Potentilla anserine active composition for improving hypoxia insulin resistance and application thereof
Through the combination of fern butanol extract, fern ethanol extract and fern water-soluble extract, the insulin signaling pathway and oxidative stress response are regulated, the problem of insulin resistance in hypoxia environment is solved, and insulin resistance is improved and Effects of sugar metabolism disorders.
Patent Information
- Application Number
- CN202411610278.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-12
- Publication Date
- 2025-02-11
AI Technical Summary
In a hypoxic environment, the body will experience an insulin resistance reaction, resulting in a decrease in cell uptake and utilization of glucose, which will lead to glucose metabolism disorders and cell damage.
The combination of ferns, ferns, ferns, ferns, ferns, and ferns, water-soluble extract is used to regulate the insulin signaling pathway, enhance the activity of the AKT pathway, improve the uptake of glucose, and eliminate free radicals through antioxidants to regulate oxidative response Inspiring reaction.
Improve insulin resistance, enhance glucose uptake ability, maintain blood sugar balance, reduce sugar metabolism disorders, protect cells, and regulate oxidative stress response under hypoxia stress.
Smart Images
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of pharmaceutical compositions, and in particular relates to a potentilla active composition for improving hypoxia insulin resistance and an application thereof. Background Art
[0002] The plateau area is characterized by high altitude, low oxygen and cold climate. In this special low oxygen environment, the body will be subjected to low oxygen stress and the sugar metabolism function in the body will be disordered. Within a short period of time after entering the plateau area, the body's sympathetic nervous system will be excited and hormone secretion will increase, leading to a rapid increase in blood sugar stress. Moreover, as the time in the low oxygen environment increases, the peripheral tissue metabolism of glucose increases, liver gluconeogenesis is inhibited, and the fasting blood sugar level is significantly lower than that in the plains.
[0003] The main function of insulin is to lower blood sugar. It can only provide energy to the human body after blood sugar enters cells. Under the influence of hypoxia stress, insulin resistance will be induced, which will reduce the body's cells' uptake and utilization of glucose. Even if the insulin level in the blood increases, cells cannot effectively use glucose, and the body cannot maintain blood sugar at normal levels, thus causing cell damage and insufficient energy supply. Insulin resistance will also promote inflammatory response, further damage the insulin signaling pathway, and aggravate insulin resistance. Summary of the invention
[0004] The purpose of the present invention is to provide a potentilla active composition for improving hypoxia insulin resistance and its application, so as to solve the problem of insulin resistance reaction caused by hypoxia stress.
[0005] The purpose of the present invention can be achieved by the following technical solutions:
[0006] In a first aspect, the present invention provides a Potentilla anserine active composition for improving hypoxia insulin resistance, the Potentilla anserine active composition comprising 25 to 35 parts of Potentilla anserine n-butanol extract, 10 to 20 parts of Potentilla anserine ethanol extract, 15 to 25 parts of Potentilla anserine water-soluble extract and 5 to 10 parts of a composite regulating extract.
[0007] Under conditions of hypoxic stress, in order to adapt to environmental changes, the body will stimulate the activation of glycogen phosphorylase, causing a large amount of liver glycogen to break down and increase blood sugar. However, in a hypoxic state, insulin sensitivity is reduced, and the uptake and utilization of glucose is reduced, resulting in insulin resistance. It will also affect the insulin signaling pathway, further weakening the body's ability to take in glucose. Damage to the signaling pathway will further increase the degree of insulin resistance, leading to gradual glucose metabolism disorders in the body and causing cell damage.
