CN102273637B - Application of monascus mycelium extracts in preparation of health-care food and medicines for preventing osteoporosis - Google Patents
Application of monascus mycelium extracts in preparation of health-care food and medicines for preventing osteoporosis Download PDFInfo
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Abstract
The invention relates to application of monascus mycelium extracts left after haematochrome is extracted by a monascus liquid state fermentation process to the preparation of health-care food and medicines for preventing osteoporosis. Researches prove that raw materials such as rice and the like are subjected to submerged fermentation of liquid by monascus to generate a monascus fermentation product, and after monascus haematochrome is extracted from the monascus fermentation product by 60 to 70 percent ethanol, the residual monascus mycelium extracts subjected to acid treatment or alkali treatment have the effects of reducing blood fat and preventing the osteoporosis, and can be used as medicines or food additives to be used for preparing the medicines or the health-care food for preventing the osteoporosis.
Description
Technical field the present invention relates to adopt the red colouring agent for food, also used as a Chinese medicine liquid-state fermentation technology to extract the application of the extract of the Monascus anka Nakazawa et sato filament that is left behind the haematochrome at preparation prevention of osteoporosis health food and pharmaceutical preparations.
The background technology osteoporosis is the modal disease of the elderly, and its incidence of disease occupies common disease, frequently-occurring disease the 7th, and about 200,000,000 people in the whole world suffer from osteoporosis.In the U.S. and western countries, in the age surpasses 55 years old postclimacteric women, there is half the people to suffer from the different osteoporosis of degree, and in the elderly of over-65s, suffers from osteoporosis person up to more than 95%.Along with the reach of science and human progress, the average life span constantly prolongs, and elderly population increase sharply, and the osteoporotic incidence of disease is also along with increase rapidly occurring.Show that according to more American-European national statisticses the incidence of disease of fracture increased by 40%～60% in per 10 years approximately at present.Estimate to the year two thousand fifty whole world over-65s the elderly will be increased to 15.55 hundred million by present 3.23 hundred million; The number of hip fracture generation when the time comes also will be increased to 6,260,000 by present 1,660,000, and wherein the patient of Asia, Latin America, the Middle East and African country will account for more than 70%.China is the maximum country of world population, wherein elderly population account for the Asia elderly population 1/2 with world population 1/5.The osteoporosis epidemiology survey result in Beijing, Shanghai and 3 cities, Chengdu shows, the 60-69 year osteoporotic incidence women of age bracket and the male sex are respectively about 60% and 30%.China's large population base, sufferers of osteoporosis face estimates to have reached 6,000 ten thousand～8,000 ten thousand examples, this brings serious burden for family and society.People according to another statistics national over-65s in 2000 has 1.3 hundred million, accounts for 10.7% of national population, and it is national to have become typical veteran form.And the patient who suffers from osteoporosis accounts for 90% in these crowds, and the patient of hip fracture will have 12%～40% to die from various complication in 1 year.50% human action inconvenience is also arranged in the survivor.This has not only caused serious body and mind to wreck to patient, has increased huge manpower and financial resources burden also for simultaneously family and society.Therefore, it is anti-ageing preventing and treating osteoporosis, life-saving, a research topic that ten minutes is urgent of the assurance people's quality of life.
The China that is applied in of monascus has history of more than one thousand years, and it is the main bacteria seed of producing wine of rice fermented with red yeast, red yeast rice, functional Monascus and natural colouring matter.In recent years, Along with people's is to synthesizing food additives and the chemicals understanding to the human body toxic and side effect, and countries in the world have started edible pollution-free food and the upsurge of using natural drug, and red colouring agent for food, also used as a Chinese medicine has caused great concern as the natural product of dietotherapeutic.From 1979; Japan scholar rattan chapter far away is found the Lovastatin physiological activator from monascus (monaseusruber) after; Cause whole world scientist's concern; 1980～nineteen ninety peaks for the red colouring agent for food, also used as a Chinese medicine functional study, and function such as the reducing blood lipid of red colouring agent for food, also used as a Chinese medicine, norcholesterol, hypotensive, hypoglycemic, antibiotic property comes to light in succession.Merck company succeeds in developing first and is used for clinical Lovastatin lipid lowering agent, and in listing in 1987, oneself became high fat of blood, coronary disease patient's choice drug present statins, and lipid lowerers has become second-biggest-in-the-world medicine.The present production technology of red colouring agent for food, also used as a Chinese medicine mainly is to design for extracting monascorubin, generally all is that the selection high quality white rice is a raw material, adopts modern biotechnology to isolate the monascus ruber of high-quality; Through liquid deep layer fermenting, produce the red koji fermentation product again, these products are through dehydration; Technology such as press dry; With 60～70% alcohol extract haematochrome, usually behind these extraction process purification haematochrome in the red colouring agent for food, also used as a Chinese medicine component for reducing blood fat Lovastatin also withdraw together, the Lovastatin that remaining slag (Monascus anka Nakazawa et sato filament) stays seldom or does not almost have; Can produce a large amount of Monascus anka Nakazawa et sato filament slags in this production process; These Monascus anka Nakazawa et sato filament slags generally are regarded as useless discarded object and discard, and this has not only caused the pollution of environment, especially to the significant wastage of resource.We find in the detection of two batches of Monascus anka Nakazawa et sato filaments that sky, Dongguan benefit biotechnology company is produced, though its discarded Monascus anka Nakazawa et sato filament finds the Lovastatin that can the specificity antagonism suppresses the HMC-CoA reductase activity, also have aliphatic acid and polysaccharide composition in the Monascus anka Nakazawa et sato filament extract; Adopt these discarded Monascus anka Nakazawa et sato filaments; After peracid treatment or alkali treatment, the Monascus anka Nakazawa et sato filament extract that extracts carries out the zoopery of reducing blood lipid and prevention of osteoporosis, and unexpected these Monascus anka Nakazawa et sato filament extracts of finding also have the effect of good effect for reducing blood fat and prevention of osteoporosis; Our these Monascus anka Nakazawa et sato filament extracts of discovering not only have effect and the effect of increasing high density lipoprotein of the reduction serum cholesterol of highly significant; And the osteoporosis that hyperlipidemia is brought out had tangible prevention effect, and pointed out in these Monascus anka Nakazawa et sato filaments that extracted monascorubin, must also have some compositions that osteoporosis is had prevention effect; These red colouring agent for food, also used as a Chinese medicine Monascus anka Nakazawa et sato filament extracts should be a kind of extraordinary medicine or the food additives of prevention of osteoporosis; After the further purification of these Monascus anka Nakazawa et sato filament extracts,, make tablet by the traditional Chinese medicines production technology; Capsule; Granule, electuary etc. can supply the pharmaceutical dosage form of clinical practice, can supply to suffer from patient's clinical practice of osteoporosis disease.In addition; Add these Monascus anka Nakazawa et sato filament extracts in the biscuit raw material; The biscuit of producing carries out the zoopery of reducing blood lipid and prevention of osteoporosis; Proved that also these biscuits have the effect of tangible reducing blood lipid and prevention of osteoporosis, can be used as a kind of good health care food of prevention of osteoporosis.
