CN102366431A - Application of ginseng flower in preparing medicinal preparation for controlling osteoporosis - Google Patents
Application of ginseng flower in preparing medicinal preparation for controlling osteoporosis Download PDFInfo
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Abstract
The invention relates to application of an extract of ginseng flower in preparing a medicinal preparation for controlling osteoporosis. Ginseng flower is bud of Panax.ginseng C.A.Meyer and is subjected to processes of crushing, immersion, filtration, condensation, drying and the like to prepare extract or made into a preparation for clinical application according to a pharmaceutic process; the preparation of ginseng flower has an obvious preventing effect on osteoporosis of rats caused by ovariectomization and a good curative effect on aging osteoporosis of rats caused by usage of D-galactose. The invention prompts that the preparation of ginseng flower has an obvious preventing and treating effect on postmenopausal osteoporosis and senile osteoporosis and can be used as a medicine or a functional foodstuff for preventing and treating senile osteoporosis of human beings.
Description
Technical field the present invention relates to the application of Flos Ginseng's extract in the pharmaceutical preparation of preparation Prevention and Treatment of Osteoporosis.
The background technology osteoporosis is because the bone amount reduces, the fine structure of bone is degenerated and the fragility of bone increases; Cause the biomechanical strength of bone to descend; Be difficult to carry the load of original or relative increase; Capillary fracture or complete fracture can quietly take place, have a strong impact on old people's quality of life, bring very big burden for patient, family and society.The epidemiological study of osteoporosis shows, women's menopause in 49 years old can be suffered from the bone amount in 52--60 year and reduced, and 62--76 year can suffer from osteoporosis, and 72--86 year can be suffered from serious osteoporosis.Except that primary osteoporosis; Secondary osteoporosis; Particularly corticosteroid osteoporosis also faces stern reality, in the patient of the chronic disease of accepting corticosteroid treatment, finally also has 30～50% to develop into osteoporosis; More serious is to cause femur head necrosis and Fracture of femur, is the iatrogenic disorder problem that can not be ignored.At any time development and national economy, the raising of living standards of the people, the population life-span prolongs gradually, and following society will become senile society.China more than 60 years old aging population risen to 1.34 hundred million people, according to international standard, China has got into aged society.If add the person in middle and old age's population below 60 years old, its quantity is more considerable.As the osteoporosis and the osteoporotic fracture of one of Senile disease, increasing people is deeply hurt, and become the serious social problem of worldwide concern, China has classified it as one of Senile disease of three big emphasis tackling key problem researchs.WHO works up the global strategy target to osteoporosis; Improve the whole world to osteoporotic diagnosis and processing; Each university, institute, drugmaker one after another first mate's degree increase funds to quicken the research and development of osteoporosis new drug; Therefore preventing and treating senile osteoporosis is to guarantee that people are healthy, the research topic that ten minutes is urgent of improving the quality of living.
Osteoporosis is a kind of disease of multi-pathogenesis; Its basic pathology mechanism is that defective appears in bone resorption and osteoplastic balance in the bone metabolism process, causes the intravital alcium and phosphor metabolization imbalance of people, bone density is reduced gradually and cause clinical symptoms, its cause of disease is main and the age; Endocrine regulation; Calcium absorption is bad, and limbs are useless to be used and immunity, and factors such as nutrition, heredity are relevant.At in recent years 20 years, osteoporotic Drug therapy strategy and approach mainly were two big types, and one is the medicine of anti-bone resorption, and two for facilitating the bone medicine.Being anti-bone resorption medicine more than 90% in the clinical use, like controversies in hormone replacement in the elderly, is the academic therapy of treatment postmenopausal osteoporosis.But estrogen causes the danger of genital system tumor and is affirmed that the U.S. has declared that estrogen is one of carcinogen; The diphosphonic acid salt can effectively be treated bone loss due to the bone resorption, but the prolonged application side effect is big, also is difficult for as prophylactic; Calcitonin is remarkable to dissolving bone property osteodynia and osteoporosis effect, but treatment needs individuation, also receives the route of administration restriction, and costs an arm and a leg; The estrogenic agents of paying close attention at present (ERMs) is blocked the stimulation to reproductive system owing to having kept to the estrogen-like effects of skeleton, has advantage, has 2-3 new drug and gets into the clinical III phase and test (U.S.).All these anti-bone resorption agent have all suppressed because of bone resorption increases the bone formation that coupling joins, and the conversion ratio of old bone and new bone is reduced, and prolonged application influences the quality and the mechanical property of bone.At present short bone formation medicine becomes the research focus gradually; Short chain parathyroid hormone (PTH) is very significantly short bone formation medicine; It is clinical also to get into the II phase in the U.S.; But experimental studies results finds that PTH can increase cortical bone porous (because of increasing the absorption of cortical bone Haversian canal simultaneously), reduces the late result of fracture and does not affirm [4]; Active VitD prolonged application is prone to cause hypercalcemia and urine calcium disease; Fluoride is the short bone formation medicine that early is used for osteoporosis, but it is prone to make osteoid to increase and with bone mineralising defective, has limited in wide clinical application, and the safety range that mainly comes from it is narrow, and side effect is big, and curative effect needs further textual criticism.To the treatment of senile osteoporosis, also there is not excellent drug at present.
This project team is through a large amount of animal experiment studies and screening, the dry root of once having found Chinese medicine Araliaceae Radix Ginseng (Panax ginseng C.A.Mey) with and the extract Ginsengdiol histsaponin of dry root; The extract stem and leaf of Radix Ginseng total saponins of Stem and leaf of Radix Ginseng has the osteoporotic effect of tangible control; Applied for national inventing patent in 2000; Number of patent application is: ZL00104283.1, and this patent was obtained the authorization in 2004, and certificate of invention number is: No. 143882.Through further investigation for many years, we find that the preparation that alabastrum of Radix Ginseng is processed also has the osteoporotic effect of good control, and particularly treatment has tangible advantage to senile osteoporosis.
