CN101209259A - Use of hyaluronic acid for chondrocyte oxidation resistance and hyperplasia - Google Patents
Use of hyaluronic acid for chondrocyte oxidation resistance and hyperplasia Download PDFInfo
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- CN101209259A CN101209259A CNA2006101723075A CN200610172307A CN101209259A CN 101209259 A CN101209259 A CN 101209259A CN A2006101723075 A CNA2006101723075 A CN A2006101723075A CN 200610172307 A CN200610172307 A CN 200610172307A CN 101209259 A CN101209259 A CN 101209259A
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Abstract
The invention relates to a usage of hyaluronic acid used for improving the oxidative damage of cartilage cells, so as to promote the proliferation of cartilage cells. More particularly, the invention relates to a usage of hyaluronic acid with the molecular weight of 0.6 to 1.2 million daltons used for protecting the activate oxygen damaged cartilage cells and promoting the proliferation of cartilage cells. The invention also relates to a usage of hyaluronic acid used for producing the pharmaceutical products for the treatment or prevention of arthritis and supplying the oral foods or drinks for the improvement of arthritis symptoms.
Description
Technical field
The present invention relates to the oxidative damage that hyaluronic acid (hyaluronic acid, have another name called nature of glass acid) is used to improve chondrocyte, and then promote the purposes of the proliferation of chondrocytes.The invention still further relates to hyaluronic acid and be used for making the confession treatment or prevent arthritic pharmaceuticals, and the purposes of the oral property Foods or drinks that improves arhritis conditions is provided.
Background technology
The joint provides the active hinge of organism skeleton.The two skeleton faces that intraarticular is adjacent cover flexible, the ganoid cartilage of one deck, and it has absorbing vibration and the effect of the power of reducing friction, to impel the movable smooth and easy of joint when skeleton actual contact and movable friction.Smooth and easy for the slip that increases the cartilage contact surface, intraarticular contains heavy-gravity joint fluid and has effect as lubricating oil.In cartilage and joint fluid, a kind of important component hyaluronic acid is arranged.Joint fluid contains the hyaluronic acid of the watery solution of 0.15% (w/v), mainly make by synovium of joint cell (synoviocyte), it is the poly-carbohydrate of a kind of macromolecular long-chain, its function is: 1. be present in the joint fluid with aqueous solution, provide viscosity (Pathophysiology.2003,9:215-220); 2. combine with some glucosaminoglycan molecules (Glucosaminoglycan, GAG etc.) and form Dan Baijutang (Proteoglycan), as one of main component of articular chondrocytes epimatrix; 3. the backbone who in cartilage, provides chondrocyte to adhere to and survive; 4. toxicity that can oxidated material and destroy its structure (referring to, Arthritis ﹠amp for example; Rheumatism.2003,48:3151-3158), and wherein last comes into one's own at present just day by day.
Because the mechanicalness loss of joint motions or the aging other factors that waits can reduce metabolic oxidant and increase oxide or free radical etc., and cause the injury of chondrocyte.Owing to contain hyaluronic acid in the cartilage matrix, it must keep constant structure (matter) and amount by sound chondrocyte, just can be for keeping normal cartilage structure.But these cells in the mid-term in human life cycle or more early, even aging apoptosis gradually just becomes and just formed so move back the shape sexually transmitted disease (STD).The reason of chondrocyte deterioration is very complicated, few known to the educational circles at present, but generally acknowledged key factor, be exactly the oxidizing substance, the particularly injury that it is caused as strong oxidizers such as free radicals that produces in the metabolic process (comprise aforementioned mechanicalness abrasion and give birth to biological running of body nature or the like).
