CN102652757B - Application of health-care drinking liquid containing deep seawater in preparation of medicaments or health-care products for preventing or treating metabolic syndrome - Google Patents
Application of health-care drinking liquid containing deep seawater in preparation of medicaments or health-care products for preventing or treating metabolic syndrome Download PDFInfo
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Abstract
The invention provides application of health-care drinking liquid containing deep seawater in preparation of medicaments or health-care products for preventing or treating metabolic syndrome. The health-care drinking liquid comprises liquid A and liquid B, wherein the liquid A is fresh water; and the liquid B is high-concentration concentrate obtained by reduced pressure concentration of concentrated water separated from the deep seawater through a reverse osmosis device, and the hardness range of the liquid B is 200 to 1000ppm. The health-care drinking liquid can regulate the insulin signaling channel in vitro level and improve the resistance of insulin so as to prevent the metabolic syndrome. The health-care drinking liquid prepared from the deep seawater has the advantages of rich resources, easiness in industrialization and the like, and has broad market and application prospect on the aspect of preventing and treating the metabolic syndrome.
Description
Technical field
The invention belongs to the marine biotechnology field, relate to the comprehensive utilization of deep seawater, be specifically related to a kind of healht-care liquid of deep seawater that contains in preparation prevention or the medicine for the treatment of metabolic syndrome or the application in the health product.
Background technology
Metabolic syndrome is one group and assembles morbidity with central obesity, hyperglycemia (diabetes or IGR), hyperlipidemia and hypertension etc., have a strong impact on the clinical syndrome of body health, merge with multiple metabolic disease and to appear as clinical characters, be one group in the be mutually related combination of risk factor of metabolism.In recent years, along with economy develops rapidly, life and diet style westernization, the prevalence of metabolic syndrome raises.Metabolic syndrome day by day becomes China, especially the important public health problem in big and medium-sized cities.Studies show that insulin resistant (IR) is the pathogenesis basis of metabolic syndrome, insulin resistant has become a research focus of medical domain in recent years.
Insulin resistant refers to that a certain amount of insulin is combined the artifact effect and is lower than normally with its specific receptor, be the effect that the insulin of physiological concentration does not reach expection, mainly show as insulin and suppress liver and discharge the ability of glucose and promote surrounding tissue (mainly being skeletal muscle and fat) to absorb and utilize the ability of glucose to descend.
Liver is the critical organ of insulin resistant.The liver insulin resistant refers to that mainly insulin suppresses the ability decline of liver glucose output.Glycogen output mainly is made up of glyconeogenesis and glycogenolysis two parts.Glyconeogenesis refers to synthesis of glucose such as nonsugar such as lactic acid, 1-propenol-3, glucogenic amino acid, glycerol; Glycogenolysis refers to employ the glycogen that is stored in the liver glucose is provided.Liver is kept generation and output and sugared absorption, utilization and the storage in feed back of the endogenous sugar under the empty stomach state as the main target organ of insulin action.In recent years liver insulin resistant Study on Mechanism has been become focus.
The IRS-2/PI3K signal is that insulin is at the main signal transduction pathway of liver performance physiological effect.After the feed of animal or human's class, go into the Insulin receptor INSR (InsR) that blood acts on the liver plasma membrane surface by the insulin that the beta Cell of islet secretion produces, the tyrosine site autophosphorylation that is located at intracytoplasmic β subunit activates.After making IRS (IRS-2) tyrosine site phosphorylation, the Insulin receptor INSR that activates activates, and then activation phosphatidylinositols 3 kinases (PI3K), PI3K after the activation can catalysis 4,5-diphosphonic acid phosphatidylinositols (PIP2) generates triphosphoric acid phosphatidylinositols (PIP3), it is as second message,second messenger's activated protein kinase B (Akt), and the Akt of activation participates in the metabolism regulating action of performance insulin by following approach: (1) plays an important role by the transposition to glucose transporter GLUT4 vesicle in the cell.Under insulin stimulating, GLUT4 vesicle pond in the cell moves to cell membrane, merges with film then, the GLUT4 molecule is fixed on the cell membrane, thus the effect of performance transhipment glucose; (2) promote glycogen synthetic by glycogen synthase kinase-3 (GSK-3).GSK-3 is the physiology substrate of first found PKB/Akt, and PI3K, PDK, Akt and GSK3 have formed insulin and regulated an important branch road in the synthetic signal cascade system of glycogen.The isomer GSK3 α of the GSK3 of two kinds of forms and the aminoterminal of GSK3 β all contain the phosphorylation site of Akt, and insulin promotes glycogen synthetic by phosphorylation and inactivation GSK3; (3) expression and the glyconeogenesis of inhibition glyconeogenesis gene G-6-Pase (G6Pase) and PCK (PEPCK), final blood sugar lowering.