[0008] By adopting the above technical scheme, the compound of various polar extracts of Potentilla anserina in the Potentilla anserina active composition can regulate the insulin signaling pathway and thus improve insulin resistance. Specifically, under hypoxic stress, the body will increase its glucose transport capacity in order to obtain more energy. GLUT4 can mediate glucose transmembrane transport and is the main step in glucose utilization and storage. Therefore, the number of GLUT4 binding sites in cells will increase, but the insulin resistance caused by hypoxia will repeatedly stimulate GLUT4 expression. Changes in GLUT4 expression quantity and quality will cause glucose metabolism disorders. At the same time, Akt2 protein is an important signaling molecule that promotes the translocation of GLUT4 vesicles to the cell membrane. Under hypoxic environment, AKT phosphorylation will be seriously hindered. As the main upstream signaling molecule of GLUT4, its dysfunction will not only affect glucose metabolism but also further increase the degree of insulin resistance, resulting in a vicious circle. The present invention finds that the combination of the n-butanol extract of Potentilla anserina, the ethanol extract of Potentilla anserina and the water-soluble extract of Potentilla anserina can well regulate the activity of the insulin signaling pathway, that is, enhance the activity of the AKT pathway, increase the phosphorylation level of AKT, thereby triggering the translocation of GLUT4 vesicles to the cell membrane, fusing with the cell membrane, increasing the uptake of glucose, and maintaining the body's insulin secretion-glucose consumption balance.
[0009] And under the condition of hypoxia stress, the body will have an oxidative stress reaction, that is, a large amount of reactive oxygen will be produced, resulting in a disorder of the balance of the antioxidant defense mechanism. When insulin resistance occurs, the body's ability to absorb glucose decreases, resulting in an increase in glucose content, further increasing the generation of reactive oxygen, which will reduce the secretion of insulin and increase the degree of insulin resistance. The combination of the n-butanol extract of Potentilla anserina, the ethanol extract of Potentilla anserina and the water-soluble extract of Potentilla anserina in the present invention can well regulate the oxidative stress state. The flavonoid components contained in the n-butanol extract of Potentilla anserina and the ethanol extract of Potentilla anserina and the antioxidant components contained in the water-soluble extract of Potentilla anserina can well scavenge free radicals, regulate oxidative stress-related factors and signal pathways, increase the activity of the Nrf2 signal pathway, the main regulator of oxidative stress, enhance the antioxidant capacity of cells, improve the level of oxidative stress, and reduce the effect of oxidative stress on insulin resistance.
[0010] In summary, the active composition of Potentilla anserina provided by the present invention can improve insulin resistance by regulating insulin signal transduction and glucose transport, enhance glucose uptake, maintain blood sugar balance, reduce the disordered state of glucose metabolism caused by insulin resistance, and protect cells from damage. At the same time, it can also regulate the oxidative stress response under hypoxia stress, inhibit the generation of oxygen free radicals and capture free radicals, and improve or inhibit the damage caused to the body by the hypoxic environment.
[0011] Preferably, the raw materials of the composite regulating extract include astragalus, mulberry leaf, American ginseng and rhodiola rosea in a mass ratio of 1: (0.6-0.8): (0.2-0.25): (0.4-0.5).
[0012] Preferably, the composite regulating extract is prepared according to the following method:
[0013] After mixing Astragalus and Rhodiola rosea, the alcohol extract and the residue are obtained by ethanol reflux extraction and filtration; the residue is mixed with mulberry leaves and American ginseng, and the water extract is obtained by water extraction and filtration; the water extract and the alcohol extract are mixed, concentrated, dried and ground to obtain a composite regulated extract.
[0014] By adopting the above technical scheme, the active composition of Potentilla anserine of the present invention is also added with a composite regulating extract, which is also extracted from plants. When combined with the polar extracts of Potentilla anserine, the side effects on the body are small and can compensate for the regulatory effect of the Potentilla anserine extract on cells.
[0015] Among them, the glycoside compounds and polysaccharides in Astragalus can enhance the activity of insulin signal transmission channels in cells, inhibit oxidative stress reactions, and improve insulin resistance; the active ingredients in mulberry leaves can increase cellular glucose consumption and cell activity, inhibit the occurrence of inflammation, and regulate the insulin signaling pathway; the saponin compounds in American ginseng can cooperate with the active ingredients in Rhodiola rosea to inhibit the production and capture of free radicals, inhibit liver gluconeogenesis, lower blood sugar, and improve insulin sensitivity.
[0016] The composite extract obtained by alcohol extraction and water extraction can synergize the active ingredients among each component, significantly enhance the overall antioxidant performance, lower blood sugar in the body, and improve insulin resistance. When combined with the polar active extracts of Potentilla anserina, it can enhance the regulation of the insulin pathway and reduce the impact of oxidative stress on the body, thereby improving the insulin resistance response under hypoxic stress conditions.
