CN102266415A - Application of compound traditional Chinese medicine to prevention and treatment of osteoporosis disease - Google Patents

Abstract

The invention discloses application of a compound traditional Chinese medicine to prevention and treatment of an osteoporosis disease, belonging to new application of the traditional Chinese medicine. The compound traditional Chinese medicine is prepared from the following Chinese medicinal herbs in percentage by weight: 15-35 percent of glossy privet fruit, 10-25 percent of Japanese thistle herb or root, 5-20 percent of largehead atractylodes rhizome, 5-20 percent of root of red-rooted salvia, 5-15 percent of coptis root, 5-15 percent of pseudo-ginseng root, 5-15 percent of eucommia and 5-15 percent of fingered citron. As proved by an experiment, the compound traditional Chinese medicine has remarkable treatment effects on the prevention and treatment of primary osteoporosis, secondary osteoporosis, osteoporosis caused by hyperlipidaemia or an osteoporosis disease caused by glucose metabolism syndrome, definite clinical curative effect and no remarkable side effect according to observation.

Description

The application of a kind of herbal mixture in preventing and treating the osteoporosis disease

Technical field

The present invention relates to field of medicaments, be specifically related to a kind of application of herbal mixture in preventing and treating the osteoporosis disease that contains Fructus Ligustri Lucidi.

Background technology

Osteoporosis (osteoporosis) is the commonly encountered diseases among the old people, frequently-occurring disease, and its cardinal symptom and harm have pain, height to shorten, fracture, respiratory function descend when bow-backed, serious.

Osteoporosis is one group of osteopathia that multiple reason causes, osseous tissue has normal calcification, and calcium salt and substrate are normal ratio, and the metabolic osteopathy that is reduced to characteristics with unit volume internal skeleton tissue mass becomes.The visible cortical bone of pathological anatomy is poor, and the sparse atrophy osteoid of bone trabecula layer is not thick.It is because the complicated result of factors such as heredity, hormone and nutrition under influencing each other.Calcium and vitamin D deficiency, hyperthyroidism, take glucocorticoid, smoking, excessive drinking and long-term bed etc. inadequately and all can cause osteoporosis.

But by its cause of disease osteoporosis dtex property sent out (constitutional) and Secondary cases two big classes.Essential divides juvenile form, adult, menopause, senile; Secondary cases is that medicine or trophic factor and other disease cause, such as endocrine regulation with excessively use corticosteroid and hepatorenal disease etc. all can cause secondary osteoporosis.

The mechanism of tracing it to its cause, gonadal hormone is the synthetic key factor of skeleton.Senile osteoporosis may be low with sex hormone level, the protein synthesis metabolism stimulate weaken and function of osteoblast goes down, sclerotin form reduce etc. relevant.Estrogen has the osteoclast activity of inhibition, reduces bone resorption, promotes effects such as osteoblast activity and the formation of promotion sclerotin, and the effect of antagonism hydrocortisone and thyroxin is arranged.Estrogen reduces after menopause, and bone resorption quickens and osteoporosis takes place gradually.Estrogen stimulates the l-a-hydroxylase to produce the effect of 1.25-(OH) 2-D3 in addition.Lack gonadal hormone after climacteric, 1-a one hydroxylase lowers the sensitivity that stimulations such as parathyroid hormone (PTH) low serum p generate, 1.25-(OH) the 2-D3 biosynthesis is low, also participate in osteoporosis taking place with advancing age, osteoblast is dead gradually, bone matrix all shows the most outstanding changing the performance, particularly osseous tissue that therefore senile osteoporosis is actually the organism aging process process in amount aspect matter.

The body hydrocortisone increases, because glucocorticoid is suppressed to the movable formation that influences bone matrix of osteocyte, suppresses the intestinal calcium absorption, increase the output protein synthesis inhibition simultaneously of urine calcium, decomposing increases, and causes negative calcium and negative nitrogen balance to make the osteosis obstacle, but mainly is that bone absorption increases.