Insulin receptor INSR signal transduction defective is the main cause that IR takes place, and the key of normal insulin signaling conduction is the tyrosine phosphorylation process on the various kinase proteins.In most insulin resistants, the tyrosine phosphorylation of Insulin receptor INSR is normal substantially, the key position that insulin resistant takes place is IRS, IRS-1, the IRS-2 main signal protein after as InsR wherein, the reduction of its gene expression, the minimizing of protein expression and phosphorylation obstacle all can cause its activation to PI3K obviously to descend, and have hindered the conduction of insulin signaling pathway downstream signal thus and cause IR.
The energy metabolism dysequilibrium is the major reason that IR takes place.Protein kinase (AMPK) signal path that adenosine phosphate (AMP) activates is the key link of regulating the cellular energy state, change that can perception cellular energy metabolism state, and keep cellular energy demand balance by a plurality of links that influence the cellular material metabolism.Activate after the AMPK phosphorylation, active A MPK activates the multiple target molecule in its downstream, closes metabolic pathway of synthesizing thereby reach, and opens the effect of catabolic pathway.At skeletal muscle, glucose uptake and fatty acid oxidation are increased, and promote mitochondrial biosynthesis; At liver, suppress the synthetic of glucose and lipid, promote lipid oxidation; In fatty tissue, reduced lipid decomposition and lipid and generated.Otherwise when AMPK dephosphorylation inactivation, the body anabolism is occupied an leading position, lipid within endothelial cells, the synthetic increase of glycogen.Under the normal physiological state, the AMPK of activation regulates and control its downstream signal pathway, increase on the one hand the transposition of GLUT4 and enhances skeletal flesh to the reaction of insulin, motion and metformin improve the type 2 diabetes mellitus patient by this approach exactly; On the other hand, active A MPK can suppress Hydroxymethylglutaryl list acyl coenzyme A reductase (HMGCR), acetyl-CoA carboxylase lipid synthase activities such as (ACC), reduce the sterin regulating element in conjunction with the expression of albumen (SREBP)-lc and fatty acid synthase (FAS), thereby inhibition lipogenesis, increase the fatty acid oxidation approach simultaneously, reduce liver inner lipid content.In addition, behind the AMPK pathway activation, the downstream can be by strengthening mitochondrial function, promoting mitochondrion propagation to promote the energy utilization.Therefore, the damage of AMPK approach can cause body IR and lipid dystopy deposition.
Oxidative stress is that insulin resistant takes place and a major reason that develops.Long-term hyperglycemia, hyperlipidemia can cause mitochondrion to produce a large amount of reactive oxygen specieses (ROS), the generation of a large amount of ROS and accumulate in tissue, remove as the antioxidant system in can not being organized, will pass through the activation of IKK, NF-κ B, systems such as p38MAKP, JNK/SAPK, the signal path in interference cell Insulin receptor INSR signal transduction and downstream such as Akt, Insulin receptor INSR and receptor substrate phosphorylation finally cause insulin resistant.
Mineral element plays a significant role in the generation of IR, evolution.Found through experiments magnesium deficiency as Su á res etc. reduces relevant with the activity of insulin receptor tyrosine kinase; Chromium can be expressed and influence content of insulin, increases Insulin receptor INSR quantity, improves beta Cell of islet sensitivity, strengthens and take the photograph and activate insulin receptor tyrosine kinase in the adhesion, enhancing insulin of insulin and increase insulin sensitivity by regulating insulin gene; Zinc, selenium then can by the oxidation resistance that improves body accelerate the removing of free radical and inhibited oxidation stress, thereby improve insulin resistant.