[0017] Preferably, the n-butanol extract of Potentilla anserina, the ethanol extract of Potentilla anserina and the water-soluble extract of Potentilla anserina are prepared according to the following method:
[0018] S1. The dried bracken was crushed and sieved to obtain bracken powder;
[0019] S2. Extracting the Pteridium anserine powder by reflux extraction with ethanol aqueous solution and concentrating under reduced pressure to obtain a pre-extract;
[0020] S3. The pre-extract was dispersed in deionized water, the temperature was raised to 50-60°C, and the mixture was stirred for 30-60 min, and then extracted with petroleum ether, ethyl acetate and n-butanol, and the n-butanol extract was concentrated under reduced pressure, freeze-dried and ground to obtain a n-butanol extract of Potentilla anserina;
[0021] S4. The supernatant obtained after extraction is concentrated under reduced pressure, and an ethanol aqueous solution is added to continue reflux extraction. The alcohol extract and the liquid phase are concentrated under reduced pressure, freeze-dried and ground to obtain a Potentilla anserina ethanol extract and a Potentilla anserina water-soluble extract, respectively.
[0022] Preferably, the mass fraction of the ethanol aqueous solution is 60-70%.
[0023] Preferably, the material-liquid ratio during the reflux extraction process is 1 g: (5-8) mL.
[0024] Preferably, the volume ratio of petroleum ether, ethyl acetate and n-butanol to the pre-extract aqueous solution during the extraction process is (3-5):1.
[0025] By adopting the above technical scheme, the present invention uses alcohol to extract the active ingredients therein, and then uses a variety of extractants to extract in sequence. Since Potentilla anserina includes various types of compounds, they will show different solubility in extractants with different polarities. In addition to using n-butanol for extraction, the present invention also uses petroleum ether and ethyl acetate for extraction, so as to selectively extract the active ingredients that are beneficial to improving insulin signal transduction, use other extractants to remove unnecessary impurities, improve the purity and activity of the Potentilla anserina n-butanol extract, and thus improve the ability of the Potentilla anserina active composition to improve insulin resistance.
[0026] And the extracted ingredients are further subjected to ethanol extraction, which can better utilize some water-soluble active ingredients in bracken anise. Some antioxidants such as vitamin C are water-soluble. These effective free radical scavengers can help the bracken anise active composition to scavenge oxygen free radicals and reduce oxidative stress, thereby achieving the effect of improving insulin resistance.
[0027] Preferably, the active composition of Potentilla anserina is prepared according to the following method: weigh the Potentilla anserina n-butanol extract, Potentilla anserina ethanol extract, Potentilla anserina water-soluble extract and composite regulated extract in corresponding parts by mass, mix them evenly, and grind them to obtain the active composition of Potentilla anserina.
[0028] In a second aspect, the present invention provides an application of a potentilla active composition for improving insulin resistance, wherein the potentilla active composition is applied to treat decreased insulin sensitivity caused by hypoxia stress.
[0029] In a third aspect, the present invention provides an application of a Potentilla anserine active composition for improving insulin resistance. The Potentilla anserine active composition is applied to insulin resistance reaction caused by hypoxia stress and / or to prevent insulin resistance.
[0030] Beneficial effects of the present invention:
[0031] 1. The combination of the bracken anserine n-butanol extract, the bracken anserine ethanol extract and the bracken anserine water-soluble extract contained in the bracken anserine active composition of the present invention can improve insulin resistance by regulating insulin signal transduction and glucose transport, enhance the ability to absorb glucose, regulate the oxidative stress response under hypoxic stress, and reduce the occurrence of glucose metabolism disorders, thereby improving or inhibiting the damage to the body caused by insulin resistance under hypoxic environment.
[0032] 2. The active composition of Potentilla anserina of the present invention also contains a composite regulating extract, which can compensate for the regulatory effect of various polar extracts of Potentilla anserina on the cellular insulin pathway, significantly enhance the overall antioxidant performance, reduce the impact of oxidative stress on the body, and thus improve the insulin resistance response under hypoxic stress conditions. DETAILED DESCRIPTION
[0033] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0034] Preparation Example
[0035] Preparation Example 1, a composite regulating extract is prepared according to the following method:
[0036] Take 100g of Astragalus and 45g of Rhodiola rosea and mix them, add 70% ethanol aqueous solution 10 times the amount of the medicinal materials, reflux and extract twice, the extraction time is 2h, and then filter to obtain the alcohol extract and the medicinal residue;
[0037] The residue was mixed with 70g of mulberry leaves and 25g of American ginseng, and extracted three times with 10 times the amount of the medicinal materials in water for 3 hours, and filtered to obtain a water extract;
[0038] The obtained water extract and alcohol extract are mixed evenly, concentrated, dried at 50°C, and finally ground to obtain a composite adjusted extract.