At present, the medicine three major types of protect against osteoporosis: 1. bone resorption inhibitor, to reduce further losing of bone amount: estrogen, calcitonin, diphosphate, ipriflavone all belong to this class; 2. bone formation-promoter to increase the bone amount, comprises fluoride, vitamin K, parathyroid hormone, androgen, growth hormone etc.; 3. bone mineralising promoter promotes the bone calcific deposit, increases the bone amount, and this class medicine has vitamin D and calcium preparation.

Shortcomings such as at present, modern medicine control sclerotin is dredged disease gonadal hormone commonly used and fluoride etc., and existence costs an arm and a leg, and toxic and side effects is many, and late result is sure.

Chinese patent 200410051250.4 discloses a kind of medicine for the treatment of hyperlipemia, by Fructus Ligustri Lucidi, the Rhizoma Atractylodis Macrocephalae, the Cortex Eucommiae, Radix Notoginseng, Radix Cirsii Japonici, Radix Salviae Miltiorrhizae, Rhizoma Coptidis, Fructus Citri Sarcodactylis eight flavor Chinese medicines are formed, be through clinical nearly two more than ten years application experience side, be used for the treatment of hyperlipemia and arteriosclerosis, bran urine is sick, diseases such as metabolic syndrome, its clinical practice is existing for many years, but does not see to be useful on and prevent and/or treat the osteoporosis relevant disease and comprise constitutional, secondary osteoporosis, hyperlipemia or glycolipid metabolism syndrome and or the osteoporotic report that causes of medicine source property (as glucocorticoid).

Summary of the invention

The objective of the invention is to according to the deficiencies in the prior art, the application of a kind of herbal mixture in preventing and treating the osteoporosis disease is provided, described herbal mixture is the medicament of being made by the raw material of Chinese medicine of following percentage by weight:

Fructus Ligustri Lucidi 15 ~ 35%, Radix Cirsii Japonici 10 ~ 25%, the Rhizoma Atractylodis Macrocephalae 5 ~ 20%, Radix Salviae Miltiorrhizae 5 ~ 20%, Rhizoma Coptidis 5 ~ 15%, Radix Notoginseng 5 ~ 15%, the Cortex Eucommiae 5 ~ 15% and Fructus Citri Sarcodactylis 5 ~ 15%, called after VAOF.

Theory of Chinese medical science is thought " the kidney being the origin of congenital constitution ", " kidney producing bone marrow ", " it fills at bone ".Powerful weak and kidney essense prosperity and decline of bone growth and development is closely related, and then marrow biochemistry is active for the kidney essense abundance, and skeleton is nourished and strong strong; Then bone marrow biochemistry is passive for deficiency of kidney-essence, and skeleton loses foster and unable a little less than the flaccidity.Fundamentally recognizing suffers from a deficiency of the kidney is osteoporotic main pathogenesis.

The inventor has the effect of the osteoporosis of preventing and/or treating relevant disease by discovering Chinese patent 200410051250.4 disclosed a kind of medicines (being herbal mixture of the present invention) for the treatment of hyperlipemia.Described osteoporosis relevant disease comprise constitutional, secondary osteoporosis, hyperlipemia or glycolipid metabolism syndrome and or the osteoporosis that causes of medicine source property (as glucocorticoid), especially the osteoporosis that hyperlipemia, carbohydrate metabolism syndrome and glucocorticoid are caused has notable therapeutic effect

Show that through pharmacological research this herbal mixture can significantly improve the long-pending and tibia epimere of osteoporosis model animal middle tibia cortical bone surface of bone and organize bone trabecular bone amount, number and width, and significantly reduce its medullary cavity area and bone trabecular separating degree; All can significantly increase unit osteoblast number; Can significantly increase the bone amount of hyperlipemia rat cortical bone and spongy bone, improve bone structure; Also prevent the reduction of aged animal and human's sex hormone level etc., have effects such as the gonad function of enhancing.Based on above-mentioned pharmacological action, herbal mixture of the present invention can be used for prevention and treatment osteoporosis, treatment constitutional, secondary osteoporosis, hyperlipemia or glycolipid metabolism syndrome and or the osteoporosis clinical efficacy that causes of medicine source property (as glucocorticoid) definite, have effects such as the sexual organ development of enhancing function, and do not observe apparent side effect.Therefore, this herbal mixture can be used in preparation and prevents and treats the medicine of osteoporosis disease.