Deep seawater typically refer to solar radiation less than, can not carry out photosynthesis, the sea water that about 200m degree of depth is following.Deep seawater is in the ocean that no sunlight enters does not have photosphere, and away from influence and pollution from the chemical substance of human, land and atmosphere.Studies show that deep seawater has following main feature: (1) low temperature homeostasis-be not subjected to solar radiation, changeable unlike the sea surface coolant-temperature gage, deep seawater is temperature-resistant all the year round, is constant at about 5 ℃; (2) the abundant and stability of mineral element-with sea surface water ratio, deep seawater is rich in 92 kinds of mineral elements such as the necessary calcium of human body, magnesium, potassium, sodium, ferrum, copper, lithium, molybdenum, manganese, zinc, germanium, iodine, phosphorus, fluorine, selenium, trivalent chromium, silicate ion, has contained the required element kind of human body metabolism.Except its content is abundant, because these water flow in the bathypelagic of " no photosphere " with the very long time, ectocine with taking place, not being subjected in unglazed cooperation, and therefore the composition of contained mineral element is very stable, has ideally solved the requirement that total balance of the body is absorbed various required mineral elements; (3) aseptic spatter property-the be in deep water of ocean " no photosphere " except away from the influence of human modern civilization and be not subjected to the pollution of land, Atmospheric Chemistry material, pathogenic bacteria, itself does not have the condition that generates antibacterials such as pathogen yet.Therefore, deep seawater is the water of the aseptic nature that cleans very much; (4) deep seawater water quality is alkalescence.
Owing to have above characteristics, deep seawater is being subjected to attracting attention of common people as a kind of new natural resources.Deep seawater is rich in various mineral and trace element, and it has good improvement effect to insulin resistant and metabolic syndrome in theory.China's marine site broadness, the coastline is very long, and marine resources are abundant, but still do not have the research of deep seawater Application and Development aspect; And China's water scarcity, water is seriously polluted, and the bathypelagic water resource of exploitation China has very important meaning.
China's marine site broadness, the coastline is very long, and marine resources are abundant, but still do not have the research of deep seawater Application and Development aspect; And China's water scarcity, water is seriously polluted, and the bathypelagic water resource of exploitation China has very important meaning.
Summary of the invention
, water in short supply day by day at present China water resource pollutes and is on the rise and the rapid hot issue such as risings of metabolic syndrome sickness rate, the invention provides a kind of healht-care liquid that contains deep seawater in the preparation prevention or treats the medicine of metabolic syndrome or the application in the health product.
For achieving the above object, the present invention adopts following technical proposals to be achieved:
A kind of healht-care liquid of deep seawater that contains is in preparation prevention or the medicine for the treatment of metabolic syndrome or the application in the health product, described healht-care liquid comprises A liquid and B liquid, described A liquid is fresh water, described B liquid makes through following steps: deep seawater is isolated dense water section behind reverse osmosis unit, dense water section is carried out concentrating under reduced pressure, make its volume be concentrated into the 0.67-3.33% of original volume, remove by filter the sodium chloride of separating out then, the height concentrated solution that obtains is B liquid, the hardness of described healht-care liquid is at 200-1000ppm, and described deep seawater is the following and sea water after desalting processing of sea level 500m.
To further improvement in the technical proposal: magnesium in the described healht-care liquid: calcium: potassium: the concentration ratio of sodium element is 2.5-3.5: 0.45-1: 0.3-1: 0.3-1.5, do not contain Organic substance, and water quality is alkalescence.
To further improvement in the technical proposal: described magnesium: calcium: the ratio of greater inequality of the concentration of potassium is 3: 1: 1.
To further improvement in the technical proposal: the water quality of described healht-care liquid is alkalescence, the pH value 7.5 ± 0.3 of 25 ℃ of mensuration.
To further improvement in the technical proposal: the beneficial element content in the described healht-care liquid is: zinc 0.0325-16.4 μ g/L; Selenium 0.0395-0.712 μ g/L; Chromium 0.104-4.54 μ g/L; Manganese 0.156-5.52 μ g/L.
To further improvement in the technical proposal: the harmful element content in the described healht-care liquid is: lead＜5 μ g/L; Hydrargyrum＜0.5 μ g/L; Arsenic＜5 μ g/L; Cadmium＜1 μ g/L.