[0039] Preparation Example 2, a composite regulating extract, is different from Preparation Example 1 only in that the added amount of Rhodiola rosea is 40g, the added amount of mulberry leaf is 80g, and the added amount of American ginseng is 20g.
[0040] Preparation Example 3, a composite regulating extract, is different from Preparation Example 1 only in that the added amount of Rhodiola rosea is 50g, the added amount of mulberry leaf is 60g, and the added amount of American ginseng is 25g.
[0041] Preparation Example 4, a composite regulating extract, differs from Preparation Example 1 only in that Rhodiola rosea is not added.
[0042] Preparation Example 5, a composite regulated extract, differs from Preparation Example 1 only in that mulberry leaves are not added.
[0043] Preparation Example 6, a composite regulating extract, differs from Preparation Example 1 only in that American ginseng is not added.
[0044] Preparation Example 7, a composite regulating extract, differs from Preparation Example 1 only in that Rhodiola rosea and American ginseng are not added.
[0045] Preparation Example 8: An Astragalus extract was prepared according to the following method:
[0046] Take 100g of Astragalus and add 70% ethanol aqueous solution with 10 times the mass of Astragalus to reflux and extract twice for 2 hours, then filter to obtain alcohol extract, concentrate and dry the alcohol extract, and finally grind to obtain Astragalus extract.
[0047] Preparation Example 9, a Rhodiola rosea extract, prepared according to the following method:
[0048] Take 100g of Rhodiola rosea and add 70% ethanol aqueous solution with 10 times the mass of Rhodiola rosea for reflux extraction twice. The extraction time is 2h, and then the alcohol extract is obtained by filtration. The alcohol extract is concentrated and dried, and finally the Rhodiola rosea extract is obtained by grinding.
[0049] Preparation Example 10: A mulberry leaf extract was prepared according to the following method:
[0050] Take 100g of mulberry leaves, add 70% ethanol aqueous solution with 10 times the weight of mulberry leaves, reflux and extract twice, the extraction time is 2h, then filter to obtain alcohol extract, concentrate the alcohol extract, dry it, and finally grind it to obtain mulberry leaf extract.
[0051] Preparation Example 11, a Rhodiola rosea extract, prepared according to the following method:
[0052] Take 100g of American ginseng and add 70% ethanol aqueous solution with 10 times the mass of American ginseng for reflux extraction twice, the extraction time is 2h, then filter to obtain alcohol extract, concentrate the alcohol extract, dry it, and finally grind it to obtain American ginseng extract.
[0053] Example
[0054] Example 1, a Potentilla anserine active composition for improving insulin resistance, prepared according to the following method:
[0055] The dried bracken anserine was crushed and passed through a 100-mesh sieve to obtain bracken anserine powder;
[0056] Add 70% ethanol aqueous solution to the Pteridium anserine powder, with a solid-liquid ratio of 1 g:7 mL, perform reflux extraction for 3 times, the extraction time is 2 h, and then concentrate under reduced pressure to obtain a pre-extract;
[0057] The pre-extract was dispersed in 4 times the mass of deionized water, the temperature was raised to 60°C, and after stirring and mixing for 30 minutes, petroleum ether, ethyl acetate and n-butanol were used for extraction in sequence, wherein the volume ratio of petroleum ether, ethyl acetate and n-butanol to the aqueous solution of the pre-extract was 4:1, and the n-butanol extract was retained and concentrated under reduced pressure, freeze-dried and ground to obtain the n-butanol extract of Potentilla anserina.