The Flos Ginseng is that Radix Ginseng (Panax.ginseng C.A.Meyer) is the alabastrum of Araliaceae Radix Ginseng, has another name called " SHENCAO flower ", picks in the Radix Ginseng bud that a bud just ready to burst, bakes naturally to form.The Flos Ginseng grows to 4 years blooms soon, and every Flos Ginseng only opens a Xiao Hua every year, and per 60 jin of Radix Ginsengs only can be adopted to such an extent that one or two ginseng is spent, and the title of " green gold " is arranged.The Flos Ginseng is contained 20 kinds of Saponin active substances, 17 seed amino acids, 11 kinds of trace element, three kinds of active anticancer selenium and crude protein etc.In recent years research thinks that its medical value is better than Radix Ginseng; Research to Flos Ginseng's composition; Find the contained various compositions of alabastrum of Radix Ginseng reach 300 surplus kind, alabastrum of Radix Ginseng contains the whole compositions of Radix Ginseng root, and monomer ginsenoside-Rms7cd that the Radix Ginseng root does not have contains in alabastrum of Radix Ginseng only and deposits; Alabastrum of Radix Ginseng also contains the ginseng polypeptide that does not have in the Radix Ginseng, and content is high; The ginsenoside content is higher 5 times than Radix Ginseng, and especially saponin monomer Re exceeds 10 times than Radix Ginseng.Total Saponin in the alabastrum of Radix Ginseng not only content is high, and equilibrium is also very desirable.Having the Rbc system of sedation effect and having excitation Rgl system respectively accounts for half the, balanced very perfect.The ginsenoside Re has the secretion of the cortin of succenturiate kidney of enhancing, promotes that DNA is synthetic and makes the macrophage activation effect.Rms7cd and ginseng polypeptide delaying sanility, allaying tiredness, raising immunity, prevention various diseases.Reported that alabastrum of Radix Ginseng mainly has following effect: 1. slow down aging, allaying tiredness improves insomnia; Improve memory and sexual function, eliminate the aching and limp and pain of waist, head, shoulder, the prevention radiation; 2. raising immunity, infectious diseases such as preventing cold, pneumonia, nephritis, hepatitis; 3. prevention of arterial sclerosis, coronary heart disease, apoplexy, hypertension, hypotension and the headache that causes thereof, tinnitus; 4. improve climacteric syndrome, regulating menstruation, calm the nerves, remove agitation; 5. prevent diabetes, hypotension and the giddy that causes thereof, dim eyesight; 6. the face red spot that disappears, it is beautiful to renew one's youth.About Flos Ginseng's pharmacological research, existing following report:
1. to the influence of cardiovascular system: give mouse gavaging Flos Ginseng Saponin; Dosage is respectively 12.5mg/kg, 25mg/kg and 50mg/kg; Continuous 10d, the cAMP content that records 3 dose groups mouse cardiac muscles all is significantly increased than normal saline group mouse cardiac muscle cAMP content and strengthens with dosage increase effect, and Flos Ginseng's Saponin also makes mouse cardiac muscle cAGP content increase; Except the 12.5mg/kg dose groups, all the other respectively organize significant difference.3 dosage increase, and except the 12.5mg/kg dose groups, all the other respectively organize significant difference.The cAMP/cAGP ratio of 3 dose groups also increases with dosage and improves.
2. antiulcer action: Flos Ginseng's Saponin 60mg/kg intramuscular injection is to pylorus ligation; Experimental gastric ulcer animal model due to reserpine, the aspirin all has inhibitory action, and can obviously suppress rat gastric secretion, reduction Both of gastric acidity and protease activities.To histamine and the rat and the contraction of guinea pig in vitro enterospasm property that cause of phatidylcholine, the part antagonism is arranged also.
3. antitumor action: the in vitro tests of Flos Ginseng's Saponin can improve mice spleen spleen natural killer cell (NKC) activity, and can exist at canavaline-A (Con-A) and induce biogamma (γ-IFN) and interleukin-2 (IL-2) under the situation; The NKC that can improve normal mouse in vivo in the test is active, and kind of the NKC that plants tumor mice ability active, that produce γ-IFN and IL-2 is enhanced.Present Flos Ginseng's Saponin NKC-IFN-IL-2 is regulated the positive control of net, show that Flos Ginseng's Saponin possibly bring into play tumor-inhibiting action through inducing IFN, through promoting the active and killing tumor cell more effectively of NKC.After Flos Ginseng's glycol glycoside and mouse hydroperitoneum type reticulosarcoma (ARS) the cell co-cultivation, suppress the synthetic and karyokinesis of its DNA, and present reverse to a certain degree.
3. delaying senility function: Flos Ginseng's liquid glucose Apis of feeding, can make body constitution obviously strong, the working time is long, and efficient is high, and foraging quantity obviously increases.And the bee colony mortality rate is low, but than matched group life-saving 10-15d.Lumbar injection Flos Ginseng Saponin 60mg/kg, ability significant prolongation anoxia mice life span and mice swimming time have tangible antioxygen and antifatigue effect.
Flos Ginseng's Saponin reaches rat urine amount is obviously reduced, and presents significant antidiuretic activity.
So far do not see bibliographical information about Flos Ginseng's prevention and treatment osteoporosis.
Summary of the invention task of the present invention is to make preparation to the extract of Flos Ginseng and active ingredient thereof with the technology of science; Make said preparation be used for human Prevention and Treatment of Osteoporosis; Our research has proved that said preparation can stop loss of bone; Can promote the formation of new bone again, new medicine is provided for solving an osteoporotic difficult problem.
Flos Ginseng's preparation can be through following prepared:
(1) gets the Flos Ginseng, pulverize, add 15 times of boiling water and soaked 30 minutes, filter, keep filtrating; Filtering residue adds 15 times of boiling water again and soaked 30 minutes, filters, and keeps filtrating; Filtering residue adds 10 times of 60% soak with ethanol 24 hours, after, heating extraction 2 hours is filtered; Reclaim ethanol, must filtrate, merge filtrating three times, simmer down to extractum or drying; Become every to contain crude drug 0.5 gram by the tablet prepared, in addition, also can process other preparations that can supply clinical practice by pharmaceutics technology.
(2) get the Flos Ginseng, pulverize, carry out micronization, add 15 times of boiling water and soaked 30 minutes, filter, keep filtrating with the Baily pulverizing mill; Filtering residue adds 15 times of boiling water again and soaked 30 minutes, filters, and keeps filtrating; Filtering residue added 10 times of 60% soak with ethanol after 24 hours, and heating extraction 2 hours is filtered, and reclaims ethanol; Must filtrate, merge filtrating three times, dense extractum or the drying of being condensed to; Become every to contain crude drug 0.5 gram by the tablet prepared, in addition, also can process other preparations that can supply clinical practice by pharmaceutics technology.
(3) get the Flos Ginseng, pulverize, carry out micronization, add 15 times of 20 ℃～25 ℃ distilled water immersions 30 minutes, filter, keep filtrating with the Baily pulverizing mill; Filtering residue adds 15 times of 20 ℃～25 ℃ distilled water immersions 30 minutes again, filters, and keeps filtrating; Filtering residue adds 10 times of distilled water, transfers PH to 1 with hydrochloric acid; Reacting by heating 2 hours is put coldly, adds alkali and transfers PH to 7; Filter, must filtrate.Merge filtrating three times, simmer down to extractum or drying become every to contain crude drug 0.5 gram by the tablet prepared, in addition, also can process other preparations that can supply clinical practice by pharmaceutics technology.
(4) get the Flos Ginseng, pulverize, add 15 times of 70% soak with ethanol 24 hours, heating extraction 2 hours is filtered, and must filtrate.Filtering residue adds 40% soak with ethanol 24 hours again, and heating extraction 2 hours is filtered, and must filtrate.Merge filtrating twice, simmer down to extractum or drying become every to contain crude drug 0.5 gram by the tablet prepared, in addition, also can process other preparations that can supply clinical practice by pharmaceutics technology.