To further improvement in the technical proposal: described deep seawater is taken from apart from beyond the 20km of coastline.
To further improvement in the technical proposal: described fresh water sources Yu Haiyang deep water is isolated fresh water after the reverse osmosis unit circulation.
To further improvement in the technical proposal: it is the healht-care liquid of 600-1000ppm that diet is selected hardness for use to the less metabolic syndrome patient of the picked-up of mineral, is the healht-care liquid of 200-400ppm otherwise select hardness.
Compared with prior art, advantage of the present invention and good effect are:
The healht-care liquid that contains deep seawater provided by the invention is rich in healthy trace elements with household magnesium, calcium, vanadium, chromium, manganese, zinc, selenium, and the content of poisonous trace element hydrargyrum and lead is very low, the present invention shows by biotic experiment, this healht-care liquid does not have toxicity at cellular level, safe, and can promote the synthetic of glycogen, thereby reduce glycogen output; Can improve insulin resistant by the lipid route of synthesis that activates in the AMPK inhibition liver; Can also improve insulin resistant by the oxidation resistance of enhancing body.Significantly regulate insulin signaling pathway by these approach, metabolic diseases such as metabolic syndrome are had preventive and therapeutic action preferably.
The specific embodiment
Below in conjunction with the specific embodiment technical scheme of the present invention is described in further detail.
Embodiment 1
One, the preparation of healht-care liquid
The preparation method of healht-care liquid of the present invention is as follows:
1) bathypelagic water pretreatment: will take from apart from beyond the 20km of coastline, sea level 500m puts into water tank with dark deep seawater, adopts filter to filter, and removes phytoplankton and microorganism in the sea water, and by the active carbon filter filtration, further remove Organic substance.
2) fresh water and dense separated form water: the deep seawater that obtains after the pretreatment is entered reverse osmosis unit by high-pressure pump deep seawater is divided into the fresh water part (fresh water part water quality is identical with pure water, or water quality is near pure water with dense water section; The concentration of various zwitterions is near 2 times of various ion concentrations in the former sea water in the dense water section), isolated fresh water partly is that A liquid enters the fresh water water tank, isolated dense water section enters dense water water tank.The technological parameter of reverse osmosis unit is: 4MPa, power 1.5KW, aquifer yield 1T/d are pressed in running.
3) dense water takes off the sodium processing: the dense water section in the dense water water tank is carried out concentrating under reduced pressure, make its volume be concentrated into the 0.67-3.33% of original volume, remove by filter the sodium chloride of separating out then, the concentrated solution that is highly concentrated is B liquid, magnesium in the described concentrated solution: calcium: potassium: the concentration ratio of sodium element is 2.5-3.5: 0.45-1: 0.3-1: 0.3-1.5, and described magnesium, calcium, potassium, the content concn unit of sodium element in concentrated solution are mg/L.Trace element chromium wherein, zinc, selenium, the concentration of manganese etc. and cycles of concentration are the multiple proportions relation.
Perhaps, also can adopt the selectivity electrodialysis to reverse osmosis after isolated dense water section take off sodium and handle, electrodialysis plant adopts 1 valency cation selective to see through film, cavity block adopts the conventional ion exchange membrane.All aniones all can see through cavity block in electrodialytic process, and only univalent cation sees through anode membrane, and the polyvalent cation that takes off in the sodium dense water afterwards is close with the concentration in the former dense water.
4) healht-care liquid fill: the fresh water A liquid that will come out through the step 1) reverse osmosis unit and the resulting B liquid of step 3) can according to demand mixed the healht-care liquid of different hardness, described hardness is with CaCO
3Cubage (GB 5749-2006 drinking water sanitary standard), hardness=Ca (ppm)) * 2.5+Mg (ppm) * 4.1 its computing formula is:, through high temperature sterilize, packing gets final product.