According to purposes of the present invention, wherein prevent and/or treat the medicine of osteoporosis relevant disease and/or the form that health product can be made for tablet, capsule, powder, pill, powder (injectable powder etc.), granule, crystal, solution, extractum, outstanding agent, soup, syrup, elixir, tea, oil, soft capsule, drop pill etc.

Compared with prior art, the present invention has following beneficial effect:

The present invention provides a kind of Chinese patent medicine new product with determined curative effect, cheapness for preventing and/or treating the osteoporosis relevant disease.The present invention confirms that through the modern Chinese medicine pharmacological research this herbal mixture has following function:

1) has and improve the long-pending and tibia epimere of osteoporosis model animal middle tibia cortical bone surface of bone and organize bone trabecular bone amount, number and width, and significantly reduce its medullary cavity area and bone trabecular separating degree; All can significantly increase unit osteoblast number; Can significantly increase the bone amount of hyperlipemia rat cortical bone and spongy bone, improve the effect of bony structures;

2) prevent the reduction etc. of aged animal and human's sex hormone level in addition, have effects such as the sexual organ development of enhancing function;

3) clinical research result shows that this herbal mixture has significant increase middle-aged and elderly people bone density, prevents the effects such as reduction of old man's sex hormone level that treatment osteoporosis relevant disease clinical efficacy is definite, and does not observe apparent side effect.

Description of drawings

Fig. 1. high fat diet and VAOF be to the morphologic influence of rat TX, scale 45 μ m, light field, amplification is 10 * 2.

Fig. 2. high fat diet and VAOF be to the morphologic influence of P of Rats TM, scale 5 μ m, light field, amplification is 10 * 2.

The specific embodiment

In order to understand essence of the present invention better, test in animal body with herbal mixture of the present invention (VAOF) below and human clinical trial's result further specifies the present invention, but protection scope of the present invention is not limited to this.

Herbal mixture (VAOF): Fructus Ligustri Lucidi 15 ~ 35%, Radix Cirsii Japonici 10 ~ 25%, the Rhizoma Atractylodis Macrocephalae 5 ~ 20%, Radix Salviae Miltiorrhizae 5 ~ 20%, Rhizoma Coptidis 5 ~ 15%, Radix Notoginseng 5 ~ 15%, the Cortex Eucommiae 5 ~ 15% and Fructus Citri Sarcodactylis 5 ~ 15%.

Embodiment 1 VAOF goes the experimentation of testis osteoporosis rat in control

It is one of osteoporotic important pathogenic factor of elderly men that androgen reduces.The osteoporosis model of suffering from a deficiency of the kidney that this research adopts 8 monthly age male rats to go testis to duplicate, VAOF with various dose prevents and treats, and compare research with positive drug tridin (Tridin), be intended to inquire into the therapeutic mechanism of this herbal mixture treatment male osteoporosis, for the further developmental research of VAOF provides scientific basis.

1. material

(1) medicine: VAOF, provide (Fructus Ligustri Lucidi 15, Radix Cirsii Japonici 10, the Rhizoma Atractylodis Macrocephalae 10 by the Chinese medicine academy of Guangdong Pharmaceutical University, Radix Salviae Miltiorrhizae 10, Rhizoma Coptidis 5, Radix Notoginseng 5, the Cortex Eucommiae 10 and Fructus Citri Sarcodactylis 10), lot number: 100502, positive drug tridin (Tridin) provides lot number by Italian Rotta pharmaceutical factory: 2010024, and this medicine is used to treat the loaded European various countries of osteoporosis pharmacopeia.

(2) animal: select 60 of 8 monthly age male SD rats for use, body weight 280 ~ 300g, Guangdong Medical Lab Animal Center provides, the quality certification number: 05A018.

2. experimental technique

(1) modeling method: 60 laboratory animals are divided into 6 groups at random, 10 every group, are respectively normal control group, model group, the high, medium and low dosage group of VAOF and tridin group.Each organize rat all with 3% pentobarbital sodium by 1ml/kg body weight intraperitoneal injection of anesthesia after, the sham-operation of normal control group row, all the other respectively organize rat all from the capable complete bilateral testes enucleation of veutro approach.