Summary of the invention invention adopt raw material such as rice with monascus behind liquid deep layer fermenting; The red koji fermentation product that produces; After with 60～70% alcohol extract monascorubins; The effect that the extract of remaining monascus ruber filament after acid treatment or alkali treatment has reducing blood lipid and prevention of osteoporosis can be used as medicine or food additives, is used to prepare the medicine or the health food of prevention of osteoporosis.
The Monascus anka Nakazawa et sato filament extract that the present invention mentions can prepare through following method:
(1) get raw material such as rice with monascus through liquid deep layer fermenting after 7 to 10 days, the product liquid of the monascus of generation fermentation removes water partial pressure through squeezer, then; 60～70% alcohol extract haematochrome have extracted the Monascus anka Nakazawa et sato filament of haematochrome, dry; Press per kilogram and add 0.1～0.5 liter of 1% acetic acid, under 25 ℃ of room temperatures, keep more than 24 hours, by gross weight 5～8 extraordinarily water extract; Extract twice, each 1 hour, reclaim extracted twice liquid; The dried cream of simmer down to is pulverized and is fine powder, room temperature preservation;
(2) get raw material such as rice with monascus through liquid deep layer fermenting after 7 to 10 days, the product liquid of the monascus of generation fermentation removes water partial pressure through squeezer, then; 60～70% alcohol extract haematochrome have extracted the Monascus anka Nakazawa et sato filament of haematochrome, dry; Press per kilogram and add 0.1～0.5 liter of 1% sodium acid carbonate, under 25 ℃ of room temperatures, keep more than 24 hours, by gross weight 5～8 extraordinarily water extract; Extract twice, each 1 hour, reclaim extracted twice liquid; The dried cream of simmer down to is pulverized and is fine powder, room temperature preservation;
(3) get raw material such as rice with monascus through liquid deep layer fermenting after 7 to 10 days, the product liquid of the monascus of generation fermentation removes water partial pressure through squeezer; Then, 60～70% alcohol extract haematochrome have extracted the Monascus anka Nakazawa et sato filament of haematochrome; Dry,, extract twice by extraordinarily water extraction of gross weight 5～8; Each 1 hour, reclaim extracted twice liquid.The dried cream of simmer down to is pulverized and is fine powder, room temperature preservation.
The monascus hypha extract that the present invention prepares as stated above can be made tablet by the traditional Chinese medicines production technology, capsule, and granule, electuaries etc. can supply the pharmaceutical dosage form of clinical practice, supply to suffer from patient's clinical practice of osteoporosis disease.
The monascus hypha extract that the present invention extracts as stated above can be used as the biscuit additive and adds in the factory formula of biscuit, produces the biscuit or the health food that contain the red colouring agent for food, also used as a Chinese medicine slag, and supplying has the patient who suffers from the osteoporosis disease to use as healthy food.In 5～30% scopes, is good with 10～20% at the addition of biscuit product.
The red rice residue product that the present invention extracts as stated above can be used as beverage additive and adds in the beverage formula, produces the beverage that contains Red Yeast Rice P.E, and supplying has the patient who suffers from the osteoporosis disease to use as healthy food.In 5～30% scopes, is good with 10～20% at the addition of biscuit product.
The specific embodiment:
Embodiment one:
1 materials and methods
1.1 animal used as test and raising condition
62 of 3 monthly age of SPF level male SD rats, body weight (251.8 ± 18.3) g is provided by Guangdong Medical Lab Animal Center, the medical experiment animal quality certification number: word 2008A020 checks and affirm in Guangdong.
The raising condition: indoor temperature keeps 24 ℃～28 ℃ during raising, and humidity is 50%～60%, special chamber sub-cage rearing, the next day change bedding and padding, freely take the photograph water, ingest (standard feed nursing) weighs once weekly.
1.2 experimental design and grouping
Choose 62 of 3 monthly ages cleaning level male SD rats, the body weight of rat used Microsoft Excel produce random number series, and utilize the random number series that produces to be divided into 7 groups (seeing table 1.1) at random:
Table 1.1 animal used as test divides into groups and dosage
Divide group profile:
The BAS group: the basal year level control group, tested 0 day, do not give usefulness, by the experimental technique taking-up detection of back, do the basis contrast;
Except that BAS and Cont group, all the other groups are continuous 3d filling stomach vitamin D during experiment the 53rd～55d all
3Parenteral solution (1 * 10
6Ukg
-1D
-1);
(1) Cont group: press table 1.1 medication continuous irrigation stomach during the experiment beginning, irritate stomach time and V.H group synchronously;
(2) V.H group: press table 1.1 medication continuous irrigation stomach 38d during the experiment beginning.Behind the conventional raising 2w, vitamin D
3Parenteral solution (1 * 10
6Ukg
-1D
-1) continuous irrigation stomach 3d.Press table 1.1 medication filling stomach after observing 5d, continuously 4w;
(3) V.H+XZK group: press table 1.1 medication continuous irrigation stomach during the experiment beginning, irritate stomach time and V.H group synchronously;
(4) V.H+RK group: press table 1.1 medication continuous irrigation stomach during the experiment beginning, irritate stomach time and V.H group synchronously;
(5) V.H+Epi. group: press table 1.1 medication continuous irrigation stomach during the experiment beginning, irritate stomach time and V.H group synchronously;
(6) V.H+R.E group: press table 1.1 medication continuous irrigation stomach during the experiment beginning, irritate stomach time and V.H group synchronously.
1.3 the preparation of medicine and preparation
1.3.1 blood fat recovering particles suspension
Commercially available blood fat health drug particles is mixed with the required blood fat recovering particles suspension of experiment, per 1 commercially available blood fat recovery capsule medicine (0.3g/ grain) adding distil water 14ml.4 ℃ of refrigerators are preserved subsequent use.Use in one week.
1.3.2 Monascus anka Nakazawa et sato filament extract mixture suspension
The Monascus anka Nakazawa et sato filament extract pulverizing of getting the present invention's preparation is fine powder, and every 2g red colouring agent for food, also used as a Chinese medicine fine powder adds 7ml distilled water and is mixed with the required Monascus anka Nakazawa et sato filament extract mixture suspension of experiment.4 ℃ of refrigerator preservations are subsequent use, use in the week.
1.3.3 barrenwort alcohol extract
The preparation of barrenwort alcohol extract: barrenwort, pulverize, add 15 times of 70% alcohol immersion 24 hours, heating and refluxing extraction 2 times is filtered, and keeps filtrating.Filter residue adds 40% alcohol immersion 24 hours again, and heating and refluxing extraction 1 time is filtered, and keeps filtrating, merges filtrating 3 times, reclaims ethanol, is concentrated into 100%.(the concentration of 1g crude drug/ml).4 ℃ of refrigerators are preserved subsequent use.
1.3.4 Monascus anka Nakazawa et sato filament extract and barrenwort suspension
Barrenwort alcohol extract and Monascus anka Nakazawa et sato filament extract mixture suspension are processed the back equal-volume and are mixed, and 4 ℃ of refrigerators are preserved subsequent use.