(5) get the Flos Ginseng, pulverize, carry out micronization with the Baily pulverizing mill, add 15 times of 70% soak with ethanol 24 hours, heating extraction 2 hours is filtered, and must filtrate.Filtering residue adds 40% soak with ethanol 24 hours again, and heating extraction 2 hours is filtered, and must filtrate.Merge filtrating twice, simmer down to extractum or drying become every to contain crude drug 0.5 gram by the tablet prepared, in addition, also can process other preparations that can supply clinical practice by pharmaceutics technology.
The Flos Ginseng's extractum or the tablet of above-mentioned prepared, through the osteoporosis rat Research of Animal Model for Study, the result shows: Flos Ginseng's tablet of above five prepared all has and prevents the osteoporotic effect of laboratory animal preferably.These tablets, the osteoporosis rat that removal ovary is caused and cause rat aging property osteoporosis that better curative effect is all arranged with the D-galactose.The D-galactose causes old and feeble animal model by the human antidotal research of extensive conduct; The D-galactose causes rat aging property osteoporosis and human senile osteoporosis quite similar; Flos Ginseng's preparation has tangible prevention and therapeutical effect to this model, has shown that Flos Ginseng's preparation can be used as the medicine or the functional food of human senile osteoporosis prevention and treatment.
More than by the preparation of Flos Ginseng preparation, also can be made into clinical other acceptable preparations, any as in tablet, pill, electuary, granule, capsule, injection, drop pill, the preparation for external application to skin supplies clinical practice.
The specific embodiment:
Embodiment one:
The osteoporotic experimentation of Flos Ginseng's extractum antagonism removal ovary female rats
1 experiment material and method
1.1 main medicine
1.1.1 Flos Ginseng's extractum: the extractum that the production technology that provides by this project obtains; 500mg/ grain (being equivalent to 900mg extractum), high dose is got extractum 29.16g, and adding distil water is diluted to 600ml; By the rat oral gavage amount is 10ml/kg/d and adult's dosage, and conversion dosage is 0.486g/kg/d.High dose=low dosage * 5.
1.1.2 livial: Ou Jianong pharmaceutical Co. Ltd in Nanjing produces, lot number LH-0502-10,2.5mg/ sheet; Getting two clays into power earlier; Adding dissolved in distilled water and become 222ml solution, is 10ml/kg/d and adult's dosage by the rat oral gavage amount, and conversion dosage is 0.225mg/kg/d.
1.1.3 quadracycline: powder, new Asia, Shanghai pharmaceutical factory produces, and keeps in Dark Place.With the dilution of 0.9% sodium chloride solution, be mixed with 2.5% solution before using, join existing usefulness at present, keep in Dark Place.Give the rat neck subcutaneous injection, consumption is 1ml/kg.
1.1.4 calcein (calcein): powder, Sigma Chemical Co., USA product.With 2%NaHCO3 normal saline 100ml, magnetic agitation slowly adds calcein powder 0.5g simultaneously, treats to dissolve fully the back and filters with microfilter, is 0.5%calcein solution, puts 4 ℃ of refrigerators and keeps in Dark Place subsequent use at the black out environment.Give the rat neck subcutaneous injection, consumption is 1ml/kg.
1.2. main agents and instrument and equipment
Slightly
1.3 experimental technique
53 of 3 monthly age of SPF level SD male rats, body weight during experiment (201 ± 20) g; 5 monthly ages of SPF level are 66 of copulation SD female rats not, body weight during experiment (269 ± 28) g.(the animal quality certification number is provided: 2006A015) by Guangdong Medical Lab Animal Center; The raising condition: raise indoor temperature and keep 24 ℃～28 ℃, humidity is 50%～60%, special chamber sub-cage rearing, two in every cage, the next day change bedding and padding, freely take the photograph water, ingest (standard feed that the court's animal center provides), weigh once weekly.
1.3.1 experimental design and grouping
50 5 monthly age SPF level SD rats are divided into 6 groups at random by the body weight principle of reciprocity, and each treated animal number and medication are seen table 3.1, every day gastric infusion, successive administration 12w:
Table 3.1 experimental design and grouping
All male rats are put to death preceding 13,14 days subcutaneous injection 25mg/kg quadracyclines respectively once, in putting to death preceding 3,4 two days subcutaneous injection calcein 5mg/kg respectively once, two fluorescent labelinies 10 days at interval.Body weight of weighing weekly, the feeding of freely drinking water.After experiment finishes, with executions (to reduce erythrocyte in the bone marrow, the eliminating interference makes osteocomma be easier to observation analysis) of thoroughly drawing blood of right ventricle behind 3% pentobarbital sodium (1.5ml/kg) the row intraperitoneal injection of anesthesia.Detect index: 1. left side tibia epimere and fifth lumbar vertebra is processed the undecalcified bone slice, the stage casing bone is processed undecalcified bone abrasive disc, carry out osseous tissue morphometry parameter detecting.2. the left side femur is measured the content of its bone hydroxyproline, bone mineral element.3. the right side femur carries out bone biomechanical mensuration.
In the experimentation, weigh weekly once, water is can't help in fasting the previous day of weighing, and weighs before the administration in second day, and data typing computer is adjusted dosage then and carried out administration.
1.3.2 ovariectomized rats operation method
After the rats by intraperitoneal injection pentobarbital sodium 40mg/kg anesthesia, be the center with 1cm place under the midaxillary line waist rib, cropping diameter 2cm after sterilizing successively with iodine tincture, ethanol, carries out removal ovary and performs the operation.
To between strand each digit of upper limb, taught skin is done 0.8～1cm otch along longitudinal axis to operative incision under rib, and last stricture of vagina pincers lead and carry otch about otch.In straight mosquito forceps passivity separate subcutaneous superficial fascia organized layer, folder is carried the flesh layer of seeing pink light, clamps on the offside, scalpel is made the longitudinal axis and is cut the flesh layer, can reach the abdominal cavity.Change stricture of vagina pincers clamp and lead elevator layer otch, expose the abdominal cavity, (mature ovarian is a pale red to have led intraperitoneal tissue searching ovary gently with tweezers; Oval; There is the irregular nodular pattern follicle on the surface, for terminal, pink tubulose uterus connects, more white adipose frenulum is arranged on every side).After finding ovary, clamp zygote palace on it is following.Pincers are gutstring ligation uterus down, and cutter eliminates pincers and goes up ovary tissue, and remaining tissue is sent into intraperitoneal, closes abdomen, sews up flesh layer and cortex.Tetracycline eye ointment is coated with eyes, puts back in the cage, lies on one's side.Sham-operation (sham) process not ligation uterus and non-spay, surplus and last same.Postoperative places warm environment, and nurse is observed.