In chemical reaction, active oxygen (reactive oxygen species, ROS) or be called oxygen-derived free radicals (freeradicals) can exist with unsettled situation, must obtain an electronics and reach balance, therefore can be in the mode of oxidation, with other atom or chemical compound competition electronics.Have approximately in the interior all free radicals of human body according to estimates to belong to ROS more than 95%, the active oxygen or derivatives thereof can influence the state of other molecule or chemical compound because of the electronics of vying each other.Many places all can produce free radical in the structure of cell: as mitochondrion, lysosome, cell membrane phospholipid matter, endoplasmic reticulum etc.Free radical is too much understood the pair cell tissue and is damaged.When intravital oxide is too much, can cause intravital dysequilibrium, and cause lipid, protein or DNA damage, and then make such as diseases such as cancer, Parkinson's disease, hypertension, arteriosclerosis, myocardial infarction, thrombosis, platelet aggregations and produce.The unsaturated bond combination of lipid produces lipid peroxidation, and makes damaged membrane on present known free radical meeting and the cell membrane.Free radical also can damage and contain sulfoprotein and other albumen, causes ion transport system (ion transport systems) impaired, causes cell to come to harm or causes protein decomposition, structural change or degeneration etc.Free radical can damage the base of DNA (deoxyribonucleic acid) in addition, makes dna break, sudden change, cell cycle change even has carcinogenecity.
At present known in arthritis, active oxygen is (as O
2 -, OH
, H
2O
2, HOCl etc.) can carry out depolimerize (depolarization) and destroy the hyaluronic acid structure, and reduce hyaluronic quantity, the viscosity that reduces the joint and lubricity and generation inflammation phenomenon (referring to, Free Radic Biol Med.2003 for example, 35:169-78; And Pathophysiology.2003,9:215-220).H
2O
2And CuCl
2Existence can destroy the hyaluronic structure of high molecular (for example 1,200,000 dalton) (referring to Carbohydr Res.1999,321:228-34; Carbohydr Res.2006,341:639-44; And Pathophysiology.2003,9:215-220).Existing document mentions that also the hyaluronic acid in the joint fluid is subject to H
2O
2Or by H
2O
2Deutero-OH
Destroy (Inflammation.1993,17:403-15).Therefore, H
2O
2Appear as the main cause that causes the hyaluronic acid structural change.The present invention utilizes different molecular weight and the hyaluronic acid of making the source, assesses hyaluronic acid as an antioxidation (H at external or live test
2O
2), and and then promote the purposes of the proliferation of chondrocytes.And assessment add hyaluronic acid can be in the process that the person joint degenerates, provide to a certain degree reduction and the function of protection cell, or have the effect of the proliferation of chondrocytes.
Summary of the invention
The object of the invention and advantage are described in down part, or can be by apparent in describing.
The object of the present invention is to provide hyaluronic acid (hyaluronic acid) to be used to improve the purposes of chondrocyte because of the caused oxidative damage of active oxygen.
Another object of the present invention is to provide hyaluronic acid to be used to promote the purposes of the proliferation of chondrocytes.
Above-mentioned purpose of the present invention is achieved in that a kind of purposes of hyaluronic promotion the proliferation of chondrocytes, is to realize by the regulation and control to the chondrocyte growth cycle.
Purposes of the present invention, the regulation and control of this growth cycle are to increase the performance of growth cycle regulatory factor CyclinB1 in the G2/M phase.
Another purpose of the present invention is to be provided for treatment or prevents arthritic pharmaceuticals.
Above-mentioned purpose of the present invention is achieved in that and a kind ofly is used for the treatment of or prevents arthritic pharmaceuticals, it is characterized in that comprising the hyaluronic acid of molecular weight between 50-800 ten thousand dalton (Dalton).
Pharmaceuticals of the present invention, this hyaluronic acid is used to improve the oxidative damage of articular chondrocytes.
Pharmaceuticals of the present invention, this hyaluronic acid are used to reduce the free radical activity in the joint fluid.
A further object of the present invention is to be provided for improving the oral property food of arhritis conditions.
Above-mentioned purpose of the present invention is achieved in that a kind of confession improves the oral property food of arhritis conditions, it is characterized in that, comprises molecular weight between 50-800 ten thousand daltonian hyaluronic acids.
Below, in conjunction with specific embodiments and shown in accompanying drawing, the present invention is described in further detail.
Description of drawings
Fig. 1 represents that the hyaluronic acid (Hyaluronan) of various different molecular weights reduces H in the joint fluid
2O
2Generation.