1.4 experimental technique
1.4.1 fluorescence labeling method and animal are drawn materials in the rat body
All animals weigh in weekly once; Stomach dosage is irritated in adjustment according to body weight.Finish preceding 14d, each row neck hypodermic injection quadracycline (37.5mgkg of 13d in experiment
-1D
-1), as the 1st fluorescence labeling, finish preceding 4d, each row neck hypodermic injection calcein (10mgkg of 3d in experiment
-1D
-1) as the 2nd fluorescence labeling, forming yellow and green two fluorized markings at bone surface, be 10d twice fluorescence labeling blanking time.When experiment finishes, with 3% yellow Jackets (4mgkg
-1) the intraperitoneal injection of anesthesia rat, heart extracting blood is peeled off the total length artery along aorta petal to common iliac artery bifurcation after putting to death rat, gets aorta pectoralis, is used for morphological examination; Take off bilateral femur, left side shin bone rapidly, remove muscle, carry out bone morphometry index, bone biomechanical parameter, ossein and content of mineral substances as follows and measure.
1.4.2 the making of rat chest aorta section
Get the about 1cm of aorta pectoralis hypomere immediately with the fixing 24h of 10% neutral formalin buffer solution, carry out the gradient dehydration according to following order, transparent; Embedding: 95% ethanol (1.5h), 95% ethanol (1.5h), 100% ethanol (0.5h); 100% ethanol (0.5h); Xylenes (15min), waxdip (3h), embedding.The finishing wax stone, subsequent use.Section: paraffin mass is fixed on the slicer, cuts the section of 4 μ m thickness, and the exhibition sheet is on the slide that glycerine albumen was smeared, and desire row HE dyeing and von Kossa dye.
1) the HE dyeing course is following:
Xylenes I (10min), xylenes II (10min), 100% ethanol (3min), 100% ethanol (3min), 95% ethanol (2min); 80% ethanol (2min), 70% ethanol (2min), distillation washing, haematoxylin (10min), distillation washing; Acidic alcohol (3s), flowing water is washed (30min～2h), distill washing, 70% ethanol (2min), 80% ethanol (2min) immediately; Yihong (5min), the distillation washing, air duct dries up, and xylenes soaks into, mounting.
2) von Kossa dyeing course is following:
Xylenes I (10min), xylenes II (10min), 100% ethanol (3min), 100% ethanol (3min), 95% ethanol (2min), 80% ethanol (2min), 70% ethanol (2min), distillation washing, 5%AgNO
3The irradiation down of the aqueous solution (10min), uviol lamp (1～2h), the distillation washing, 1% dimethyl diaminophenazine chloride (3min), the distillation washing, air duct dries up, and xylenes soaks into, mounting.
1.4.3 the assay method of serum biochemistry index
Test 29d, 39d eye socket and get blood dynamic monitoring serum index and confirm blood lipid level, fasting 12h weighs in after the last administration; After 3% yellow Jackets lumbar injection (4mg/kg) anesthesia, through heart extracting blood 8～10ml, leave standstill 4h after; The centrifugal 10min of 3500rpm/min; Separate upper serum, packing serum is respectively applied for detection: the content of serum total cholesterol (TC), HDL-C (HDL-c), LDL-C (LDL-c), triglycerides (TG), calcium (Ca), phosphorus (P); Take colorimetric method to detect, concrete operations are undertaken by the kit specification.
1.4.4 undecalcified bone tissue section preparation
Take out the right shin bone of rat rapidly after getting blood, muscle and connective tissue are adhered in separation, Ex-all, open pulp cavity with sawing at near-end at a slow speed as sagittal, put into 10% neutral formalin buffer solution and fix, and are no more than 24h.By following method dehydration, plastic embedding is done in degreasing then.Downcut undecalcified 4 μ m and 9 μ m section respectively with sclerous tissues's slicer, the cma staining mounting is used in 4 μ m section, carries out bone meterological static parameter (static paramet-ers) and measures.The 9 μ m section mounting that do not dye carries out bone meterological dynamic parameter (dynamicpara-meters) as Fluirescence observation and measures.The preparation detailed method:
(1) elution process of monomer (methyl methacrylate) before the embedding
1. the methyl methacrylate of 1500ml is poured in the separatory funnel (3L).2. add 5%NaOH solution 500ml, fully shake up, leave standstill, treat the solution layering after, emit lower floor's solution, discard.3. repetitive operation is 1. three times, and three times total amount and amount of monomer be (wash-out retarding agent) quite.4. add the water repetitive operation 2. three times (wash-out NaOH) of equivalent.5. the monomer of above-mentioned processing is put into anhydrous CaCl
2Dehydration is 2 times (1000ml/500g).Filter paper filters, and collects filtrating.6.-20 ℃ preservation is subsequent use, takes out in second day and to see that to have or not ice crystal floating, does not have, show anhydrous can be subsequent use, have, then need dehydration again.
(2) the preparation process before the embedding of shin bone epimere
1. expose ossis: the shin bone in the fixer is cut with low speed, expose ossis;
2. fix and dehydration: respectively through 70% ethanol 1d, 70% ethanol 1d, 95% ethanol 2d, 100% ethanol 1d, five dehydrations of 100% ethanol 1d, the transparent 1d of xylenes.
3. infiltration: (methyl methacrylate 90ml, dibutyl phthalate 10ml stir 3h on the magnetic stirring apparatus successively to use immersion liquid I.), (the stupid formyl chloride-1 .0g of peroxidating stirs 4.5h to immersion liquid II on the magnetic stirring apparatus for methyl methacrylate 90ml, dibutyl phthalate 10ml.), (benzoyl peroxide 2.5g stirs 6h to immersion liquid III on the magnetic stirring apparatus for methyl methacrylate 90ml, dibutyl phthalate 10ml.) soak into 3d respectively.
(3) embedding
Pour into to be equipped with the immersion liquid III (fresh configuration on the same day) of fresh configuration and soak into good bone piece volume and be about in the bottle of 15ml; The about 1.5cm of the embedding medium of pouring into is high; Build bottle cap, and on bottle cap, insert a syringe needle (note: syringe needle can not submerge in the immersion liquid), ambient temperature overnight.Inserted the embedding bottle in second day polymerization in 25 ℃ the constant temperature roaster; Beginning is every raises 1 ℃ of oven temperature at a distance from 1.5h, the most to a high-profile to 39 ℃, during adjustment more backward blanking time long more; Form until embedded block; As producing bubble in the embedding process, available 1ml syringe sucks embedding liquid and squeezes in the bubble, and bubble is released.The shin bone epimere needs that it is cut face and pastes the central authorities' embedding of the bottle end, pours the high embedding medium of about 3.5cm into and gets final product.
(4) preparation of slide
1. clean: in sour cylinder, soak 48h to slide, take out, wash 3 times with running water earlier, the back is washed 2 times with single water that steams, and distilled water flushing 4 times is dried, 75% alcohol disinfecting, dry for standby (perhaps ultraviolet ray irradiation 2h is subsequent use).
2. gluing: add the 9g gelatin in the 1000ml distilled water, heating makes the gelatin dissolving, and other 4% clear and bright potassium chromium sulfate solution of preparation is joined in the gelatin solution of boiling; Constantly mix liquid; After treating the solute dissolving, stop heating, treat that solution is cooled to can use about 50 ℃; The wave carrier piece of hand basket of will packing into soaked 2 minutes in 50 ℃ glue, took out and put into cool sheet plate, airing subsequent use (visual actual conditions adjustment gluing number of times).