1.3.3 rat bone fluorescence labeling method
All animals respectively gave cervical region subcutaneous injection quadracycline (25mg/kg) in preceding the 14th, 13 day in the experiment end; As the fluorescent labeling first time; Respectively gave cervical region subcutaneous injection calcein (5mg/kg) in preceding the 4th, 3 day as the fluorescent labeling second time in the experiment end; Forming yellow and green two fluorized markings at bone surface, be 10 days twice fluorescent labeling blanking time, can dynamic observe osteoplastic situation during the double injection.Under fluorescence microscope, the tetracycline fluorescence that is deposited on bone surface is yellow, and the calcein fluorescence that is deposited on bone surface shows green, it is carried out corresponding measurement can dynamic observe the sedimentary speed of bone surface mineralising, the osteoplastic situation of reflection osteoblast.
1.1.4 rat bone specimen sampling method
Take out rat bilateral tibial and femur and fifth lumbar vertebra rapidly after getting blood, separate Ex-all and adhere to muscle and connective tissue, left side femur constant temperature is baked to constant weight and gets key weight, is used for bone physical index and bone biochemical indicator and detects.The left side tibia cuts metaphysis with the low speed saw along frontal plane, to expose medullary cavity, walks crosswise sawed-off tibia in 1/3 place, tibia upper end again, and this is the tibia upper end.The tibia of remainder part walked crosswise in tibia and fibula junction cut, do stage casing bone detection.Fifth lumbar vertebra is fallen 1/3 along the sagittal plane length sawing, remove unnecessary osseous tissue, detect to make lumbar vertebra.
3.1.5 undecalcified bone slice method for preparing
Press the embedding of undecalcified bone slice method for preparing to tibia and lumbar vertebra; The BIAO and BEN that embedding is good is cut into two kinds of thickness bone slices of 5 μ m and 8 μ m; 5 μ m thin slices are used for Masson-Goldner Trichrome method and dye, and the direct mounting that do not dye after 8 μ m sheets take off and mould is observed.
1.1.6 cortical bone undecalcified bone abrasive disc method for preparing
Abrasive disc is that middle tibia is sawn into the thick osteocomma of 100 μ m with veneer sawing machine earlier, is separated into effective osteocomma fully, the error 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 tibia 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 40 μ m-50 μ m, carry out ethanol gradient dehydration: 70%-70%-95%-95%-100%-100% with coarse glass again, cross twice of xylene.Transparent back resin mounting, the parameters of measurement cortical bone.
1.1.7 the measuring method of osseous tissue morphometry
Measuring method according to the osseous tissue morphometry; Directly measure each concrete data of osseous tissue with semi-automatic image digitizer; And calculate through international formula these concrete data to it; Obtain bone amount, bone structure, bone formation and bone resorption morphology parameter respectively, be used to analyze and estimate the effect of medicine.The parameter of osseous tissue morphometry comprises:
Calculate the gained parameter and can be divided into static parameter and dynamic parameter.
1. static parameter: be used for estimating the medical treatment effect, describe what and bone trabecular morphosis of bone amount.
Bone trabecula area percent (%Tb.Ar): phalanges girder area accounts for the percentage rate of osseous 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 width (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 number (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.
Cortical bone area (Ct.Ar): what of reflection cortical bone bone amount are the poor of stage casing osseous tissue cross section and medullary cavity area.
Cortical bone area percent (%Ct.Ar): reflection cortical bone proportion in the total osseous tissue area of stage casing bone cross section.
Medullary cavity area percent (%Ma.Ar): reflection medullary cavity proportion in the total osseous tissue area of stage casing bone cross section.
2. dynamic parameter: comprise bone formation parameter and bone resorption parameter,, can understand the amount and the speed of bone surface mineralising, and explain the reason that static parameter changes through dynamic parameter.
A bone formation parameter
Fluorescent perimeter percent (%L.Pm): reflect that the bone girth that carries out mineralising accounts for the percentage rate of bone surface overall circumference, also reflects osteoblastic quantity.
Bone mineralising deposition (MAR): refer to the width of 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 ratio of the bone trabecula gross area, represents the active degree of bone formation and bone conversion.This more parameters is high, and the conversion of expression bone is also high more, is the osteoplastic important indicator of reflection.
Bone formation rate (BFR/TV): the annual new bone trabecula area that forms accounts for the percentage ratio of osseous 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 just to provide the required bone trabecula of more new bone formation surperficial; The 2nd, bone formation active degree, bone formation are active more, and the new bone that on unit length, forms is just many more.Though so bone formation is very active sometimes, 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.
B bone resorption parameter
Osteoclast number (Oc.N): the osteoclast number on the unit bone trabecula area, reflect the bone resorption situation relevant with osteoclast.
Osteoclast girth percentage rate (%Oc.S.Pm): the bone trabecula girth that osteoclast covers accounts for the percentage ratio of bone trabecula total surface girth, and the osteoclast number what not only reflect, also reflects the active degree of osteoclast size and bone resorption thereof.
The cortical bone lining endothelium absorbs girth percentage rate (%E-L.Pm): the girth of cortex internal skeleton face Howship lacuna, the bone resorption function of reflection internal skeleton face osteoclast.
1.1.8 the assay method of femur weight, width and length
(1) assay method of femur dry weight
Get the left side femur, muscle and the soft tissue on the bone adhered in Ex-all, claims to toast 72h to constant weight in 80 ℃ of baking boxs 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.
(2) assay method of femur length
Hand-held rat femur is bent face in the face of femur, uses slide gauge to record the distance that greater trochanter of femur and neck of femur meet minimum point arrive fossa intercondylaris femoris and is femur length (BL).
(3) assay method of femur width
Forefinger and thumb block lateral condyle resected femur face in the wrong in femoral head and the femur, and the third trochanter that blocks femoral curvatures with the slide gauge level comes downwards to femoral shaft handing-over part, and institute's value is femur width (BD).
Because the growth of the macroscopic view of bone comprises the long and long thick of bone, therefore measures the overall growth situation that femur length and width can embody bone.
1.1.9 the assay method of femur calcium, phosphorus and hydroxyproline content
The muscle and the fascia of left side femur are rejected totally, measure left side femur weight, width and length, after precision was weighed, 80 ℃ of baking 72h were to constant weight, and precision is weighed and record once more.Place 10ml ampere bottle to add 6mol/LHCL 5ml every part of bone specimen subsequently, alcohol blast burner is burnt and is taken out filtration after bottleneck seals back 108 ℃ of baking box constant temperature digestion 24h.Filtrating is divided into two parts, a with 1: 200 dilution back concentration with ICP appearance (inductance coupled plasma direct-reading spectrometer) mensuration bone Ca, bone P, calculate the content of bone Ca, bone P according to standard substance concentration and diluted sample multiple.Another part filtrating, femur were with dilution in 1: 10, and adjust pH to 6～7 backs calculate the content of hydroxyproline then according to the absorbance of hydroxyproline determination kit method with hydroxyproline in the UV-752 ultraviolet spectrophotometer working sample according to formula.