[0058] The supernatant obtained after extraction was concentrated under reduced pressure, and an ethanol aqueous solution with a mass fraction of 70% was added, and reflux extraction was continued, wherein the solid-liquid ratio was 1g:7mL, reflux extraction was repeated twice, and the extraction time was 3h. The obtained alcohol extract and liquid phase were concentrated under reduced pressure, freeze-dried and ground to obtain the ethanol extract of Potentilla anserina and the water-soluble extract of Potentilla anserina, respectively.
[0059] The above-obtained Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and the composite regulating extract prepared in Preparation Example 1 were weighed in a mass ratio of 30:15:20:8, mixed evenly, and finally ground to obtain a Potentilla anserine active composition for improving insulin resistance.
[0060] Example 2, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass ratio of Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and the composite regulating extract prepared in Preparation Example 1 is 25:20:20:10.
[0061] Example 3, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass ratio of Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and the composite regulating extract prepared in Preparation Example 1 is 35:15:15:8.
[0062] Example 4, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass ratio of Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and the composite regulating extract prepared in Preparation Example 1 is 30:10:25:5.
[0063] Example 5, a Potentilla anserine active composition for improving insulin resistance, prepared according to the following method:
[0064] The dried bracken anserine was crushed and passed through a 100-mesh sieve to obtain bracken anserine powder;
[0065] Add 60% ethanol aqueous solution to the Pteridium anserine powder, with a solid-liquid ratio of 1 g:8 mL, perform reflux extraction for 3 times, the extraction time is 2 h, and then obtain a pre-extract by vacuum concentration;
[0066] The pre-extract was dispersed in 3 times the mass of deionized water, the temperature was raised to 60°C, and after stirring and mixing for 60 minutes, petroleum ether, ethyl acetate and n-butanol were used for extraction in sequence, wherein the volume ratio of petroleum ether, ethyl acetate and n-butanol to the aqueous solution of the pre-extract was 3:1, and the n-butanol extract was retained and concentrated under reduced pressure, freeze-dried and ground to obtain the n-butanol extract of Potentilla anserina;
[0067] The supernatant obtained after extraction was concentrated under reduced pressure, and an ethanol aqueous solution with a mass fraction of 70% was added, and reflux extraction was continued, wherein the solid-liquid ratio was 1g:7mL, reflux extraction was repeated twice, and the extraction time was 3h. The obtained alcohol extract and liquid phase were concentrated under reduced pressure, freeze-dried and ground to obtain the ethanol extract of Potentilla anserina and the water-soluble extract of Potentilla anserina, respectively.
[0068] The above-obtained Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and the composite regulating extract prepared in Preparation Example 2 were weighed in a mass ratio of 30:15:20:8, mixed evenly, and finally ground to obtain a Potentilla anserine active composition for improving insulin resistance.
[0069] Example 6, a Potentilla anserine active composition for improving insulin resistance, prepared according to the following method:
[0070] The dried bracken anserine was crushed and passed through a 100-mesh sieve to obtain bracken anserine powder;
[0071] Add 70% ethanol aqueous solution to the Pteridium anserine powder, with a solid-liquid ratio of 1 g:5 mL, perform reflux extraction for 3 times, the extraction time is 2 h, and then concentrate under reduced pressure to obtain a pre-extract;
[0072] The pre-extract was dispersed in 5 times the mass of deionized water, the temperature was raised to 60°C, and after stirring and mixing for 30 minutes, petroleum ether, ethyl acetate and n-butanol were used for extraction in sequence, wherein the volume ratio of petroleum ether, ethyl acetate and n-butanol to the aqueous solution of the pre-extract was 5:1, and the n-butanol extract was retained and concentrated under reduced pressure, freeze-dried and ground to obtain the n-butanol extract of Potentilla anserina.
[0073] The supernatant obtained after extraction was concentrated under reduced pressure, and an ethanol aqueous solution with a mass fraction of 70% was added, and reflux extraction was continued, wherein the solid-liquid ratio was 1g:7mL, reflux extraction was repeated twice, and the extraction time was 3h. The obtained alcohol extract and liquid phase were concentrated under reduced pressure, freeze-dried and ground to obtain the ethanol extract of Potentilla anserina and the water-soluble extract of Potentilla anserina, respectively.
[0074] The above-obtained Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and the composite regulating extract prepared in Preparation Example 3 were weighed in a mass ratio of 30:15:20:8, mixed evenly, and finally ground to obtain a Potentilla anserine active composition for improving insulin resistance.