Adopt inductively coupled plasma mass spectrum (ICP-MS) that the inorganic element content in the healht-care liquid is measured: be that 1000 healht-care liquid is example with hardness, wherein several main mine materials and micronutrient levels are: magnesium: 200-224mg/L; Calcium: 33-67mg/L; Potassium: 20-67mg/L; Sodium: 20-100mg/L (content of sodium is more low more good); Zinc: 0.0325-16.4 μ g/L; Selenium: 0.0395-0.712 μ g/L; Chromium: 0.104-4.54 μ g/L; Manganese 0.156-5.52 μ g/L, lead＜5 μ g/L; Hydrargyrum＜0.5 μ g/L; Arsenic＜5 μ g/L; Cadmium＜1 μ g/L.
Healht-care liquid mineral under the 200-1000 hardness of the present invention and micronutrient levels are directly proportional with its hardness.Magnesium in the described healht-care liquid: calcium: potassium: the concentration ratio of sodium element is 2.5-3.5: 0.45-1: 0.3-1: 0.3-1.5, is rich in healthy trace elements with household zinc, selenium, chromium, manganese, poisonous trace element lead, hydrargyrum, arsenic, cadmium content is lower, do not contain Organic substance, and water quality is alkalescence.
Described healthy trace elements with household content is as follows: zinc: 0.0325-16.4 μ g/L; Selenium: 0.0395-0.712 μ g/L; Chromium: 0.104-4.54 μ g/L; Manganese 0.156-5.52 μ g/L.
Described poisonous micronutrient levels is as follows: lead＜5 μ g/L; Hydrargyrum＜0.5 μ g/L; Arsenic＜5 μ g/L; Cadmium＜1 μ g/L.
Use pH meter that its pH value is measured (25 ℃), the result shows healht-care liquid pH value in 7.5 ± 0.3 scopes, and water quality is alkalescence.
Two, healht-care liquid is to improvement effect and the mechanism of insulin signaling pathway and insulin resistant
1, experimental technique
1) cell culture: human liver cancer cell HepG2 cell, cell culture medium are low sugar DMEM (5.5mmol/L) culture medium that contains 10% hyclone, and culture environment is 37 ℃, 5%CO
2, went down to posterity once in 1: 3 ratio in per 3 days.
2) healht-care liquid is to the cytotoxicity of HepG2 cell: collect the logarithmic (log) phase cell, adjust concentration of cell suspension and be inoculated in 96 orifice plates, 10000 cells in every hole.37 ℃, 5%CO
2Cultivate, treat that cell degree of converging reaches at 70% o'clock, be changed to the DMEM that does not contain serum, hunger is spent the night, and adds the healht-care liquid of different hardness as shown in table 1 then, and each concentration is established 6 parallel holes.Continue to cultivate 24h, every hole adds freshly prepared 20 μ L MTT (5mg/mL), continues to cultivate 4h.Sop up supernatant then, every hole adds 150 μ L DMSO, low-speed oscillation 10min, and abundant dissolving crystallized thing is surveyed the OD value with enzyme-linked immunosorbent assay instrument at 570-630nm, and the record result calculates the cell inhibiting rate.
3) foundation of insulin resistant HepG2 (IR-HepG2) cell model and administration are handled: the HepG2 cell of the trophophase of taking the logarithm, be made into the individual cells suspension with the DMEM culture fluid that contains 10%FBS, according to requirement of experiment, be that 1.0 * 106cell/mL is inoculated in the culture plate of different apertures by cell density, 37 ℃ of incubators are hatched under 5% carbon dioxide conditions.Treat cell degree of converging to 70%, be changed to the DMEM culture fluid hunger that does not contain serum and spend the night that the serum-free medium that is changed to low sugar (5.5mmol/L) or high sugar (30mmol/L) then continues to hatch 24h.The HepG2 cell of hatching in low sugar culture-medium is the blank cell, and the HepG2 cell of hatching in high glucose medium is the IR-HepG2 cell.When setting up insulin resistant model, give the healht-care liquid of different hardness, observe healht-care liquid to the preventive effect of insulin resistant.When detecting the insulin signaling pathway correlative protein expression, before the harvesting, 100nM insulin stimulating 10min.
4) healht-care liquid is to the influence of IR-HepG2 cell sugar output: behind the drug incubation 24h, with sugar-free DMEM culture medium washed cell 3 times, add glucose output culture medium (sugar-free that contains 2mM Sodium Pyruvate and 20mM sodium lactate does not have phenol red DMEM) then and continue to hatch 16h, wherein last 3h adds the 1nM insulin.After hatching end, get supernatant and measure glucose content, BCA method mensuration protein content after lower floor's lysis in the plate.