(5) section
1. with the gypsum sander embedded block is polished into suitable size, and exposes tangent plane, be polished to correct position to tangent plane.2. the good embedded block of will polishing is fixed on the slicer, drips 40% alcohol and treats that in embedded block tangent plane is lubricated once, and the right hand slowly downcuts; Left hand is dialled down section gently with a banister brush, notes not doing the mashed sheet that downcuts, and transferring to sheet to drip with hairbrush then has on the slide of one 70% alcohol; Drip 70% alcohol again; Slowly flatten section then, cover a thin plastic sheet, and unnecessary alcohol is blotted with filter paper.One group of sheet that 3. will cut is used clamp, puts into about the roasting 24～48h of air blast baking box about 40 ℃, and taking-up ties up subsequent use with rubber band.4. sample is cut the sheet 4 μ m (thin slice) and the 9 μ m (sheet) of two kinds of thickness, and thin slice is used for Masson-Goldner Trichrome dyeing or cma staining, the transparent mounting of the direct xylenes of sheet.
(6) osteocomma dyeing
The thin slice of 4 μ m often adopts cma staining or Masson-Goldner Trichrome dyeing, and after latter's dyeing, sclerotin is green, and marrow is red, is used to observe the surperficial osteoclast of bone trabecula and measures its girth.Be Masson-Goldner Trichrome dyeing course below.
1. the preparation of working solution
A.Weigert he matoxylin working solution: solution A contain Mematoxylin (haematoxylin smart) 1g add filter among the 95% alcohol 100ml subsequent use; Solution B contains 29% iron chloride 4ml, watery hydrochloric acid 1ml (dilution in 1: 4 of the hydrochloric acid adding distil water with 40%), and adding distil water is to 100ml then.The solution A and the solution B mixing of equal portions are promptly got working solution.
B.Poncean Fuchsin Stock working solution: obtain solution A contains Poncean (Ponceaux) 1g adding distil water 100ml, and solution B contains Acid Fuchsin (magenta) 1g adding distil water 100ml; The solution B of 3 parts solution A and 1 part mixed being made into stoste, then stoste is become working solution with 0.2% acetic acid by 5: 1 dilution proportion.
C.Phosphotungstic acid-Orange G working solution: Orange G 2g and Phosphotungstic acid 4g are added in the 100ml distilled water, use preceding filtration.
D.Light green working solution: Light green (bright green) 0.2g and acetic acid 0.2ml are added in the 100ml distilled water, use preceding filtration
2. dyeing course
1. take off the section of moulding and be placed on the dyeing of Weigerts hematoxylin working solution, 25min, distilled water rinsing; 2. running water rinsing, 10min; 3. distilled water rinsing; 4. PoNSean-Fuchsin working solution dyeing, 13-15min; 5. 1% acetic acid rinsing is 2 times, each 1min; 6. the Orange G liquid of fresh filtration dyeing 7min; 7. the Light green of fresh filtration dyeing 15-20min; 8. distillation washing, air duct dries up; 9. xylenes cleans 2 times, each 2min, mounting.
1.4.5 cortex bone undecalcified bone abrasive disc preparation method
Abrasive disc is that middle tibia is sawn into the thick osteocomma of 200 μ m with veneer sawing machine earlier, is separated into effective osteocomma fully, the error [47] that this is caused in the time of can reducing section as far as possible from the junctura tibiofibularis proximal end with dried lower end of shin bone and fibula during the saw bone.Saw three altogether, the tibiofibula junction is first, then down is followed successively by second and the 3rd.With second thin slice of wearing into 30 μ m-50 μ m, dry up the transparent back of xylenes resinene mounting again with air duct with coarse glass.Measure the parameters of cortex bone.
1.4.6 bone tissue morphometry index determining
(1) shin bone epimere measurement category
From the growth plate 2.0mm that down draws,, down draw 3.0mm as shin bone epimere measurement category from 2.0mm again to avoid primary sponge.
(2) dynamic and static parameter measurement and calculating
All parameters of this experiment adopt the name of general international standard bone tissue norphometry technics, and the Chinese and English title, symbol and the unit that directly measure parameter with semi-automatic image digitizer are like shown in the table 1.2.But these parameters can't directly be used to analyze bone amount, bone structure, bone formation and bone absorption etc., and the calculation of parameter that directly record through international formula just can be used for analyzing.Chinese and English title, symbol and the computing formula of calculating parameter is like shown in table 1.2 and the table 1.3.
Table 1.2 cancellous bone measurement parameter
1.3 cancellous bone calculating parameter and computing formula
(3) meaning of parameter
1. static parameter: be used for estimating the medical treatment effect, describe the bone amount what with bone trabecular morphosis.
Bone trabecula area percent (%Tb.Ar): phalanges girder area accounts for the percentage of bone tissue area, and the bone amount what reflect.Calculate from mathematical formulae, it equal bone trabecula width and quantity product 1/10, that is to say, the bone amount what by width and the two common decision of quantity.
Bone trabecula thickness (Tb.Th): be used to describe the trabecular bone structure form, explain the bone quantitative changeization.Its variation can influence the bone amount, and under the certain condition of quantity, width is wide more, and the bone amount is many more.
Bone trabecula quantity (Tb.N): be used to describe the trabecular bone structure form, explain the bone quantitative changeization.Its variation can influence the bone amount, and under the certain condition of width, quantity is many more, and the bone amount is many more.
Bone trabecula separating degree (Tb.Sp): the average distance between the phalanges girder is used for describing the trabecular bone structure form.Separating degree is big more, and distance is just big more between the bone trabecula, and bone is just loose more.
2. dynamic parameter: dynamic parameter comprises that bone forms parameter and bone absorption parameter, through dynamic parameter, can understand the amount and the speed of bone surface mineralising, and explains the reason of static parameter variation.
Perimyelis fluorescent perimeter percentage (%L.Pm): reflect that the bone girth that carries out mineralising accounts for the percentage of bone surface overall circumference, also reflects osteoblastic quantity.
Perimyelis mineralising deposition (MAR): the width that refers to mineralising every day.The speed of reflection bone mineralising is represented osteoblastic activity.
Bone formation rate (BFR/BS): the product of %L.Pm and MAR, the active degree of representing bone surface to form.
Bone formation rate (BFR/BV): the annual new bone trabecula area that forms accounts for the percentage of the bone trabecula gross area, represents the active degree that bone forms and bone is changed.This more parameters is high, and the conversion of expression bone is also high more.It is the osteoplastic important indicator of reflection.
Bone formation rate (BFR/TV): the annual new bone trabecula area that forms accounts for the percentage of bone tissue area, represents the new total amount that forms bone in bone for year girder surface.This parameter and BFR/BS, BFR/BV are different, and it receives two factor affecting, and the one, what of bone amount, the bone amount is more, and just to provide more new bones to form required bone trabecula surperficial; The 2nd, bone forms active degree, and bone forms active more, and the new bone that on unit length, forms is just many more.Though so bone formation sometimes is very active, because the bone amount very little, this parameter also can descend.Can draw from the formula reasoning, BFR/TV equals 1/100 of %Tb.Ar and BFR/BV product.
1.4.7 the assay method of femur weight
Get the left side femur, muscle and the soft tissue on the bone adhered in Ex-all, claims to examine 72h to constant weight in 80 ℃ of baking box bakings after its weight in wet base with the AE240 electronic balance, and title bone this moment is heavy can to draw the femur dry weight.