After Fig. 2 is illustrated in and gives patient's hyaluronic acid, the O in its joint fluid
2 -, H
2O
2, and the inflammatory factor c reactive protein (C-reactive protein is CRP) with the amount of hoptoglobin (haptoglobin), than the decline situation of matched group.
Fig. 3 A is illustrated in and gives under the hyaluronic environment, and the performance of free radical is suppressed.
Fig. 3 B has been illustrated in or has not had hyaluronic acid (20 μ l) and existed down, and chondrocyte is along with H
2O
2The survival rate that concentration increases.
Fig. 4 has been illustrated in or has not given under the hyaluronic situation, old human chondrocyte growth situation in time.
Fig. 5 represents to utilize flow cytometer (flow cytometery) analysis to have or the cell cycle of old man's chondrocyte of handling without hyaluronic acid (1mg/ml).
Fig. 6 represents with the western blot analysis protein of regulation and control G0/G1 phase: cyclin D1, cdk4 and cdk6; And the protein of regulation and control G2/M: cell periodic protein B 1, the performance in the chondrocyte that has or handle without hyaluronic acid (1mg/ml) changes situation.
The specific embodiment
Following examples are for using Japan Patent: the isolating hyaluronic acid goods of 1284023 and 1353027 mode.Wherein comprise: 6,000,000 dalton's hyaluronic acid (Synvisc, Biomatrix, USA), 60-120 ten thousand dalton's hyaluronic acid (ARTZDispo, Seikagaku, Japan), 60-120 ten thousand dalton's hyaluronic acids (Hikamilon Dispo, Taisho Pharmaceutical Co.Ltd.Japan), 60-120 ten thousand dalton's hyaluronic acid (Lumisteron Dispo, Nissin, Japan), 60-120 ten thousand dalton's hyaluronic acid (UnihylonDispo, UJI, Japan), 50-73 ten thousand dalton's hyaluronic acid (Hyalgan, Fidia, Italy), 50-73 ten thousand dalton's hyaluronic acids (Suplasyn, Bioniche, Ireland).
Embodiment
Use hyaluronic acid to flow into patient's intraarticular clinically for understanding, to the influence of free radical performance in the joint fluid how it at first take out the patient's who was subjected to the hyaluronic acid treatment joint fluid, detects contained free base unit weight in it.
Collected human joint's liquid is placed on ice.The joint fluid of getting 200 μ l places in the iron pan, and puts in the detection groove of chemical luminescence detection instrument (CLA-FS1, Tohoku Electronic Ind.Co.Sendai, Japan).The start detection instrument, after surveying one section background value in 50 seconds, add 500 μ l luminol (Luminol (5-amino-2 again, 3-dihydro-1,4-phthalazinedione), buy Sigma company in the U.S., powder be dissolved in that phosphate buffer normal saline (PBS) is mixed with 0.1mM and in 4 ℃ of preservations) or lucigenin (Lucigenin (Bis-N-Methylacridinium), buy Sigma company in the U.S., powder be dissolved in that phosphate buffer normal saline (PBS) is mixed with 0.1mM and in 4 ℃ of preservations) and then the content that detects free radical stopped (can getting a stored counts value in per 10 seconds) up to 300 seconds.Storage file is also analyzed.The computation time-integral mean of 50 seconds gained of counting, be multiplied by 30 background integrated values that can get 300 seconds again.With 300 seconds the background integrated value of area integral value deduction that all time-counting constituted, remove last 25 again and can obtain the per 10 seconds average free radical counting of a corpse or other object for laboratory examination and chemical testing.The result is shown in Fig. 1.Show that by Fig. 1 50-800 ten thousand daltonian hyaluronic acids have H in the joint fluid of reduction
2O
2The effect of generation.Wherein reduce H
2O
2Expressive ability be the daltonian hyaluronic acids in 60-120 ten thousand daltonian hyaluronic acid＞50-73 ten thousand daltonian hyaluronic acid＞6,000,000.