[0075] Example 7, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the compound regulating extract prepared in Preparation Example 1 is replaced by an equal amount of the compound regulating extract prepared in Preparation Example 4.
[0076] Example 8, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the compound regulating extract prepared in Preparation Example 1 is replaced by an equal amount of the compound regulating extract prepared in Preparation Example 5.
[0077] Example 9, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the compound regulating extract prepared in Preparation Example 1 is replaced with an equal amount of the compound regulating extract prepared in Preparation Example 6.
[0078] Example 10, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the compound regulating extract prepared in Preparation Example 1 is replaced with an equal amount of the compound regulating extract prepared in Preparation Example 7.
[0079] Example 11, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that, in the preparation process of Potentilla anserine n-butanol extraction, only n-butanol is used for extraction, and the volume ratio of n-butanol to the pre-extract aqueous solution is 4:1. The obtained n-butanol extract is concentrated under reduced pressure, freeze-dried and ground to obtain Potentilla anserine n-butanol extract.
[0080] Comparative Example
[0081] Comparative Example 1, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of the Potentilla anserine n-butanol extract is 20 parts.
[0082] Comparative Example 2, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of the Potentilla anserine n-butanol extract is 40 parts.
[0083] Comparative Example 3, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of Potentilla anserine ethanol extract is 5 parts.
[0084] Comparative Example 4, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of Potentilla anserine ethanol extract is 25 parts.
[0085] Comparative Example 5, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of the Potentilla anserine water-soluble extract is 10 parts.
[0086] Comparative Example 6, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of the Potentilla anserine water-soluble extract is 30 parts.
[0087] Comparative Example 7, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of the composite regulating extract prepared in Preparation Example 1 is 2 parts.
[0088] Comparative Example 8, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the mass fraction of the composite regulating extract prepared in Preparation Example 1 is 15 parts.
[0089] Comparative Example 9, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the Potentilla anserine n-butanol extract is not added to the Potentilla anserine active composition.
[0090] Comparative Example 10, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that no Potentilla anserine ethanol extract is added to the Potentilla anserine active composition.
[0091] Comparative Example 11, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that no Potentilla anserine water-soluble extract is added to the Potentilla anserine active composition.
[0092] Comparative Example 12, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that an equal amount of the Astragalus extract prepared in Preparation Example 8 is used to replace the composite regulating extract prepared in Preparation Example 1.
[0093] Comparative Example 13, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that an equal amount of the Rhodiola rosea extract prepared in Preparation Example 9 is used to replace the composite regulating extract prepared in Preparation Example 1.
[0094] Comparative Example 14, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that an equal amount of the mulberry leaf extract prepared in Preparation Example 10 is used to replace the composite regulating extract prepared in Preparation Example 1.
[0095] Comparative Example 15, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that an equal amount of the American ginseng extract prepared in Preparation Example 11 is used to replace the composite regulating extract prepared in Preparation Example 1.
[0096] Comparative Example 16, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the composite regulating extract prepared in Preparation Example 1 is not added to the Potentilla anserine active composition.
[0097] Comparative Example 17, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the Potentilla anserine active composition is only the Potentilla anserine n-butanol extract.
[0098] Comparative Example 18, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the Potentilla anserine active composition is only the Potentilla anserine ethanol extract.
[0099] Comparative Example 19, a Potentilla anserine active composition for improving insulin resistance, is different from Example 1 only in that the Potentilla anserine active composition is only a water-soluble extract of Potentilla anserine.
[0100] Performance testing
[0101] A total of 160 6-week-old healthy male Sprauge-Dawley (SD) rats with an average body weight of (190±9) g were selected and divided into a normal group (5 rats), a control group (5 rats) and an experimental group (30×5 rats).
[0102] The control group and the experimental group were placed in a hypoxic chamber with the oxygen concentration controlled at (16±2)% to simulate the plateau hypoxic environment.
[0103] The experimental group was given a dose of 0.2g / kg, and the control group and the normal group were given the same dose of normal saline. They were kept in a hypoxic environment for 6 hours, during which they had free access to water but were fasting; the normal group was raised in a normal environment and was fasting but not water deprivation.