1.4.8 bone hydroxyproline (Hyp) and bone calcium (Ca), bone phosphorus (P), bone magnesium (Mg) assay method
After measuring left side femur weight; Place the 10ml ampoule bottle to add 6mol/LHCl 5ml every part of bone specimen then; Alcohol blast burner is burnt the sealing of ampoule bottleneck and is placed in the baking box; Take out behind 108 ℃ of constant temperature digestion 24h and filter, get filtrating, then filtrating is divided into two parts; A with 1: 400 dilution back concentration with elements such as ICP (inductance coupled plasma direct-reading spectrometer) mensuration bone Ca, bone P, bone Mg, calculate the content of elements such as bone Ca, bone P, bone Mg according to standard items concentration and sample extension rate.Regulate pH value to 6.8～7.0 backs after another part filtrating is diluted 10 times according to the absorbance of hydroxyproline determination kit method, calculate the content of hydroxyproline according to formula with hydroxyproline in the ELIASA working sample.
1.4.9 bone density detection method
Get rat right side femur after putting to death rat, preserve moisture, be stored in-20 ℃ with physiological saline wet gauze parcel.During test, the femur normal temperature unfreezing of preserving is wet again with physiological saline, and Nuclear Medicine Dept of overseas Chinese hospital measures femoral bmd with PRODIGY type borne densitometers (precision of measurement is less than 1%) and draws bone density value (BMD) in Guangzhou then.Experiment is this time chosen femur entirely as the BMD measurement point.Rat femur after measuring is still sealed with gauze, with being put in-20 ℃ of preservations, bone biomechanical index to be measured behind the tinfoil parcel.
1.4.10 bone biomechanical assay method
The three-point bending experiment
During test, with the femur normal temperature unfreezing of-20 ℃ of preservations, physiological saline is wet again.Analyze the biomechanical property (assisting to accomplish) of right side femur (three-point bending experiment) by biomethanics National Key Laboratory of Nanfang Medical Univ with 858Mini Bionix type material test macro.
(1) experimental technique
Rat right side femur is placed on the MTS testing machine, and with the pressure head of 1mm diameter, loading velocity is 0.01mm/s, and span (L) is 15mm.Instrument writes down the load (F) and oar degree (d) changing value of each moment point automatically, drafting load-and around the line of writing music, and obtain elastic load, maximum load, breaking load, elastic modelling quantity and stiffness coefficient etc.
(2) measurement of parameter and calculating
Biomechanical property CALCULATION OF PARAMETERS formula in table 1.4 three-point bending method
(3) parameter meaning
Bone strength: the structural mechanics characteristic of bone, relevant with the material behavior of bone, also receive geometrical property (shape, the size) influence of bone.Comprise: elastic load: the greatest force that bone can bear in elastic range; Maximum load: the greatest force of being born before the bone fracture; Breaking load: the external force of being born during the bone fracture; Rigidity: be called external hardness again, be elastic deformation area's slope, refer to resist the bone specimen deformation ability, receive the influence of bone size, mainly reflect the material behavior of bone tissue; Elastic modelling quantity: claim inherent hardness again.The ratio of elastic limit internal stress and corresponding line strain is represented the ability of the resistance to deformation of material.
Compression experiment
(1) experimental technique
Get the rat spine fifth lumbar vertebra, remove neural spine, transverse process and articular cartilage and (notice that protection centrum crust mass shell avoids infringement, because it has material impact to the vertebra mechanical property.Centrum is made into the high cylinder of about 6mm with parallel end, and the sample of manufacturing has like this been removed chondrosin long slab and elementary cancellous bone, only is made up of the trabecular bone at center and the cortex shell of periphery.Measure its volume and height with buoyancy method, and calculate cross-sectional area (J).Last lumbar vertebrae is placed on the MTS testing machine, with the pressure head of 1mm diameter, is pressed in the cancellous bone of cone record load-deflection curve, performance indications such as acquisition maximum compression load, maximum strain, elastic modelling quantity, bone hardness with the speed of 0.01mm/s.
The measurement of parameter and calculating
Biomechanical property CALCULATION OF PARAMETERS formula in table 1.5 compression method
(3) parameter meaning
Maximum compression load: the greatest force that test specimen can bear is to bone trabecular sclerotin, the concentrated expression of structural continuity, cortical thickness, cross-sectional area and quality of materials; Elastic modelling quantity: the ratio of elastic limit internal stress and corresponding line strain, represent the ability of the resistance to deformation of material; Bone strength: the greatest force that unit bone amount can bear, the quality of materials of reflection bone; Bone hardness: the elastic modelling quantity of unit bone amount, the inherent non-deformability of reflection bone material, irrelevant with the bone amount; Absorption maximum energy: make the energy of the required absorption of sample compression fracture, represent the toughness of bone material, receive bone mineral content content, the influence that bone matrix compositions such as collagenous fibres trend change.
1.5 statistical analysis
Expression that experimental data is all used
; Adopt SPSS16.0 software; Relatively adopt one-way analysis of variance (ANOVA) between the group of normal distribution data, the The data LSD check that variance is neat; The data of heterogeneity of variance then adopt Tamhane ' s T2 check, and with P＜0.05, P＜0.01 expression has statistical significance.
2 results
2.1 the general situation of experimental rat
Experimental rat generally in order; Each organizes activities in rats, it is normal to drink water, ingest, and all super physiologic dosage cholecalciferol parenteral solutions that gives are irritated the stomach group, and in various degree One's spirits are drooping depressed appears in rat; Be off one's feed; Do not like, indivedual rat eye sockets are hemorrhage or dead, but individual total coincidence statistics requirement in the group.
2.2 the Monascus anka Nakazawa et sato filament extract is to the influence of rat body weight
The Monascus anka Nakazawa et sato filament extract is seen table 2.1 to the influence of rat body weight:
Table 2.1 Monascus anka Nakazawa et sato filament extract is to the influence (
g) of rat physique amount
Shown in table 2.1 result, except that control group, all the other groups are after the experiment beginning, and body weight strengthens than control group amplification, surpassed control group during to 12d, 24d, and 52d begins the vitamin D that continuous 3d gives respectively to organize rats gavaged super physiologic dosage in experiment
3Behind the parenteral solution, rat body weight sharply descends, and shows as poor appetite, and the is One's spirits are drooping state of Denging; This moment, the control group body weight was higher than all the other group rats, and behind the observation 5d, each is organized the rat state and takes a turn for the better, and continues to irritate the high lipoprotein emulsion of stomach; The body weight bottom out behind 74d, all surpasses control group basically.But not statistically significant between each group of rat body weight.
2.3 the Monascus anka Nakazawa et sato filament extract is to the influence of rat blood serum lipid
The Monascus anka Nakazawa et sato filament extract is seen table 2.2 and table 2.3 to the influence of rat blood serum lipid:
Table 2.2 Monascus anka Nakazawa et sato filament extract is to the influence
of rat blood serum lipid
Annotate:
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+XZK group P＜0.05P＜0.01.
Table 2.3 Monascus anka Nakazawa et sato filament extract is to the influence
of rat blood serum lipid
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+XZK group P＜0.05P＜0.01
Shown in table 2.2 and table 2.3,
(1) model+blood fat health group and model group compare, and serum total cholesterol, serum low-density LP cholesterol level reduce about 43.6% (P＜0.01), 39.4% (P＜0.01) respectively; The ratio of the ratio of serum high-density LP and serum total cholesterol, serum high-density LP and serum low-density LP cholesterol raise respectively about 77.8% (P＜0.01), 237.5% (P＜0.01).