Fig. 1 represents that the hyaluronic acid (Hyaluronan) of various different molecular weights reduces H in the joint fluid
2O
2Generation.Wherein reduce H
2O
2Expressive ability be 60-120 ten thousand dalton＞50-73 ten thousand dalton＞6,000,000 dalton.
Joint fluid with the patient that do not accept hyaluronic acid treatment is a matched group, is shown by experimental result, in the joint fluid of matched group, contains a large amount of O
2 -, H
2O
2Free radical, yet after giving patient's hyaluronic acid, the H in its joint fluid
2O
2Amount is than the tangible decline of matched group (referring to Fig. 2).
For understanding in experiment in vitro, whether hyaluronic acid can lower free radical activity, gets the H of 200 μ l variable concentrations
2O
2(0,100,200 and 400 μ M) utilizes chemical cold light analyzer to measure the performance situation of its free radical.And the other performance amount that under the environment of 200 μ M, gives the hyaluronic acid (hyaluronic acid) of 20 μ l and measure free radical.The result as shown in Figure 3A, the performance amount of free radical can be along with H
2O
2Concentration increase and increase, but at H
2O
2200 μ M have and add under the acid-treated environment of 20 μ l hyalomitomes, find that the performance of free radical is suppressed, and almost and not add H
2O
2The time performance the same.Owing to learn from document, the content of free radical increases, and can cause the death of cell.So, whether can reduce the cell mortality that causes because of free radical for inquiring into hyaluronic acid (hyaluronic acid), collect elderly patients' (greater than 60 years old) cartilaginous tissue, and then isolate primary chondrocyte and cultivate.
To the centrifuge tube weighing record of a corpse or other object for laboratory examination and chemical testing be housed, deduct the nt wt net weight of centrifuge tube, the weight of a record cartilage corpse or other object for laboratory examination and chemical testing.Add 10ml phosphate buffer normal saline (PBS) in centrifuge tube, horizontally rotate and rock five times, again PBS is blotted.A repeated washing cartilage corpse or other object for laboratory examination and chemical testing once.Utilize flat mouth forceps to pick up a cartilage corpse or other object for laboratory examination and chemical testing, be transferred on the 10cm Tissue Culture Dish.With 10ml PBS a cartilage corpse or other object for laboratory examination and chemical testing is cleaned once again.Utilize scalpel that a cartilage corpse or other object for laboratory examination and chemical testing is done initial gross separation, will adhere to the soft tissue and the os osseum separate tissue of a corpse or other object for laboratory examination and chemical testing; Isolated lily cartilaginous tissue is moved on in the sterile petri dish of step 8.(volume is no more than 1mm to utilize scalpel that fractionlet is cut in the cartilaginous tissue segmentation
3).Add the plain catabolic enzyme (100mg/ml) of 60 μ l, second fiber type.The Tissue Culture Dish that is equipped with segmentation back cartilaginous tissue is moved placement 37 ℃, 5%CO
2Shaker in the cell culture incubator rocked four hours with 50rpm.Draw backwash cartilaginous tissue after decomposing of 5ml cell culture fluid, all move in the preliminary aseptic 15ml centrifuge tube.Draw the 5ml cell culture fluid again and backwash that it decomposes the culture dish of cartilage, all collect in the preliminary aseptic 15ml centrifuge tube to guarantee the cell that utilizes the plain catabolic enzyme of second fiber type to separate.Under room temperature, centrifugal 1000rpm 5 minutes.Sop up supernatant, add 10ml PBS and break up cell, recentrifuge 1000rpm 5 minutes.Repeated washing chondrocyte Two time.Add the 10ml cell culture fluid and evenly break up cell, get its 50 μ l cell culture fluid then and move on to the centrifugal tubule of 1.5ml, 10 * trypan blue the dyestuff (Trypan Blue) that adds 40 μ l PBS and 10 μ l is got 10 μ l mixed liquors and is slowly put blood cell calculator into and count at microscopically behind the uniform mixing.The cell density of counting: the count number of nine lattice * 2 * 104/9=cell number/ml.Chondrocyte changes the implantation toper bottle with 1 * 104/ml cell density, and is placed in 37 ℃, 5%CO
2Cell culture incubator.Utilized deserted suction pipe to sop up cell culture fluid in the conical flask in per three days, add 10ml PBS in taper Ping Li face upper limb (being sure not directly to impact cell), after level relaxes and rocks conical flask, sop up phosphate buffer normal saline (phosphate buffer saline), add the 12ml cell culture fluid again in conical flask, and conical flask is continued to be put in 37 ℃, 5%CO
2Cell culture incubator is cultivated.