1.1.10 bone biomechanical detects
During test, with the femur normal temperature unfreezing of-20 ℃ of preservations, normal saline is wet again.Analyze the biomechanical property of right side femur (three-point bending experiment) with 858Mini Bionix type material test macro.During measurement, rat right side femur is placed on the MTS testing machine, 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 etc.The observation parameter of bone biomechanical is following:
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 (greatest force that bone can bear in elastic range); Maximum load (greatest force of being born before the bone fracture); Breaking load (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 osseous tissue.
Flexional: bone specimen is out of shape the energy of required absorption.
2 experimental results
2.1. Flos Ginseng's extractum is to the influence of removal ovary female rats body weight
Flos Ginseng's extractum is seen table 2.1 to the influence of the per two all body weight of removal ovary rat:
The influence
that table 2.1 Flos Ginseng extractum changes the removal ovary rat body weight
(
*P＜0.05，
**P＜0.01vs?Sham；
△P＜0.05，
△△P＜0.01vsOVX；
#P＜0.05，
##P＜0.01vs?LV)
Result by table 2.1 is visible, compares with the Sham group, and OVX group ovariectomized rats is after two weeks, and body weight begins to increase, and when experiment proceeded to for the 4th, 6,8,10,12 weeks, weight increase is (P＜0.05) more obviously, pointed out removal ovary that rat body weight is obviously increased.After using the LV treatment, the 2nd week of removal ovary rat body weight begins to alleviate (P＜0.05), and the prompting livial can suppress the increase of removal ovary rat body weight.GF (L), GF (H) do not have obvious influence (P＞0.05) to the removal ovary rat body weight; Compare with the LV group, GF (H) group rat body weight all is higher than LV group (P＜0.05), and the do not have obvious impact effect of high dose Flos Ginseng extractum to the removal ovary rat body weight is described.
2.2 Flos Ginseng's extractum is to the influence of removal ovary rat tibia epimere morphometry static parameter
Flos Ginseng's extractum is visible to removal ovary rat tibia epimere osseous tissue morphological observation, the bone trabecula of sham operated rats rat tibia epimere appear structure closely, even thickness, seriality be good, and no significant difference.The trabecular bone structure of OVX model group animal is obviously sparse, tiny, large stretch of no bone trabecula bone marrow district occurs.The bone trabecula of livial treated animal increases than model group, and seriality is also recovered to some extent, but not statistically significant.The trabecular bone of Flos Ginseng's extractum low dose group, Flos Ginseng's extractum high dose group animal is fine and close, thick, long, and is evenly distributed, and seriality is fabulous, than model group tangible improvement has been arranged.
Flos Ginseng's extractum is seen table 2.2 to the influence of static parameter %Tb.Ar (bone trabecula area percent), Tb.Th (bone trabecula thickness), Tb.N (bone trabecula quantity), Tb.Sp (bone trabecula separating degree) and bone resorption parameter Oc.N in the removal ovary rat tibia epimere bone histomorphometry (unit bone trabecula area osteoclast number), %Oc.Pm (osteoclast girth percent), Oc.Nmm (every millimeter osteoclast number).
Table 2.2 Flos Ginseng extractum is to the influence
of removal ovary rat tibia epimere static parameter
(
*P＜0.05，
**P＜0.01vs?Sham；
△P＜0.05，
△△P＜0.01vsOVX；
#P＜0.05vs?LV)
Can be known by table 2.2: compare with the Sham group, the bone trabecula quantity of OVX group obviously reduces, sparse, the fracture of structure; %Tb.Ar, Tb.N have reduced by 63.9% (P＜0.01), 60.5% (P＜0.01) respectively, and bone resorption parameter Oc.N/mm increases by 143.2% (P＜0.05); Explain that removal ovary has made the bone trabecula of rat spongy bone significantly reduce, microstructure is loose degenerates, and osteoclast is had facilitation.Compare with OVX group, the bone trabecula showed increased of GF (H) group, increase slightly, seriality is good; The Oc.N/mm of LV has reduced by 68.2% (P＜0.05); The Tb.N of GF (L) increases by 47.1% (P＜0.05); The %Tb.Ar of GF (H), Tb.Th have increased by 64.8% (P＜0.05), 27.1% (P＜0.01) respectively; Oc.N/mm has reduced by 65.4% (P＜0.05), though Tb.Sp descends not statistically significant.Though and livial has suppressed the absorption of osteoclast, the bone loss that removal ovary is caused makes moderate progress, and does not have significant change; Flos Ginseng's extractum that high and low dose is described all can effectively prevent bone loss and the microstructural destruction of removal ovary to the rat tibia epimere, and livial can not effectively resist the variation of the rat tibia epimere that removal ovary causes.Bone resorption parameter Oc.N, %Oc.S.Pm and Oc.Nmm have downward trend, but not statistically significant points out this medicine preventive dose that the active degree of osteoclast number and bone resorption is not had remarkable inhibitory action.
2.3 Flos Ginseng's extractum is to the influence of removal ovary rat tibia epimere morphometry dynamic parameter
Flos Ginseng's extractum is seen table 2.3 to the influence of dynamic parameter %L.Pm (fluorescent perimeter percentage rate) in the removal ovary rat tibia epimere osseous tissue morphometry, MAR (bone mineralising deposition) and three kinds of bone formation rate index BFR/BS, BFR/BV, BFR/TV:
Table 2.3 Flos Ginseng extractum is to the influence
of removal ovary rat tibia epimere dynamic parameter
(
*P＜0.05vsSham；
△P＜0.05，
△△P＜0.01vsOVX)
Can know by table 2.3, compare that the fluorescence of OVX group rat spongy bone is intensive, abundant with the Sham group; Except that the BFR/TV not statistically significant; %L.Pm, MAR, BFR/BS, BFR/BV have increased by 139.4%, 39.1%, 249.5%, 303.8% (P＜0.05) respectively; The prompting removal ovary can obviously promote bone formation and the bone mineralization of rat; Bone turnover rate is high, bone is built be in positive balance again and is caused bone loss.Compare with the OVX group, the dynamic parameter index of prophylactic agent group is except that the BFR/TV not statistically significant, and %L.Pm, MAR, BFR/BS, the BFR/BV of LV group have reduced by 81.6%, 25.8%, 85.0%, 86.4% (P＜0.05) respectively; The %L.Pm of GF (L), MAR, BFR/BS, BFR/BV have reduced by 60.8%, 21.9%, 68.6%, 72.6% (P＜0.05) respectively; The %L.Pm of GF (H), MAR, BFR/BS, BFR/BV have reduced by 75.3%, 44.5%, 85.6%, 88.8% (P＜0.01) respectively.Explain that the prophylactic agent group suppresses the tibia epimere bone mineralising bone formation increase that removal ovary causes to some extent.