5) healht-care liquid is to the influence of IR-HepG2 cell glycogen content: behind the drug incubation 24h, the PBS washed cell, add the culture medium that contains 1n M insulin and continue to hatch 3h, discard culture fluid, pre-cooling PBS washes 2 times, cell is scraped and is got, 12000r/min, 4 ℃ of centrifugal 15min abandon supernatant, add 0.5mL 30%KOH in the cell precipitation, put 100 ℃ of water-bath 20min, take out 10 μ L and measure protein content with the BCA method, all the other add the 1.5mL dehydrated alcohol, spend the night, the centrifugal 15min of 4000r/min abandons supernatant, adds distilled water 0.5mL in the precipitation, add 1mL 0.2% anthrone (the 0.2g anthrone is dissolved in 100mL 98% sulphuric acid) subsequently, 100 ℃ of water-bath 20min.Microplate reader detects the 620nm wavelength OD of place value, prepares the glucose solution of 100,50,25,12.5,6.25,3.125 and 1.56 μ g/mL simultaneously, does sugared concentration standard curve.
6) healht-care liquid is to the influence of IR-HepG2 cytolipin content: behind the drug incubation 24h, absorb remaining culture liq, ice PBS washes 2 times, add 100 μ L cell pyrolysis liquids, piping and druming was evenly incubated in the frozen water 30 minutes, constantly scrape with cell during this time and smear, lysate is fully contacted with cell; Lysate is transferred in the 1.5mL ep pipe low temperature ultrasonication 10s; 12000rpm, 15min, 4 ℃ are centrifugal ,-20 ℃ of freezing preservations, the cytolysate that takes a morsel is measured protein content with the BCA method, and all the other are for detection of intracellular lipid content.Measure the content of cell inner cholesterol and triglyceride respectively: unit calculates with every milligram of protein content (μ g/mg protein).
7) healht-care liquid is to the influence of ROS content in the IR-HepG2 cell born of the same parents: use the active oxygen detection kit to measure ROS content in the born of the same parents.Operate according to description, behind the drug effect 24h, the trypsinization harvesting, PBS washes 3 times, is resuspended in 10 μ M 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), hatch 30min for 37 ℃ then, ROS content in the fluorescent spectrophotometer assay born of the same parents, excitation wavelength 488nm, emission wavelength 525nm.
8) western blot detects healht-care liquid to the expression of associated protein such as insulin signaling pathway, AMPK signal path and JNK signal path and the influence of phosphorylation degree: cell pyrolysis liquid is measured protein concentration with the BCA method, sample on the equivalent, 10% separation gel SDS-PAGE, change film 1h, add IRS-2, pIRS-2 (Tyr612), Akt, pAkt (Ser473), GSK-3, G6Pase, AMPK, pAMPK, pHMGCR, pACC, SREBP-1 behind the 5%BSA sealing 2h, reach primary antibodie working solutions such as JNK, pJNK, actin, 4 ℃ of shaken over night.After primary antibodie was hatched, TBST washed film, added two anti-working solutions of the horseradish peroxidase-labeled of dilution in 1: 5000, and room temperature is shaken 1h, the ECL colour developing.
9) statistical procedures: data are all represented with mean ± standard deviation
Group difference adopts the t method of inspection to carry out.
2, experimental result
1) healht-care liquid is to the influence of HepG2 cell proliferation: as can be seen from Table 1, the healht-care liquid of different hardness does not have influence substantially to the propagation of HepG2 cell, therefore can be used for next step experiment.
Table 1: healht-care liquid is to the influence of HepG2 cell proliferation
2) healht-care liquid is to the influence of IR-HepG2 cell sugar output and glycogen content: it is the important special disease of insulin resistant that glycogen output increases, and glycogen output is made up of glyconeogenesis and glycogenolysis two parts.Can be learnt that by table 2 model group cell sugar output significantly increases, glycogen content then significantly reduces, and illustrates that high sugar processing makes hepatocyte that insulin resistant take place; And the processing of healht-care liquid can make the sugared output of IR-HepG2 cell significantly reduce, and glycogen content significantly increases, and illustrates that healht-care liquid can promote the synthetic of glycogen, thereby reduces glycogen output.