When surviving experiment, in culture medium, add the H of variable concentrations
2O
2(0,100,200 and 400 μ M), other two groups is the H of 0 and 200 μ M
2O
2All add 20 μ l hyaluronic acids, so cultured cell was collected chondrocyte and is calculated its survival rate after one day.Show that from the result who is shown in Fig. 3 B the survival rate of chondrocyte is along with H
2O
2Concentration increase and descend, but containing under the hyaluronic environment, the survival rate of cell increases on the contrary.Therefore, prove that hyaluronic acid has the free radical of reduction performance amount really, and reduce the mortality rate of chondrocyte, and then promote the effect of the proliferation of chondrocytes.
Fig. 3 B has been illustrated in or has not had hyaluronic acid (20 μ l) and existed down, and chondrocyte is along with H
2O
2The survival rate that concentration increases.The result shows that the survival rate of chondrocyte is along with H
2O
2Concentration increase and descend, but increase on the contrary in the survival rate that cell under the hyaluronic environment is arranged.
Embodiment 3. hyaluronic acids promote old people's chondrocyte is grown
This experiment further test to the influence that the chondrocyte in old man joint is grown, is measured the growth curve of chondrocyte in the presence of having or not having hyaluronic acid.According to the step of previous embodiment 2, cultivate old man's chondrocyte, and in cell grow to about eighty per cant when full with its branch dish, every dish about 8 * 104/60mm dish also is cultured to overnight.Every other day, take out the cell culture dish, cell culture fluid is replaced as the cell culture fluid that contains 1mg/ml hyaluronic acid (hyaluronic acid) comes cultured cell (HA group), matched group is then changed general culture fluid, till incubation time continues to 12 days, and in culture fluid of replacing in the 5th day.Calculate total cellular score since the 0th day (Day0), calculated once in then per two days.This tests triplicate.The result is shown in Fig. 4.By measured growth curve as can be known, old human chondrocyte is having under the situation that gives hyaluronic acid (hyaluronic acid), can find on the from the 2nd to the 4th day that the cell number of HA group was significantly greater than 1.7 times of matched groups, and reach difference than matched group greater than twice the 6th day HA group.The HA group speed of growth eased up in the 8th to 12 day, but compared with matched group, still had 1.4 to 1.6 times difference.
Embodiment 4. hyaluronic acids are regulation and control via growth cycle to old people's chondrocyte growth
Whether can regulate and control the cell cycle of old human chondrocyte for understanding hyaluronic acid, step according to previous embodiment 2, in eighty per cant when full with its branch dish, about 3 * 105/100mm the dish of every dish also contains the hyaluronic cell culture fluid of 1mg/ml and comes cultured cell (HA group) in every other day cell culture fluid being replaced as, matched group is then changed general culture fluid, till incubation time continues to 8 days.Since the 0th day (Day0) collecting cell, collected once in then per two days.Collected cell, treated after, utilize flow cytometer (flow cytometery) to analyze its cell cycle.