2.4 Flos Ginseng's extractum is to the influence of removal ovary middle tibia cortical bone static parameter
Flos Ginseng's extractum is seen table 2.4 to the influence of removal ovary rat tibia stage casing cortical bone static parameter Ct.Ar (cortex area), %Ct.Ar (cortex area percentage rate), %Ma.Ar (bone marrow area percentage rate):
Table 2.4 Flos Ginseng extractum is to the influence
of removal ovary rat tibia stage casing cortical bone static parameter
(
*P＜0.05vs?Sham；
△△P＜0.01vs?OVX；
#P＜0.05vsLV)
Can know by table 2.4, with Sham group relatively, the %Ct.Ar of OVX group rat 4.1% (P＜0.05) that descended, %Ma.Ar has increased by 16.3% (P＜0.05); The prompting removal ovary diminishes rat cortical bone area, and it is big that medullary cavity becomes, and explains that removal ovary can cause rat cortical bone bone loss.With OVX group relatively, the LV group %Ct.Ar, %Ma.Ar change not quite, the prompting livial is little to the cortical bone bone amount effect of removal ovary rat.GF (L), GF (H) group makes %Ct.Ar increase by 4.6% (P＜0.01) and 4.3% (P＜0.01) respectively, makes %Ma.Ar descend respectively 14.8% (P＜0.01) and 14.0% (P＜0.01); Prompting GF increases removal ovary rat cortical bone area, and medullary cavity is dwindled, and explains that the cortical bone bone amount that Flos Ginseng's extractum can effectively resist the removal ovary rat obviously reduces.Compare with the LV group, the effect of GF (L) is better than LV (P＜0.05).
2.5 Flos Ginseng's extractum is to the influence of removal ovary rat cortical bone dynamic parameter
Flos Ginseng's extractum is seen table 2.5 to %E-L.Pm (fluorescence percent), E-MAR (mineralising deposition), the E-BFR/BS (bone formation rate) of %P-L.Pm (fluorescence percent), P-MAR (mineralising deposition), P-BFR/BS (bone formation rate) and the perimyelis of removal ovary rat cortical bone dynamic parameter periosteum.
Table 2.5 Flos Ginseng extractum is to the influence
of removal ovary rat tibia stage casing cortical bone dynamic parameter
(
**P＜0.01vsSham；
△P＜0.05，
△△P＜0.01?vsOVX)
Can know by table 2.5; Compare with the Sham group; OVX group rat tibia stage casing cortical bone is except that %E-L.Pm has increased by 273.7% (P＜0.01), and other dynamic parameters of interior adventitia do not have significant change, and the prompting removal ovary can strengthen the bone formation and the mineralization of rat tibia stage casing cortical bone inner membrance.Compare with the OVX group; The %E-L.Pm of LV group reduces by 59.2%; The %E-L.Pm of GF (L), GF (H) group descend respectively 43.7% (P＜0.05) and 69.0% (P＜0.01); But other dynamic parameters do not have significant change, explain that livial and Flos Ginseng's extractum have resisted the bone formation of removal ovary rat tibia stage casing cortical bone inner membrance and the increase of mineralising, do not have obvious influence to periosteum.
2.6 Flos Ginseng's extractum is to the influence of removal ovary rat fifth lumbar vertebra morphometry static parameter
Flos Ginseng's extractum is seen table 2.6 to the influence of static parameter %Tb.Ar (bone trabecula area percent), Tb.Th (bone trabecula thickness), Tb.N (bone trabecula quantity), Tb.Sp (bone trabecula separating degree) in the removal ovary rat fifth lumbar vertebra meterological:
Table 2.6 Flos Ginseng extractum is to the influence
of removal ovary rat fifth lumbar vertebra static parameter
(
*P＜0.05，
**P＜001vsSham；
△P＜0.05，
△△P＜0.01vs?OVX；
#P＜0.05，
##P＜0.01vs?LV)
Can know by table 2.6; Compare with the Sham group, %Tb.Ar, Tb.Th have reduced by 41.9% (P＜0.01), 38.0% (P＜0.01) respectively in the OVX group rat lumbar vertebra static parameter, and Tb.Sp has increased by 31.1% (P＜0.05); Explain that removal ovary has made the bone trabecula of rat lumbar vertebra significantly reduce; Microstructure is loose degenerates, and the bone resorption parameter has increase trend, but not statistically significant.Compare with the OVX group; The Tb.Sp of LV group 18.2% (P＜0.05) that descended; The Tb.Th of GF (H) has increased by 28.3% (P＜0.01); Other parameters have increase trend but not statistically significant, and visible livial and high dose Flos Ginseng extractum can improve the microstructural destruction of removal ovary rat lumbar vertebra girder, and removal ovary is caused the influence that lumbar vertebra is lost does not have significance.The %Tb.Ar of GF (L), Tb.Th have increased by 49.3% (P＜0.01), 36.5% (P＜0.01) respectively, Tb.Sp 19.1% (P＜0.05) that descended, and the lumbar vertebra that prompting GF (L) can effectively prevent removal ovary to cause is lost and microstructural destruction.
2.7 Flos Ginseng's extractum is to the influence of removal ovary rat fifth lumbar vertebra morphometry dynamic parameter
Flos Ginseng's extractum is seen table 2.7 to the influence of dynamic parameter %L.Pm (fluorescent perimeter percentage rate) in the removal ovary rat fifth lumbar vertebra osseous tissue morphometry, MAR (bone mineralising deposition) and three kinds of bone formation rate index BFR/BS, BFR/BV, BFR/TV.
Table 2.7 Flos Ginseng extractum is to the influence
of removal ovary rat fifth lumbar vertebra dynamic parameter
(
△△P＜0.01vs?OVX；
#P＜0.05，
##P＜0.01vs?LV)
Can know that by table 2.7 compare with the Sham group, the fifth lumbar vertebra dynamic parameter of OVX group rat has increase trend, but not statistically significant, the prompting removal ovary is to the bone formation and the not obviously effect of bone mineralising of rat lumbar vertebra.Compare with the OVX group, the MAR% of prevention group, BFR/BV reduce, but not statistically significant is explained bone formation and the bone mineralising not obviously influence of medicine to removal ovary rat lumbar vertebra.
2.8 Flos Ginseng's extractum is to the influence of removal ovary rat ulna physical index
Flos Ginseng's extractum is seen table 2.8 to the influence of removal ovary rat ulna weight in wet base (BWW), dry weight (BDW), weight in wet base/weight ratio (BWW/BW), dry weight/weight ratio (BDW/BW).