Table 2: healht-care liquid is to the influence of IR-HepG2 cell sugar output and glycogen content
Annotate: contrast with the blank group:
#P＜0.05,
##P＜0.01; Contrast with model group:
*P＜0.05,
*P＜0.01
3) healht-care liquid influence that IR-HepG2 cell IRS-2/PI3K signal related protein is expressed: Western blot result shows, as shown in table 3, behind the high concentration glucose effect 24h, the expressing quantity of IRS-2 is constant substantially, but the phosphorylation degree of IRS-2, Akt, GSK-3 significantly reduces, and the expression of G6Pase then significantly increases.Healht-care liquid then can dose dependent ground promotes the phosphorylation of IRS-2, Akt, GSK-3, suppresses the expression of G6Pase.Infer that healht-care liquid suppresses glyconeogenesis by this signal path, promote glycogen synthetic, improve insulin resistant thereby reduce glycogen output, specificity activates the IRS-2/PI3K signal path and is likely that healht-care liquid improves the target spot of insulin resistant.
Table 3: the influence that healht-care liquid is expressed IR-HepG2 cell IRS-2/PI3K signal related protein
Annotate: contrast with the blank group:
#P＜0.05,
##P＜0.01; Contrast with model group:
*P＜0.05,
*P＜0.01
4) healht-care liquid is to the influence of IR-HepG2 cell born of the same parents inner lipid content: as shown in table 4, behind the high concentration glucose effect 24h, model group cell born of the same parents inner lipid content is significantly higher than blank group, and the high sugar of prompting is handled and induced HepG2 lipid within endothelial cells deposition; Lipid content obviously reduces after 24 hours and process healht-care liquid is handled.
Table 4: healht-care liquid is to the influence of IR-HepG2 cell born of the same parents inner lipid content
Annotate: contrast with the blank group:
#P＜0.05,
##P＜0.01; Contrast with model group:
*P＜0.05,
*P＜0.01
5) healht-care liquid influence that IR-HepG2 cell AMPK signal related protein is expressed: western blot detects and finds that the AMPK phosphorylation level reduces in the model group cell, corresponding is the phosphorylation degree reduction of the synthetic rate-limiting enzyme HMGCR of cholesterol and fatty acid and ACC with it, the expression of SREBP-1 increases, and through after the healht-care water treatment, the AMPK activity significantly increases, the phosphorylation of HMGCR and ACC strengthens simultaneously, SREBP-1 expresses then corresponding reduction, the synthetic minimizing of lipids such as triglyceride and cholesterol in the final hepatocyte, insulin resistant improves, and these results show: health-care drinking-water can improve insulin resistant by the lipid route of synthesis that activates in the AMPK inhibition liver.
Table 5: the influence that healht-care liquid is expressed IR-HepG2 cell AMPK signal related protein
Annotate: contrast with the blank group:
#P＜0.05,
##P＜0.01; Contrast with model group:
*P＜0.05,
*P＜0.01
6) healht-care liquid to the IR-HepG2 cellular oxidation stress influence: as shown in table 6, high sugar is handled the phosphorylation degree that has significantly increased the interior ROS content of born of the same parents and JNK, the activity of important antioxidase GSH-Px then significantly reduces in the body, illustrate that redox equilibrium is seriously lacked of proper care in the model group cell, produced oxidative stress.After healht-care liquid was handled 24h, the phosphorylation degree of ROS content and JNK significantly reduced, and the activity of GSH-Px also obviously increases simultaneously, illustrates that this healht-care liquid can improve insulin resistant by the oxidation resistance of enhancing body.
Table 6: healht-care liquid to the IR-HepG2 cellular oxidation stress influence
Annotate: contrast with the blank group:
#P＜0.05,
##P＜0.01; Contrast with model group:
*P＜0.05,
*P＜0.01
This experimental result shows that this healht-care liquid can improve insulin signaling pathway preferably in external level, suppresses the excessively synthetic and oxidative stress of glyconeogenesis, lipid, effectively improves insulin resistant.This healht-care liquid can be used as the prevention that a kind of natural drink is used for insulin resistant and metabolic syndrome, metabolic syndrome patient can be according to health and the dietary habit healht-care liquid of selecting corresponding hardness of oneself, if diet is less to the picked-up of mineral, can select the healht-care liquid (600-1000ppm) of high rigidity for use, otherwise select the healht-care liquid (200-400ppm) than soft.