Prepare single-cell suspension liquid earlier., handle with Trypsin matter enzyme again with the PBS rinse through cultured cells, treat cell rounding after, just the culture fluid that contains hyclone (5%FCS) with 5ml is collected cell, and breaks up cell suspending liquid equably.Add the ice-cold PBS of 5ml and clean cell, low-speed centrifugal 5 minutes is removed supernatant afterwards carefully, can stay about 50 μ l solution, avoids brushing up against cell.Add 0.2ml PBS and break up cell equably.The cell mixing suspension that vibrates on one side simultaneously drop by drop adds 1ml 70% ethanol with fixed cell, rests in-20 ℃ of refrigerators at least one hour (but after the alcohol fixation sample standing over night).With centrifugal 5 minutes of low speed (1200 rev/mins), absorb supernatant and avoid touching cell.Cell mass is broken up equably, added 5ml PBS and clean cell.Leave standstill after one minute low speed 1200rpm centrifugal 5 minutes, and absorbed supernatant afterwards as far as possible.Add 1ml propidium iodide PI/ NONIN HS 240 TritonX-100 dyeing liquor, cell mass is broken up equably, and jog mixes, (ultimate density is a NONIN HS 240 (Triton X-100) 0.1%, ribonucleic enzyme A (RNase A 0.2mg/ml ﹠amp at room temperature to act on 30 minutes in the darkroom; Propidium iodide PI 20 μ g/ml avoid reusing the enzyme of rising again).Mixing sample before the machine on facing, and with 35-μ m nylon screen filtered sample.In two hours of sample preparation, utilize flow cytometer (flow cytometer) (FL2-A) to detect the performance of cell fluorescence, and can be according to the variation of the data analysis cell cycle of gained.
The result is shown in Fig. 5.In the measurement result (referring to Fig. 5 A) of G0/G1 phase, the cell number percentage ratio HA of matched group group is high in the time of the 4th day, and all the other are zero difference then.And, between two groups of experimental group and matched groups, do not have statistical difference on the from the 0th to 8 day in the measurement result (referring to Fig. 5 B) of S phase.In addition, in the measurement result (referring to Fig. 5 C) of G2/M phase, the cell number percentage ratio of HA group but did not then have difference on the 6th day later on than the matched group height when finding the 4th day.
For further understanding the carrying out how hyaluronic acid changes cell cycle, whether factor-related with cell cycle regulating, step according to previous embodiment 2 is cultivated old man's chondrocyte, and in having or not giving under the hyaluronic environment of 1mg/ml, cultivated 2,4,6 and 8 days, and behind the protein of collecting cell, analyze the performance situation of the wherein various different cell cycle regulating factors with western blotting (Western blotting).
Clean three cells with 1 times of PBS, and adding RIPA buffer (50mM Tris (pH7.5), 150mM NaCl, 1%NP40,0.1% sodium lauryl sulphate SDS, 0.5% NaTDC (sodiumdeoxycholic acid), protease inhibitor (proteinase inhibitor)) cell is broken and stripping protein.Be used spatula, cell is scraped from culture plate, be collected into microcentrifugal tube.Centrifugal (12000 rotating speeds, 30min, 4 ℃) take out the supernatant preparation and carry out electrophoretic separation.A protein quantification corpse or other object for laboratory examination and chemical testing later takes out 20 μ g, and according to sample: the mixed of sample buffer (containing glycerol)=1: 4, and heating five minutes under 100 ℃ water temperature, application of sample is to electrophoresis tank.Carry out electrophoretic after, take out colloid and cut the nylon paper that size is fit to, ((Tris 30.28g and glycine (glycine) 144.9g is configured to one liter to the 100ml concentrated solution to immerse transfering buffering liquid, pH 8.2～8.4)+700ml redistilled water+200ml methanol) in, paper is covered on the colloid, put into diversion box (transfer chamber) together, connect electrode and adjust electric current and open.Beginning shifts.The condition that shifts (Transfer) is: 300mA, 2.5～3hr.After shift finishing, nylon paper is taken out and place 5% skim milk to be dissolved in the Tris buffer, under room temperature, shook one hour.Nylon paper is cleaned up (cleaning each five minutes three times) with the Tris buffer, add that one-level antibody shook one hour or shake under room temperature and spend the night in 4 ℃.With Tris buffer solution for cleaning nylon paper, clean each five minutes five times.The adding secondary antibody was also shaken under room temperature 40 minutes.Use Tris buffer solution for cleaning nylon paper as described above, back adding chemiluminescent assay group (ECL kit) (solvent 1: solvent 2=1: 1), solution need be covered in nylon paper fully, at room temperature reacts one minute.Develop to dark place.