Table 2.8 Flos Ginseng extractum is to the influence
of removal ovary rat ulna physical index
(
**P＜001vs?Sham；
△P＜0.05，
△△P＜0.01vsOVX；
##P＜0.01vsLV)
Can be known by table 2.8, compare with the Sham group that BWW/BW, the BDW/BW of OVX group rat ulna reduce by 13.2%, 11.9% (P＜0.01) respectively, the prompting removal ovary can make ulna weight reduce.Compare with the OVX group, BWW/BW, the BDW/BW of LV group rat ulna increase by 23.9%, 24.3% (P＜0.01) respectively; BWW/BW, the BDW/BW of GF (L) group rat ulna increase by 8.7%, 8.1% (P＜0.05) respectively; BWW/BW, the BDW/BW of GF (H) group rat ulna increase by 13.0%, 13.5% (P＜0.01) respectively, explain that LV, GF can effectively prevent the reduction of rat ulna weight due to the removal ovary, and GF (H) effect are superior to GF (L).Compare with the LV group, BWW/BW, the BDW/BW of GF (L) and GF (H) obviously reduce, and explain that Flos Ginseng's extractum is to the too late livial of the improvement of the reduction of removal ovary rat ulna weight.
2.9 Flos Ginseng's extractum is to the influence of removal ovary rat ulna biochemical indicator
Flos Ginseng's extractum is seen table 2.9 to the influence of the Hyp of removal ovary rat ulna (bone hydroxyproline/backbone is heavy), Ca (calcium content of bone/backbone is heavy), P (bone phosphorus content/backbone's weight), Ca/Hyp (calcium/hydroxyproline content ratio).
Table 2.9 Flos Ginseng extractum is to the influence
of removal ovary rat ulna biochemical indicator
(
△P＜0.05，
△△P＜0.01vsOVX；
#P＜0.05，
##P＜0.01vsLV)
Result by table 2.9 is visible, compares with the Sham group, and OVX group rat femur calcium content reduces, but not statistically significant, Hyp content and Ca/Hyp ratio do not have significant change, and the prompting ovariectomized rats can not cause losing of ulna inanimate matter and organic matter after 12 weeks.Compare with the OVX group, LV group calcium content of bone and Ca/Hyp ratio obviously increase (P＜0.01); Hyp content and the Ca/Hyp ratio of GF (L) group rat obviously increase (P＜0.01); The Hyp content of GF (H) group rat obviously increases (P＜0.05), and the prompting livial can increase the content of removal ovary rat inanimate matter, and Flos Ginseng's extractum can increase the organic content of removal ovary rat.Compare with the LV group, the Hyp content of GF (L) group rat obviously increases (P＜0.01), and it is stronger than livial that prompting low dosage Flos Ginseng extractum improves the effect of the organic content of removal ovary rat.
2.10 Flos Ginseng's extractum is to the influence of removal ovary rat femur biomechanical property
Flos Ginseng's extractum is to the evaluation skeletal structure performance (elastic load, maximum load, breaking load) of removal ovary rat femur, and table 2.10 is seen in the influence of the external hardness of evaluating material (rigidity), toughness indexs such as (flexionals).
Table 4.10 Flos Ginseng extractum is to the influence of removal ovary rat femur biomechanics
(
*P＜0.05，
**P＜0.01vs?Sham；
△P＜0.05，
△△P＜0.01vsOVX；
##P＜0.01vsLV)
Can know by table 2.10, with Sham group relatively, elastic load, maximum load, the breaking load of OVX group rat femur descended respectively 19.7% (P＜0.01), 13.0% (P＜0.01), 10.6% (P＜0.05).The prompting removal ovary can make the biomechanical property of rat femur obviously weaken, and is prone to produce fracture.Compare with the OVX group, the biomechanics index of each group of prophylactic agent all has elastic load, maximum load, breaking load, stiffness coefficient and the flexional of obvious increase: LV group to increase by 65.9% (P＜0.01), 17.3% (P＜0.01), 16.0% (P＜0.01), 129.8% (P＜0.01), 129.8% (P＜0.01) respectively; The stiffness coefficient of Flos Ginseng's extractum femur of low, high dose increases by 139.7% (P＜0.01) and 139.7% (P＜0.01) respectively, and flexional has increased by 141.6% (P＜0.01) and 139.6% (P＜0.01) respectively; The elastic load of high dose increases by 22.4%.The biomechanical property of pointing out each prevention group effectively to prevent removal ovary to cause weakens.Compare with the LV group; Elastic load, maximum load, the breaking load of GF (L) group obviously reduce (P＜0.01); Elastic load, the breaking load of GF (H) group obviously reduce (P＜0.05), explain that Flos Ginseng's extractum is not so good as livial to the raising of removal ovary rat femur bone strength.
3. discuss
3.1 the bone amount of Flos Ginseng's extractum tibia epimere obviously increases, the index of reflection bone formation and bone mineralising descends
Compare with OVX group, the bone trabecula showed increased of Flos Ginseng's extractum (L) and Flos Ginseng's extractum (H) group, increase slightly, seriality is good; The Tb.N of Flos Ginseng's extractum (L) increases by 47.1% (P＜0.05), and the %Tb.Ar of Flos Ginseng's extractum (H), Tb.Th have increased by 64.8% (P＜0.05), 27.1% (P＜0.01) respectively; Though Tb.Sp descends, not statistically significant.Flos Ginseng's extractum that high and low dose is described all can effectively prevent bone loss and the microstructural destruction of removal ovary to the rat spongy bone.Bone resorption parameter Oc.N/mm has reduced by 65.4% (P＜0.05), explains that Flos Ginseng's extractum has remarkable inhibitory action to osteoclast.The dynamic indicator aspect, the %L.Pm of GF (L), MAR, BFR/BS, BFR/BV have reduced by 60.8%, 21.9%, 68.6%, 72.6% respectively; The %L.Pm of GF (H), MAR, BFR/BS, BFR/BV have reduced by 75.3%, 44.5%, 85.6%, 88.8% respectively.Explain that Flos Ginseng's extractum had both suppressed the bone resorption of removal ovary rat tibia epimere spongy bone, suppress its bone formation again, the effect that suppresses bone resorption is far longer than and suppresses osteoplastic effect, so the bone amount increases.
3.2 Flos Ginseng's extractum significantly increases the bone amount of removal ovary rat tibia stage casing cortical bone, medullary cavity reduces; The formation of perimyelis and mineralising are active to be reduced
With OVX group relatively, the %Ct.Ar of GF (L), GF (H) group increases by 4.6% (P＜0.01) and 4.3% (P＜0.01) respectively, %Ma.Ar descend respectively 14.8% (P＜0.01) and 14.0% (P＜0.01); Prompting Radix Ginseng flower concrete increases removal ovary rat cortical bone area, and medullary cavity is dwindled, and explains that the cortical bone bone amount that Flos Ginseng's extractum can effectively resist the removal ovary rat obviously reduces.Compare with the OVX group; The %E-L.Pm of GF (L), GF (H) group descend respectively 43.7% (P＜0.05) and 69.0% (P＜0.01); But other dynamic parameters do not have significant change; Explain that Flos Ginseng's extractum has resisted the bone formation of removal ovary rat tibia stage casing cortical bone inner membrance and the increase of mineralising, does not have obvious influence to periosteum.