(2) model+Monascus anka Nakazawa et sato filament extract group and model group are relatively; Serum total cholesterol, serum low-density LP cholesterol level reduce about 49.4% (P＜0.01), 78.0% (P＜0.01) respectively, the ratio of the ratio of triglycerides, serum high-density LP cholesterol level, serum high-density LP and serum total cholesterol, serum high-density LP and serum low-density LP cholesterol raise respectively about 9.4% (P＜0.05), 34.0% (P＜0.01), 177.8% (P＜0.01), 1012.5% (P＜0.01).
(3) model+Monascus anka Nakazawa et sato filament extract group is compared with model+blood fat health group; The serum low-density LP cholesterol level reduces about 63.8% (P＜0.01), the ratio of serum high-density LP cholesterol level, serum high-density LP and serum total cholesterol raise respectively about 50.0% (P＜0.05), 56.3% (P＜0.05).
(4) model+barrenwort group and model group compare, and serum total cholesterol, serum low-density LP cholesterol level reduce about 50.4% (P＜0.01), 81.8% (P＜0.01) respectively; The ratio of the ratio of triglycerides, serum high-density LP cholesterol level, serum high-density LP and serum total cholesterol, serum high-density LP and serum low-density LP cholesterol raise respectively about 46.9% (P＜0.01), 27.7% (P＜0.01), 177.8% (P＜0.01), 1575.0% (P＜0.01).
(5) model+barrenwort group is compared with model+blood fat health group; The serum low-density LP cholesterol level reduces about 70.0% (P＜0.01), the ratio of serum high-density LP cholesterol level, serum high-density LP and serum total cholesterol raise respectively about 42.9% (P＜0.05), 56.3% (P＜0.05).
(6) model+Monascus anka Nakazawa et sato filament extract+barrenwort group and model group compare, and serum total cholesterol, serum low-density LP cholesterol level reduce about 51.6% (P＜0.01), 84.1% (P＜0.01) respectively; The ratio of the ratio of triglycerides, serum high-density LP cholesterol level, serum high-density LP and serum total cholesterol, serum high-density LP and serum low-density LP cholesterol raise respectively about 28.1% (P＜0.05), 38.3% (P＜0.01), 188.9% (P＜0.01), 1862.5% (P＜0.01).
(7) model+Monascus anka Nakazawa et sato filament extract+barrenwort group is compared with model+blood fat health group; The serum low-density LP cholesterol level reduces about 73.8% (P＜0.05), the ratio of serum high-density LP cholesterol level, serum high-density LP and serum total cholesterol raise respectively about 54.8% (P＜0.01), 62.5% (P＜0.01).
2.4 the Monascus anka Nakazawa et sato filament extract is to the influence of rat blood serum calcium, phosphorus concentration
The Monascus anka Nakazawa et sato filament extract is seen table 2.4 to the influence of rat blood serum calcium, phosphorus concentration:
Table 2.4 Monascus anka Nakazawa et sato filament extract is to rat blood serum calcium, phosphorus concentration variation
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+XZK group P＜0.05P＜0.01
Shown in table 2.4 result,
(1) model+blood fat health group and model group relatively, serum calcium levels there was no significant difference, serum paraoxonase content about 7.2% (P＜0.01) that raises.
(2) model+Monascus anka Nakazawa et sato filament extract group and model group compare, and serum calcium levels has reduced by 22.3% (P＜0.05), and serum paraoxonase content reduces about 9.6% (P＜0.01).
(3) model+Monascus anka Nakazawa et sato filament extract group is compared with blood fat health group, and serum paraoxonase content reduces by 15.6% (P＜0.01).
(4) all the other groups and model group, model+blood fat health group relatively, serum calcium, phosphorus content there are no significant difference.
2.5 the Monascus anka Nakazawa et sato filament extract is to the influence of rat artery Pathomorphology
The Monascus anka Nakazawa et sato filament extract is following to the morphological change of rat chest aorta vascular wall influence:
Control group: vascular wall is complete, smooth muscle marshalling and closely;
Model group: membrane portions is destroyed the cell arrangement disorder in the aorta pectoralis; The obviously atrophy of middle membrane portions zone, the elastic fibrous layer structure is unclear, arranges to loosen, the ductility reduction, proliferation of fibrous tissue companion's sheet or punctate clacification are typical calcified plaque;
Model+blood fat health group, the inside and outside wall profile is still complete, and elastomer shrinks or fracture.
Model+Monascus anka Nakazawa et sato filament extract group, adventitia is loose, and elastic fibers layer clear in structure is complete, and well arranged but ductility reduces;
Model+barrenwort group: inner membrance and outer membrane structure are unclear, the cell arrangement disorder, and middle membrane portions zone elastic fibrous layer and muscle fibre layer structure are unclear, and ductility reduces, proliferation of fibrous tissue companion punctate clacification;
Model+Monascus anka Nakazawa et sato filament extract+barrenwort group: the inner membrance clear in structure, adventitia is damaged, middle film elastic fibers layer clear in structure, the part fracture, ductility reduces, and has the sheet calcification to occur;
2.6 the Monascus anka Nakazawa et sato filament extract is to the influence of rat tibia bone morphometry static parameter
The Monascus anka Nakazawa et sato filament extract is seen table 2.5 to the influence of rat tibia bone morphometry static parameter:
Table 2.5 Monascus anka Nakazawa et sato filament extract is to rat tibia morphology proximal meterological static parameter
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+Epi. group P＜0.05P＜0.01
Shown in table 2.5 result:
(1) model+barrenwort group and model group are relatively; Bone trabecula area percent (%Tb.Ar), bone trabecula number (Tb.N) increase about 53.8% (P＜0.01), 36.9% (P＜0.05) respectively; Bone trabecula separating degree (Tb.Sp) has reduced by 38.8% (P＜0.01); Bone trabecula thickness (Tb.Th) has increase trend, but not obvious;
(2) model+Monascus anka Nakazawa et sato filament extract group and model group compare, and bone trabecula area percent, bone trabecula thickness increase about 30.1% (P＜0.01), 19.2% (P＜0.05) respectively;
(3) model+Monascus anka Nakazawa et sato filament extract group is compared with model+barrenwort group, and bone trabecula area percent, bone trabecula number have reduced by 15.42% (P＜0.05), 19.08% (P＜0.05) respectively, bone trabecula separating degree 34.03% (P＜0.05) that raise;
(4) model+Monascus anka Nakazawa et sato filament extract+barrenwort group and model group compare, and bone trabecula area percent, bone trabecula thickness increase about 37.8% (P＜0.01), 12.7% (P＜0.05) respectively, and the bone trabecula separating degree has reduced by 25.3% (P＜0.05).
2.7 the Monascus anka Nakazawa et sato filament extract is to the influence of rat tibia bone morphometry dynamic parameter
The Monascus anka Nakazawa et sato filament extract causes that the variation of rat tibia bone morphometry dynamic parameter sees table 2.6:
Table 2.6 proximal tibia morphometry dynamic parameter
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+Epi. group P＜0.05P＜0.01
Shown in table 2.6:
(1) model+barrenwort group and model group compare, and perimyelis fluorescent perimeter percentage (%L.Pm), perimyelis mineralising deposition (MAR), bone formation rate (BFR/TV), bone formation rate (BFR/BS) increase about 51.8% (P＜0.01), 83.9% (P＜0.05), 71.0% (P＜0.05), 106.7% (P＜0.05) respectively.