The result as shown in Figure 6, regulation and control the G0/G1 phase protein as: cyclin D1, cdk4 and cdk6 all do not have difference in matched group and hyaluronic acid processed group (HA group).But in the chondrocyte of handling hyaluronic acid (hyaluronic acid), in the time of the 4th day, the protein of regulation and control G2/M phase: cell periodic protein B 1 performance amount is a little more than matched group.Then do not change with cell periodic protein B 1 relevant protein cdc2 in addition.
In sum, the present invention has utilized the hydrogen peroxide (H of variable concentrations
2O
2) be added in the culture environment of chondrocyte, come analog cell to suffer the situation of oxidation material infringement.In these systems, add the hyaluronic acid in different molecular weight or source then, observe whether can lower cytogenetic damage.The front of this research is found, can further specify the hyaluronic effective machine commentaries on classics of injection in articular cavity, provides control arthritic another kind of effective model simultaneously.Of the present invention in addition resisting-H
2O
2Hyaluronic acid also is applicable to because of the hemochromatosis of ferrum excessive accumulation, bronze diabetes and pigmentary cirrhosis.Based on above experimental result, the present invention also proposes to contain hyaluronic acid and is used to make the purposes that confession improves the oral property Foods or drinks of arhritis conditions.
Claims (15)
1. one kind is used to improve the purposes of the oxidative damage of chondrocyte with hyaluronic acid, and wherein this oxidative damage is caused by active oxygen.
2. purposes according to claim 1, wherein this hyaluronic molecular weight is between 50-800 ten thousand dalton.
3. purposes according to claim 2, wherein this hyaluronic molecular weight is between 60-120 ten thousand dalton.
4. purposes according to claim 1, wherein this active oxygen is selected from O
2 -, OH
, H
2O
2And HOCl.
5. purposes according to claim 1, wherein this oxidative damage that improves chondrocyte is to realize by the free radical activity that lowers active oxygen.
6. purposes according to claim 1, wherein hyaluronic acid can further reduce the mortality rate of chondrocyte.
7. purposes that hyaluronic acid is used to promote the proliferation of chondrocytes.
8. purposes according to claim 7, wherein hyaluronic acid promotes that the proliferation of chondrocytes is to realize by the regulation and control to the chondrocyte growth cycle.
9. purposes according to claim 8, wherein the regulation and control of this growth cycle are to increase the performance of growth cycle regulatory factor Cyclin B1 in the G2/M phase.
10. one kind is used for the treatment of or prevents arthritic pharmaceuticals, it is characterized in that, comprises molecular weight between 50-800 ten thousand daltonian hyaluronic acids.
11. pharmaceuticals according to claim 10, wherein this hyaluronic molecular weight is between 60-120 ten thousand dalton.
12. pharmaceuticals according to claim 10, wherein this hyaluronic acid is used to improve the oxidative damage of articular chondrocytes.
13. pharmaceuticals according to claim 10, wherein this hyaluronic acid is used to reduce the free radical activity in the joint fluid.
14. a confession improves the oral property food of arhritis conditions, it is characterized in that, comprises molecular weight between 50-800 ten thousand daltonian hyaluronic acids.
15. food according to claim 10, wherein this hyaluronic molecular weight is between 60-120 ten thousand dalton.
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| CN112972490A (en) * | 2021-03-04 | 2021-06-18 | 中国人民解放军军事科学院军事医学研究院 | Application of hyaluronic acid in preparing medicine for preventing or treating iron death related diseases |
| WO2022184091A1 (en) * | 2021-03-04 | 2022-09-09 | 中国人民解放军军事科学院军事医学研究院 | Application of hyaluronic acid in preparation of drugs for prevention or treatment of diseases related to ferroptosis |
| CN114081114A (en) * | 2021-11-12 | 2022-02-25 | 华中农业大学 | Sodium hyaluronate beverage with function of regulating intestinal flora and preparation method thereof |
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