3.3 Flos Ginseng's extractum obviously increases removal ovary rat lumbar vertebra amount, the index of reflection bone formation and bone mineralising changes not obvious
Compare with the OVX group, the Tb.Th of GF (H) has increased by 28.3% (P＜0.01), and other parameters have increase trend but not statistically significant; The %Tb.Ar of GF (L), Tb.Th have increased by 49.3% (P＜0.01), 36.5% (P＜0.01) respectively; Tb.Sp 19.1% (P＜0.05) that descended; The lumbar vertebra that prompting GF (L) can effectively prevent removal ovary to cause is lost and microstructural destruction; And GF (H) has only improved the tiny form of removal ovary rat lumbar vertebra girder, fails effectively to resist the bone amount of removal ovary rat lumbar vertebra and significantly loses.Compare with the OVX group, the MAR% of prevention group, BFR/BV reduce, but not statistically significant is explained bone formation and the bone mineralising not obviously influence of Flos Ginseng's extractum to removal ovary rat lumbar vertebra.
3.4 Flos Ginseng's extractum obviously improves the biomechanical property of removal ovary rat femur
That bone biomechanical is observed is low, the stiffness coefficient of Flos Ginseng's extractum femur of high dose increases by 139.7% (P＜0.01) and 139.7% (P＜0.01) respectively, and flexional has increased by 141.6% (P＜0.01) and 139.6% (P＜0.01) respectively; The elastic load of high dose increases by 22.4%.External hardness, toughness and bone strength that high dose Flos Ginseng extractum femur is described all obviously strengthen, and low dosage Flos Ginseng extractum obviously increases the external hardness and the toughness of removal ovary rat femur.
3.5 Flos Ginseng's extractum increases removal ovary rat ulna weight, the content of organic matter increases
BWW/BW, the BDW/BW of GF (L) group rat ulna increase by 8.7%, 8.1% (P＜0.05) respectively; BWW/BW, the BDW/BW of GF (H) group rat ulna increase by 13.0%, 13.5% (P＜0.01) respectively.Explain that LV, GF can effectively prevent the reduction of rat ulna weight due to the removal ovary, and GF (H) effect is superior to GF (L).Hyp content and the Ca/Hyp ratio of GF (L) group rat obviously increase (P＜0.01); The Hyp content of GF (H) group rat obviously increases (P＜0.05).Prompting Radix Ginseng flower concrete can increase the organic content of removal ovary rat.
4. conclusion
Flos Ginseng's extractum has tangible control people effect to the removal ovary osteoporosis rat, and its characteristics are:
1. the bone amount of Flos Ginseng's extractum tibia epimere obviously increases, and bone formation rate and bone resorption descend.
2. Flos Ginseng's extractum significantly increases the bone amount of removal ovary rat tibia stage casing cortical bone, and medullary cavity reduces; The formation of perimyelis and mineralising are active to be reduced.
3. Flos Ginseng's extractum obviously increases removal ovary rat lumbar vertebra amount, and the index of reflection bone formation and bone mineralising changes not obvious.
4. Flos Ginseng's extractum obviously improves the biomechanical property of removal ovary rat femur.
5. Flos Ginseng's extractum increases removal ovary rat ulna weight, and the content of organic matter increases.
Claims (7)
1. the Flos Ginseng is in the pharmaceutical preparation of preparation prevention of osteoporosis and the application of functional food.
2. the preparation with Flos Ginseng preparation as claimed in claim 1 is characterized in that said preparation is through following prepared: get the Flos Ginseng, pulverize, add 15 times of boiling water and soaked 30 minutes, filter, keep filtrating; Filtering residue adds 15 times of boiling water again and soaked 30 minutes, filters, and keeps filtrating; Filtering residue added 10 times of 60% soak with ethanol after 24 hours, and heating extraction 2 hours is filtered, and reclaims ethanol, must filtrate, and merges filtrating three times, concentrates, and drying is processed the preparation that can supply clinical practice by pharmaceutics technology.
3. the preparation with Flos Ginseng preparation as claimed in claim 1 is characterized in that said preparation is through following prepared: get the Flos Ginseng, pulverize, carry out micronization with the Baily pulverizing mill, add 15 times of boiling water and soaked 30 minutes, filter, keep and filtrate; Filtering residue adds 15 times of boiling water again and soaked 30 minutes, filters, and keeps filtrating; Filtering residue adds 10 times of 60% soak with ethanol 24 hours, after, heating extraction 2 hours is filtered, and reclaims ethanol, must filtrate, and merges filtrating three times, concentrates, and drying is processed the preparation that can supply clinical practice by pharmaceutics technology.
4. the preparation with Flos Ginseng preparation as claimed in claim 1 is characterized in that said preparation is through following prepared: get the Flos Ginseng, pulverize, carry out micronization with the Baily pulverizing mill, add 15 times of 20 ℃～25 ℃ distilled water immersions 30 minutes, filter, keep and filtrate; Filtering residue adds 15 times of 20 ℃～25 ℃ distilled water immersions 30 minutes again, filters, and keeps filtrating; Filtering residue adds 10 times of distilled water, transfers PH to 1 with hydrochloric acid; Reacting by heating 2 hours is put coldly, adds alkali and transfers PH to 7; Filter, must filtrate.Merge filtrating three times, concentrate, drying is processed the preparation that can supply clinical practice by pharmaceutics technology.
5. the preparation with Flos Ginseng preparation as claimed in claim 1, it is characterized in that said preparation is through following prepared: get the Flos Ginseng, pulverize, add 15 times of 70% soak with ethanol 24 hours, heating extraction 2 hours is filtered, and must filtrate.Filtering residue adds 40% soak with ethanol 24 hours again, and heating extraction 2 hours is filtered, and must filtrate.Merge filtrating twice, concentrate, drying is processed the preparation that can supply clinical practice by pharmaceutics technology.
6. the preparation with Flos Ginseng preparation as claimed in claim 1 is characterized in that said preparation is through following prepared: get the Flos Ginseng, pulverize; Carry out micronization with the Baily pulverizing mill, add 15 times of 70% soak with ethanol 24 hours, heating extraction 2 hours; Filter, must filtrate.Filtering residue adds 40% soak with ethanol 24 hours again, and heating extraction 2 hours is filtered, and must filtrate.Merge filtrating twice, concentrate, drying is processed the preparation that can supply clinical practice by pharmaceutics technology.
7. like claim 2,3,4,5,6 describedly process the preparation that can supply clinical practice with the Flos Ginseng by pharmaceutics technology, and these preparations are meant tablet, pill, electuary, granule, capsule, injection, drop pill, the preparation for external application to skin by Flos Ginseng's preparation.
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| CN106387551A (en) * | 2016-09-18 | 2017-02-15 | 天津北洋百川生物技术有限公司 | Ginseng flower bud and raspberry composite beverage and preparation technology thereof |
| CN108567140A (en) * | 2018-05-03 | 2018-09-25 | 璁哥究 | A kind of preparation method for the plant source calcium supplementing product being easily absorbed by the body |
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