[0104] Then, the blood glucose level of the rats and the expression of GLUT4 and AKT2 proteins in the skeletal muscle cytoplasm were tested respectively, and the insulin resistance index was simulated and evaluated (insulin resistance index = (fasting blood glucose × fasting insulin) / 22.5). The test results were averaged.
[0105] The above test results are shown in Table 1 and Table 2:
[0106] Table 1 Blood glucose and insulin resistance index test results
[0107]
[0108]
[0109] Table 2 Comparison of GLUT4 and AKT2 protein expression
[0110]
[0111] According to Table 1 and Table 2, in combination with Example 1, Example 2 to Example 6, it can be seen that the blood sugar level, insulin resistance degree, and GLUT4 and AKT2 protein expression of Example 2 to Example 6 are not significantly different from those of Example 1, indicating that there is no significant difference in the ability to improve insulin resistance of Example 2 to Example 6. The reason is that compared with Example 1, Example 2 to Example 6 only changed the raw materials, raw material ratios, and process parameters within the required range, which has little effect on the ability of the final Potentilla anserine active composition to improve insulin resistance.
[0112] Combining Example 1, Example 7 to Example 10, it can be seen that the blood sugar level, insulin resistance, and GLUT4 and AKT2 protein expression in Example 7 to Example 10 are increased compared to Example 1, and the change in Example 8 is more obvious. The reason is that Examples 7 to Example 10 change the composition of the composite regulating extract, and reduce one or two types of extracts, resulting in a decrease in the improvement effect, while in Example 8, mulberry leaves are not added, and the improvement of the insulin signaling pathway is too reduced, resulting in a more obvious increase in the expression of GLUT4 and AKT2 proteins.
[0113] Combining Example 1 and Example 11, it can be seen that the blood sugar level, insulin resistance, and GLUT4 and AKT2 protein expressions of Example 11 are increased compared with those of Example 1. The reason is that in the preparation process of the n-butanol extract of Potentilla anserina in Example 11, only n-butanol is used for extraction, resulting in an increase in the impurity content in the n-butanol extract, a decrease in activity, and a decrease in the effect of improving insulin resistance.
[0114] In combination with Example 1 and Comparative Examples 1 to 8, it can be seen that the blood sugar levels, insulin resistance, and GLUT4 and AKT2 protein expressions of Comparative Examples 1 to 8 are significantly increased compared to those of Example 1, among which the changes in Comparative Example 1 are more obvious. The reason is that Comparative Examples 1 to 8 adjust the addition amount of each component of the active composition of Potentilla anserina compared to Example 1, and the change in components will cause the expression of GLUT4 and AKT2 proteins to be affected, whether positive or negative, excessive stimulation or guidance will cause abnormal insulin signal transduction and cause insulin resistance. In Comparative Example 1, the content of the n-butanol extract of Potentilla anserina was reduced, and the ability of the active composition of Potentilla anserina to regulate insulin and oxidative stress reactions decreased significantly, resulting in a decrease in the ability to improve insulin resistance.
[0115] Combining Example 1 and Example 9 to Example 11, it can be seen that the blood sugar level, insulin resistance, and GLUT4 and AKT2 protein expressions of Examples 9 to 11 are significantly increased compared to Example 1. The reason is that the active compositions of Potentilla anserinae in Examples 9 to 11 have reduced one of the polar extracts of Potentilla anserinae, resulting in a decrease in the ability of the active composition of Potentilla anserinae to regulate glucose uptake balance, insulin signal transduction, and oxidative stress response, thereby resulting in a decrease in the ability of the active composition of Potentilla anserinae to improve insulin resistance.
[0116] Combining Example 1 and Example 12 to Example 16, it can be seen that the blood sugar level, insulin resistance, and GLUT4 and AKT2 protein expression of Examples 12 to Example 16 are significantly increased compared to Example 1, and the change in Example 16 is the most obvious. The reason is that Examples 12 to Example 15 use single-component extracts to replace the composite regulating extracts. Compared with the composite synergistic effect between the components, the single component has a reduced ability to improve the body's ability to improve insulin resistance and scavenge free radicals; Example 16 does not add a composite regulating extract, so it is difficult to coordinate and compensate for the regulatory effect of the polar extract of Potentilla anserina, and the scavenging effect on free radicals and the effect of improving insulin sensitivity are reduced.