This healht-care liquid derives from deep seawater, has aboundresources, is easy to industrialization, and safe advantages of higher, has vast market and application prospect aspect the preventing and treating of hyperlipemia.The present invention will be the higher value application of deep seawater offer reference meaning and enlightenment.
Above embodiment is only in order to illustrating technical scheme of the present invention, but not limits it; Although with reference to previous embodiment the present invention is had been described in detail, for the person of ordinary skill of the art, still can make amendment to the technical scheme that previous embodiment is put down in writing, perhaps the special disease of part technology wherein is equal to replacement; And these modifications or replacement do not make the essence of appropriate technical solution break away from the spirit and scope of the present invention's technical scheme required for protection.
Claims (4)
1. a healht-care liquid that contains deep seawater is in preparation prevention or the medicine for the treatment of metabolic syndrome or the application in the health product, described healht-care liquid comprises A liquid and B liquid, described A liquid is fresh water, described B liquid makes through following steps: deep seawater is isolated dense water section behind reverse osmosis unit, dense water section is carried out concentrating under reduced pressure, make its volume be concentrated into the 0.67-3.33% of original volume, remove by filter the sodium chloride of separating out then, the height concentrated solution that obtains is B liquid, the hardness of described healht-care liquid is at 200-1000 ppm, and described deep seawater is the following and sea water after desalting processing of sea level 500m;
Magnesium in the described healht-care liquid: calcium: potassium: the concentration ratio of sodium element is 2.5-3.5:0.45-1:0.3-1:0.3-1.5, do not contain Organic substance, and water quality is alkalescence;
The water quality of described healht-care liquid is alkalescence, the pH value 7.5 ± 0.3 of 25 ℃ of mensuration;
Beneficial element content in the described healht-care liquid is: zinc 0.0325-16.4 μ g/L; Selenium 0.0395-0.712 μ g/L; Chromium 0.104-4.54 μ g/L; Manganese 0.156-5.52 μ g/L;
Harmful element content in the described healht-care liquid is: lead＜5 μ g/L; Hydrargyrum＜0. 5 μ g/L; Arsenic＜5 μ g/L; Cadmium＜1 μ g/L;
Described deep seawater is taken from apart from beyond the 20km of coastline.
2. a kind of healht-care liquid that contains deep seawater according to claim 1 is in preparation prevention or the medicine for the treatment of metabolic syndrome or the application in the health product, and its special disease is described magnesium: calcium: the ratio of greater inequality of the concentration of potassium is 3:1:1.
3. a kind of healht-care liquid that contains deep seawater according to claim 1 is characterized in that described fresh water sources Yu Haiyang deep water isolated fresh water after the reverse osmosis unit circulation in preparation prevention or the medicine for the treatment of metabolic syndrome or the application in the health product.
4. a kind of healht-care liquid of deep seawater that contains according to claim 1 is in preparation prevention or the medicine for the treatment of metabolic syndrome or the application in the health product, it is characterized in that it is the healht-care liquid of 600-1000 ppm that diet is selected hardness for use to the less metabolic syndrome patient of the picked-up of mineral, is the healht-care liquid of 200-400 ppm otherwise select hardness.
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| CN106211749A (en) * | 2014-03-31 | 2016-12-07 | 株式会社阿丽浱欧 | Comprise the functional drinks of the mineral water of the high rigidity made from salty subsoil water or deep seawater |
| CN108095111B (en) * | 2017-12-17 | 2021-06-18 | 锦上花医疗健康科技发展(广东)有限公司 | Nutritional composition containing deep seawater minerals and its application in promoting lactation after delivery |
| CN109602764A (en) * | 2018-05-14 | 2019-04-12 | 山东中医药大学 | A kind of application of health promoting liquid in preparation prevention and treatment Alzheimer disease drugs |
| CN111184737A (en) * | 2020-02-28 | 2020-05-22 | 山东中医药大学 | Application of fucoidan and deep seawater in prevention and treatment of diabetes cognitive impairment |
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