(2) model+Monascus anka Nakazawa et sato filament extract group and model group compare, and perimyelis fluorescent perimeter percentage, bone formation rate (BFR/TV), bone formation rate (BFR/BS) increase about 48.6% (P＜0.01), 94.3% (P＜0.05), 70.4% (P＜0.05) respectively.
(3) model+Monascus anka Nakazawa et sato filament extract+barrenwort group and model group compare, and perimyelis fluorescent perimeter percentage, perimyelis mineralising deposition increase about 37.6% (P＜0.05) respectively, 61.3% (P＜0.05), 71.0% (P＜0.05).
(4) model+Monascus anka Nakazawa et sato filament extract+barrenwort group is compared with model+barrenwort group, and bone formation rate (BFR/TV) has reduced by 39.67% (P＜0.05).
2.8 the Monascus anka Nakazawa et sato filament extract is to the influence of rat femur bone mineral and ossein content
The Monascus anka Nakazawa et sato filament extract is seen table 2.7 to the variation of rat femur bone mineral and ossein content:
Table 2.7 Monascus anka Nakazawa et sato filament extract is to the influence
of rat femur bone mineral and ossein content
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Carefully compare with V.H P＜0.01; Compare with the V.H+Epi. group P＜0.05P＜0.01
Shown in table 2.7 result,
(1) model+barrenwort group and model group compare, and calcium content of bone increases about 11.3% (P＜0.05).
(2) model+functional red yeast rice preparation group and model group compare, and bone calcium, bone content of magnesium increase about 4.9% (P＜0.05), 6.3% (P＜0.05) respectively.
(3) model+functional red yeast rice preparation+barrenwort group and model group compare, and the bone content of magnesium increases about 7.2% (P＜0.05).
2.9 the Monascus anka Nakazawa et sato filament extract is to the influence of rat femur bone density
The Monascus anka Nakazawa et sato filament extract is seen table 2.8 to the rat femur changes of bone mineral density:
Table 2.8 Monascus anka Nakazawa et sato filament extract is to the influence (
n=10) of rat femur density
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+Epi. group P＜0.05P＜0.01
Shown in table 2.8 result,
(1) model+barrenwort group and model group relatively, lumbar spine bmd 22.7% (P＜0.01) that raise
(2) model+functional red yeast rice preparation group is compared with model+barrenwort group, and lumbar spine bmd has reduced by 11.11% (P＜0.05) and compared with model group, there was no significant difference between each group of femoral bmd.
2.10 the Monascus anka Nakazawa et sato filament extract is to the influence of rat biomechanical parameter
The Monascus anka Nakazawa et sato filament extract is seen table 2.9 and table 2.10 to the variation of rat biomechanical parameter:
Table 2.9 Monascus anka Nakazawa et sato filament extract is to the influence
of rat biomechanical parameter
*P＜0.05
*Compare with the Cont group P＜0.01;
▲P＜0.05
▲ ▲Compare with the V.H group P＜0.01; Compare with the V.H+Epi. group P＜0.05P＜0.01
Table 2.10 Monascus anka Nakazawa et sato filament extract is to the influence
of rat biomechanical parameter
*P＜0.05
*Compare with the Cont group P＜0.01; ▲ P＜0.05 ▲ ▲ P＜0.01 and the comparison of V.H group; Compare with the V.H+Epi. group P＜0.05P＜0.01
Shown in table 2.9 and table 2.10,
(1) model+barrenwort group and model group compare, and elastic load, maximum load, elastic modelling quantity have increased by 14.9% (P＜0.05), 12.8% (P＜0.05), 37.5% (P＜0.05) respectively.
(2) model+Monascus anka Nakazawa et sato filament extract group and model group compare, and maximum load, breaking load, stiffness coefficient have increased by 13.2% (P＜0.05), 19.4% (P＜0.05), 31.9% (P＜0.05) respectively.
3 discuss
3.1 the Monascus anka Nakazawa et sato filament extract is to the discussion of high fat+atherosis preventive and therapeutic effect of cholecalciferol rat artery
By experimental result: the high fat of Monascus anka Nakazawa et sato filament extract for treating+cholecalciferol rat; Experiment finishes the back to be compared with the model group rat, and serum TC, LDL-c obviously descend, HDL-c, HDL-c/TC, HDL-c/LDL-c obviously raise (P＜0.05); Blood vessel inside and outside wall profile is complete basically; Elastomer is loose, the minority fracture, and atheromatous plaque forms less; With blood fat health group ratio, the functional red yeast rice preparation reduces LDL-c, and rising HDL-c effect is more obvious, and the interior calcified plaque of blood vessel is littler, still less.
Thus, we think that the Monascus anka Nakazawa et sato filament extract can effectively reduce high fat+cholecalciferol rat fat, prevents and treats atherosclerotic.
The Monascus anka Nakazawa et sato filament extract is identified through other teachers of this seminar and is not wherein contained Lovastatin, possibly contain multiple unrighted acid, cholesterine, ubiquinone
10, multiple hypotensive, blood sugar reducing component such as various trace elements, glycosaminoglycan, GABA, dietary fiber.Experimental result shows that the Monascus anka Nakazawa et sato filament extract can significantly reduce TC, LDL-c, rising HDL-c, HDL-c/TC, HDL-c/LDL-c, and the protection blood vessel reduces the formation of atherosclerotic plaque.Infer its study of anti-atherogenic effect mechanism maybe for: 1) suppress the HMG-CoA reductase.This Monascus anka Nakazawa et sato filament extract possibly contain the Monacolins class material except that Lovastatin, is hydroxyl first glutaryl coenzyme A (HMG-CoA) reductase inhibitor active component, has limited the synthetic speed of cholesterol; 2) through inhibition to cholesterol biosynthesis rate-limiting enzyme activity; Thereby reduce the concentration of liver cell inner cholesterol; Regulate through negative-feedback; The ldl receptor expression number and the activity that have suppressed surface of hepatocytes, result make LDL and precursor intermediated-density lipoprotein thereof, and (Intermediate density lipoprotein IDL) generates minimizing; The change VLDL (Very low-density lipoprotein, composition VLDL), and make it generate minimizing, from serum, removing increases; The HDL level that also raises is transferred fat and effect for reducing fat thereby reach; 3) when transferring fat, possibly reduce the inflammatory factor level in the patients with coronary heart disease body, help preventing atherosclerotic generation; 4) booster action of Co-Q10.Red colouring agent for food, also used as a Chinese medicine makes the synthetic minimizing of cholesterol; Then supply with the also corresponding minimizing of cholesterol of cell membrane; The permeability and the unstability that cause cell membrane increase, and cell function is unusual, and having increased the weight of in blood circulation mainly by LDL-c is the shortage of the Co-Q10 that transports of carrier protein.Contain Co-Q10 in this functional red yeast rice preparation, have anti-oxidant, remove free radical, repair injured neuron, improve important effects such as immunity of organisms
[81], also stoped atherosclerotic plaque formation to a certain extent.