[0117] Combining Example 1 and Example 17 to Example 19, it can be seen that the blood sugar level, insulin resistance, and GLUT4 and AKT2 protein expressions of Example 17 to Example 19 are significantly increased compared to Example 1. The reason is that compared to Example 1, Example 17 to Example 19 only added one of the polar extracts of Potentilla anserina, lacking the regulation of the insulin signaling channel or reducing the scavenging effect on free radicals, which will weaken the improvement effect of the composition on insulin resistance.
[0118] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.
[0119] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A potentilla anserine active composition for improving hypoxia insulin resistance, characterized in that: The active composition of Potentilla anserine comprises 25 to 35 parts of Potentilla anserine n-butanol extract, 10 to 20 parts of Potentilla anserine ethanol extract, 15 to 25 parts of Potentilla anserine water-soluble extract and 5 to 10 parts of compound regulating extract.
2. The active composition of Potentilla anserina for improving hypoxia insulin resistance according to claim 1, characterized in that: The raw materials of the composite regulating extract include astragalus, mulberry leaf, American ginseng and rhodiola rosea in a mass ratio of 1: (0.6-0.8): (0.2-0.25): (0.4-0.5).
3. The active composition of Potentilla anserina for improving hypoxia insulin resistance according to claim 2, characterized in that: The composite regulating extract is prepared according to the following method: After mixing Astragalus and Rhodiola rosea, the alcohol extract and the residue are obtained by ethanol reflux extraction and filtration; the residue is mixed with mulberry leaves and American ginseng, and the water extract is obtained by water extraction and filtration; the water extract and the alcohol extract are mixed, concentrated, dried and ground to obtain a composite regulated extract.
4. The active composition of Potentilla anserina for improving hypoxia insulin resistance according to claim 1, characterized in that: The bracken anserine n-butanol extract, bracken anserine ethanol extract and bracken anserine water-soluble extract are prepared according to the following method: S1. The dried bracken was crushed and sieved to obtain bracken powder; S2. Extracting the Pteridium anserine powder by reflux extraction with ethanol aqueous solution and concentrating under reduced pressure to obtain a pre-extract; S3. The pre-extract was dispersed in deionized water, the temperature was raised to 50-60°C, and the mixture was stirred for 30-60 min, and then extracted with petroleum ether, ethyl acetate and n-butanol, and the n-butanol extract was concentrated under reduced pressure, freeze-dried and ground to obtain a n-butanol extract of Potentilla anserina; S4. The supernatant obtained after extraction is concentrated under reduced pressure, and an ethanol aqueous solution is added to continue reflux extraction. The alcohol extract and the liquid phase are concentrated under reduced pressure, freeze-dried and ground to obtain a Potentilla anserina ethanol extract and a Potentilla anserina water-soluble extract, respectively.
5. The active composition of Potentilla anserina for improving insulin resistance according to claim 4, characterized in that: The mass fraction of the ethanol aqueous solution is 60-70%.
6. The active composition of Potentilla anserina for improving insulin resistance according to claim 4, characterized in that: The material-liquid ratio in the reflux extraction process is 1 g: (5-8) mL.
7. The active composition of Potentilla anserina for improving insulin resistance according to claim 4, characterized in that: In the extraction process, the volume ratio of petroleum ether, ethyl acetate and n-butanol to the pre-extract aqueous solution is (3-5):
1.
8. The active composition of Potentilla anserina for improving hypoxia insulin resistance according to claim 1, characterized in that: The active composition of Potentilla anserine is prepared according to the following method: the Potentilla anserine n-butanol extract, Potentilla anserine ethanol extract, Potentilla anserine water-soluble extract and composite regulated extract are weighed in corresponding parts by mass, mixed evenly, and ground to obtain the active composition of Potentilla anserine.
9. An application of the active composition of Potentilla anserinae for improving insulin resistance according to claim 1, characterized in that: The active composition of Potentilla anserina is used to reduce insulin sensitivity caused by hypoxia stress.
10. An application of the active composition of Potentilla anserinae for improving insulin resistance according to claim 1, characterized in that: The potentilla anserine active composition is used for treating insulin resistance reaction caused by hypoxia stress and / or preventing insulin resistance.