2-2 (2000ug / ml) 587427 66850 0.113801374 1.658937

[0324] Fig.29 The phosphorylated Tie2 protein band detected by western blotting in each sample is shown.

[0325] like Fig.29As shown, the intensity of the detected band staining is proportional to the amount of phosphorylated Tie2 protein. The denser the band, the more phosphorylated Tie2 protein. No. (1) DMSO negative control, (2) Ang1 positive control, (3) dried leaf extract 1000ug / mL, (4) dried leaf extract 1000ug / mL, (5) dried leaf extract 1500ug / mL, (6) dried leaf extract 2000ug / mL, (7) semi-dried leaf extract 1000ug / mL, (8) semi-dried leaf extract 1000ug / mL, (9) semi-dried leaf extract 1500ug / mL, (10) semi-dried leaf extract 2000ug / mL.

[0326] like Fig.29 As shown, the concentration of phosphorylated Tie2 (P-Tie2) after treatment with dry leaf extract 1000ug / mL (3) was higher than that of DMSO control (1).

[0327] As shown in Table 7, the Tie2 phosphorylation rate was 1.9 times that of the DMSO control, and the reproducibility was confirmed (1.8 times in the previous study, and a phosphorylation rate exceeding 1.5 times indicated Tie2 phosphorylation).

[0328] Similarly, at concentrations of 1500 μg / mL (5) and 2000 μg / mL (6), the dried leaf extract induced Tie2 phosphorylation in a dose-dependent manner, and the EC50 of the dried leaf extract was estimated to be approximately 1000 μg / mL. Tie2 phosphorylation was also observed in semi-dried leaf extracts at 1000 ug / mL (7) and 2000 μg / mL (10) (the Tie2 phosphorylation rates were 1.6 and 1.7 times that of the DMSO control, respectively), and Tie2 phosphorylation was further confirmed by band concentration.

[0329] Table 7 shows the values ​​calculated by immuno-western blotting, and when the Tie2 phosphorylation rate is 1.5 times higher than that of the DMSO control, it indicates that Tie2 is phosphorylated. Tie2 phosphorylation was observed in the dry leaf extract at (3) 1000 ug / mL, (5) 1500 μg / mL, (6) 2000 μg / mL and in the semi-dried leaf extract at (7) 1000 ug / mL and (10) 2000 ug / mL.

[0330] Fig.30 The figure is a graph of the Tie2 phosphorylation data of Table 7. Compared with the DMSO control, the dry leaf extracts (3) 1000 ug / mL, (5) 1500 μg / mL, (6) 2000 μg / mL and the semi-dried leaf extracts (7) 1000 ug / mL and (10) 2000 μg / mL all showed Tie2 phosphorylation.

[0331] [Investigation]

[0332] By comparing with the DMSO control, the phosphorylation of Tie2 activated by Ang1 verified the effectiveness of the experiment in this example. In this example, the effect of inducing Tie2 phosphorylation was observed at a concentration of 1000 ug / mL using methanol from the dried leaves of Karaki in Okinawa.

[0333] To further verify its reproducibility and high concentration effect, additional tests were conducted using the same method, and semi-dried leaves of Okinawa Karaki were tested in the additional tests. The results are as follows:

[0334] [1] Compared to the DMSO control, 1000ug / mL of dried leaf extract increased phosphorylated Tie2 by 1.9-fold, and reproducibility was confirmed.

[0335] [2] Dry leaf extract 1500μg / mL and 2000μg / mL induced Tie2 phosphorylation in a dose-dependent manner.

[0336] [3] The EC50 of the dried leaf extract was estimated to be 1000 μg / mL.

[0337] [4] Semi-dried leaf extract also induced Tie2 phosphorylation in a dose-dependent manner at 1000ug / mL and 2000μg / mL.

[0338] [5] The EC50 of semi-dried leaf extract was also estimated to be 1000 μg / mL.

[0339] Therefore, this example shows that the dried and semi-dried leaf extracts of Okinawa Karaki have the effect of inducing the phosphorylation of Tie2. Since the semi-dried leaf sample has too high water and oil content and a low concentration of active ingredients, the Tie2 phosphorylation effect is weaker than that of the dried leaf extract.

[0340] <Conclusion>

[0341] A number of specific embodiments have been described above, but the present invention is not limited to these embodiments and can be modified or improved as needed.

[0342] [Possibility of industrial application]

[0343] The invention activates Tie2 receptors through active ingredients, promotes blood circulation, and improves the absorption rate in the body, thereby achieving capillary improvement. The invention can be used as a Tie2 receptor activator, a vascular maturation agent or a vascular stabilizer, and its food and drink, as well as a capillary improver and a capillary improving food and drink have wide application value.

[0344] The above description is for illustrative purposes only and is not intended to be limiting. Any equivalent modifications or changes made thereto without departing from the spirit and scope of the present invention shall be included in the present invention.

Claims

1. A Tie2 receptor activator containing an extract of Okinawa cinnamon leaves as an active ingredient.

2. A vascular maturation agent or a vascular stabilizer containing an extract of Okinawa cinnamon leaves as an effective ingredient, which has a vascular maturation or vascular stabilization effect based on Tie2 receptor activation.

3. A food or beverage for Tie2 receptor activation, vascular maturation and / or vascular stabilization, comprising the Tie2 activator according to claim 1, the vascular maturation agent or vascular stabilizer according to claim 2.

4. A capillary improving agent containing as active ingredients processed products of rooibos leaves and / or branches, processed products of Okinawa cinnamon leaves, processed products of branches and / or barks of Ceylon cinnamon and South China cinnamon, processed products of safflower, and processed products of black beans.

5. The capillary improving agent according to claim 4, wherein the processed product is a dried crushed product or a powder. The capillary improving agent according to claim 4 , further comprising lactic acid bacteria.

7. A food and drink for improving capillaries, characterized in that: Contains the capillary improving agent according to claim 4.

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