In view of the above; We think that the Monascus anka Nakazawa et sato filament extract has the ability that potential control atherosclerotic lesion takes place, develops; Can reduce the incidence of disease and the death rate of cardiovascular and cerebrovascular disease; The effect of Monascus anka Nakazawa et sato filament extract possibly be many-sided, and its antiatherogenic mechanism also needs further to inquire into, for the generation that prevents and treats the atherosclerotic angiocardiopathy clinically provides new theoretical foundation.
3.2 the Monascus anka Nakazawa et sato filament extract is to high fat+vitamin D
3The osteoporosis rat preventive and therapeutic effect
Can know the high fat+vitamin D of Monascus anka Nakazawa et sato filament extract gastric infusion by this result of experiment
3Rat, experiment is compared with the model group rat after finishing, and the bone amount increases, and is especially obvious to the effect of cancellous bone, improved bone trabecular microstructure through increasing bone trabecula thickness, and lumbar spine bmd increases, and the biomechanical property of bone makes moderate progress.Still think that the Monascus anka Nakazawa et sato filament extract can effectively prevent and treat high fat+vitamin D
3The osteoporosis of rat.
3.3 the barrenwort alcohol extract is to high fat+vitamin D
3The osteoporosis rat preventive and therapeutic effect
By experimental result: the barrenwort alcohol extract is treated high fat+vitamin D
3Rat, experiment is compared with the model group rat after finishing, and serum TC, LDL-c obviously descend (P＜0.01), and HDL-c, HDL-c/TC, HDL-c/LDL-c obviously raise (P＜0.01), and the blood vessel elasticity fiber is looser, and atheromatous plaque forms less; With blood fat health group ratio, barrenwort alcohol extract group LDL-c obviously descends (P＜0.01), and HDL-c, HDL-c/TC obviously raise (P＜0.05), but proliferation of fibrous tissue companion punctate clacification, this possibly be since the blood fat health to the protective effect of blood vessel stronger due to.Result of experiment can know that the barrenwort alcohol extract treats high fat+vitamin D
3Rat is compared with the model group rat, and BMC increases, and bone mineralization strengthens, and the bone trabecula number increases, and explains that the barrenwort alcohol extract can increase the bone amount, has improved bone trabecular microstructure.Lumbar spine bmd is obviously increased, and the biomechanical property of bone makes moderate progress.So we think that the barrenwort alcohol extract can effectively prevent and treat high fat+vitamin D
3The osteoporosis of rat has the bone mineralising of promotion and osteoplastic effect.
3.4 Monascus anka Nakazawa et sato filament extract and barrenwort alcohol extract suspension are to high fat+vitamin D
3The osteoporosis rat preventive and therapeutic effect
Can know that by this result of experiment Monascus anka Nakazawa et sato filament extract and barrenwort alcohol extract suspension oral gavage are to high fat+vitamin D
3Rat, experiment is compared with the model group rat after finishing, and bone forms and bone mineralization strengthens, trabecular bone structure chap thickening, bone calcium and lumbar spine bmd increase, and the biomechanical property of bone makes moderate progress to a certain extent.Still think that Monascus anka Nakazawa et sato filament extract and barrenwort alcohol extract suspension can effectively prevent and treat high fat+vitamin D
3The osteoporosis of rat.
Embodiment two:
By the monascus hypha extract of the inventive method preparation,, be pressed into tablet by the traditional Chinese medicines production technology; Or be packed as capsule, or process granule, during electuary; Supply with osteoporotic animal by the clinical dosage of taking and take, provable its has tangible prevention effect to osteoporosis.
Embodiment three:
The monascus hypha extract that the present invention extracts as stated above; Add by 10% and 20% additive in the factory formula of biscuit; Produce the biscuit that contains the Monascus anka Nakazawa et sato filament extract, prove, this biscuit through zoopery; Have the osteoporotic effect of excellent prevention, can supply the patient who suffers from the osteoporosis disease to take as healthy food.
Embodiment four:
The monascus hypha extract that the present invention extracts as stated above; Add by 5%, 10% and 20% additive in the factory formula of beverage; Produce the beverage that contains the Monascus anka Nakazawa et sato filament extract, prove, this beverage through zoopery; Have the osteoporotic effect of excellent prevention, can supply the patient who suffers from the osteoporosis disease to use as healthy food.
Claims (6)
1. the red colouring agent for food, also used as a Chinese medicine liquid-state fermentation technology extracts the application of Monascus anka Nakazawa et sato filament extract remaining behind the monascorubin at preparation prevention of osteoporosis health food and medicine, and wherein the Monascus anka Nakazawa et sato filament preparation method of extract is: (1) get raw material such as rice with monascus through liquid deep layer fermenting after 7 to 10 days, the product liquid that the generation monascus ferments; Remove water partial pressure through squeezer, then, 60～70% alcohol extract monascorubins; Extracted the Monascus anka Nakazawa et sato filament of monascorubin, dried, pressed per kilogram and add 0.1～0.5 liter of 1% acetic acid; Under 25 ℃ of room temperatures, keep more than 24 hours,, extract twice by extraordinarily water extraction of gross weight 5～8; Each 1 hour, reclaim extracted twice liquid, the dried cream of simmer down to; Pulverize and be fine powder, room temperature preservation; Perhaps (2) are got raw material such as rice and after 7 to 10 days, are produced the product liquid of monascus fermentation through liquid deep layer fermenting with monascus, remove water partial pressure through squeezer, then; 60～70% alcohol extract monascorubins have extracted the Monascus anka Nakazawa et sato filament of monascorubin, dry; Press per kilogram and add 0.1～0.5 liter of 1% sodium acid carbonate, under 25 ℃ of room temperatures, keep more than 24 hours, by gross weight 5～8 extraordinarily water extract; Extract twice, each 1 hour, reclaim extracted twice liquid; The dried cream of simmer down to is pulverized and is fine powder, room temperature preservation; Perhaps (3) are got raw material such as rice and after 7 to 10 days, are produced the product liquid of monascus fermentation through liquid deep layer fermenting with monascus, remove water partial pressure through squeezer, then; 60～70% alcohol extract monascorubins have extracted the Monascus anka Nakazawa et sato filament of monascorubin, dry, by gross weight 5～8 extraordinarily water extract; Extract twice, each 1 hour, reclaim extracted twice liquid; The dried cream of simmer down to is pulverized and is fine powder, room temperature preservation.
2. application as claimed in claim 1, the form of medicine are tablet, capsule, granule, electuary.
3. application as claimed in claim 1, wherein health food is meant biscuit, beverage.
4. application as claimed in claim 3, the Monascus anka Nakazawa et sato filament extract is 5～30% at the addition of biscuit.
5. application as claimed in claim 3, the Monascus anka Nakazawa et sato filament extract is 5～20% at the addition of beverage.
6. according to claim 1 or claim 2 application is characterized in that said Monascus anka Nakazawa et sato filament extract mixes use with the barrenwort alcohol extract.
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| CN102813692A (en) * | 2012-05-24 | 2012-12-12 | 广东医学院 | Production method and clinical application of healthcare foods and medicines having monoamine oxidase B inhibition function |
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| CN101579388A (en) * | 2008-05-15 | 2009-11-18 | 北京北大维信生物科技有限公司 | Medicinal composition for treating osteoporosis and preparation method thereof |
| CN101836725A (en) * | 2010-01-01 | 2010-09-22 | 广东医学院 | Application of red rice residue product in preparing health-care food for lowering blood fat |
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