(2) raising and route of administration: postoperative beginning in the 3rd day administration, normal control group, model group are irritated stomach with distilled water 6ml/kg every day, the tridin group gives tridin 60mg/kg and irritates stomach, be made into suspension before the use, the high, medium and low dosage group of VAOF gives high, medium and low dosage VAOF every day respectively and irritates stomach.All rats are all under equal conditions fed, and freely take the photograph water, control appetite.

3. observation index

(1) bone density detects: each is organized rat and used the U.S. QDR-2000 of HOLOGIG company type dual intensity X line absorption instrument (coefficient of variation (CV)＜1.0%) scanning whole body bone mineral content and bone density in preceding 2 days under anesthesia in putting to death, and gets two sensitive spots of right femur and fl lower end respectively.

(2) mensuration of serum biochemistry index: before the modeling and put to death that to get the centrifugal back of blood before the animal respectively standby, radio immunoassay is measured testosterone (T), IL-6 and BGP content.IL-6, BGP test kit are put hands-free confession by PLA General Hospital Science and Technology Development Center, and T is put the box of being excused from an examination and provided by Tianjin DPC company, and gauge is a BECKMAN DP5500 type r enumerator.

Statistical procedures: all data all adopt mean+standard deviation (x+SD) expression, analyze with the SPSS statistical software, relatively use variance analysis between group, and significant difference is checked with q.

4. result

(1) take each dosage VAOF and tridin the control go testis rat body and left and right femur density (BMD) and bone calcium mineral content (BMC) all be higher than model group, difference has significance (P＜0.05 ~ 0.01).

The influence that table 1:VAOF changes the castrated rats BMD (n=10 of x ± SD)

Group	Dosage (g/kg)	Right femur	Fl
				The normal control group	——	0.168±0.017	0.189±0.016
Model group	——	0.135±0.009 **	0.146±0.011 **
				Model+tridin	0.06	0.168±0.019 ▲▲	0.177±0.018 ▲▲
Model+VAOF is low	1	0.161±0.018 ▲▲	0.171±0.014 ▲▲
				Among model+VAOF	2	0.169±0.016 ▲▲	0.177±0.016 ▲▲
Model+VAOF height	4	0.170±0.023 ▲▲	0.183±0.018 ▲▲

Annotate: compare with normal group, ^*P＜0.05, ^*P＜0.01; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01.

The influence that table 2:VAOF changes the castrated rats BMC (n=10 of X ± SD)

Group	Dosage (g/kg)	Right femur	Fl
				The normal control group	——	0.069±0.007	0.075±0.008
Model group	——	0.053±0.004 **	0.0561±0.006 **
				Model+tridin	0.06	0.067±0.009 ▲▲	0.071±0.007 *
Model+VAOF is low	1	0.063±0.009 ▲▲	0.068±0.009 ▲▲
				Among model+VAOF	2	0.066±0.008 ▲▲	0.069±0.008 ▲▲
Model+VAOF height	4	0.068±0.008 ▲▲	0.071±0.009 ▲▲

Annotate: compare with normal group, ^*P＜0.05, ^*P＜0.01; Compare with model group, ^{▲ ▲}P＜0.05, ^{▲ ▲}P＜0.01.

(2) remove testosterone (T) content there was no significant difference before the testis (P〉0.05) between each group, went behind the testis 4 months, T is apparently higher than model group for the high, medium and low dosage group of VAOF, and difference has significance (P＜0.05).

Table 3:VAOF is to the influence of removing testis rat TESTO (T) content (n=10 of X ± SD)

Group	Dosage (g/kg)	Before the art	Postoperative 4 months
				The normal control group	——	3.41±0.61	3.73±0.96 **
Model group	——	3.39±0.89	0.18±0.13
				Model+tridin	0.06	3.20±1.12	0.28±0.19
Model+VAOF is low	0.75	3.10±0.92	0.39±0.21 ▲
				Among model+VAOF	1.5	3.43±0.69	0.48±0.23 ▲▲
Model+VAOF height	4	3.32±0.85	0.79±0.45 ▲▲

Annotate: compare with normal group, ^*P＜0.05, ^*P＜0.01; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01.

The preceding serum BGP content indifference of operation between (3) each is organized (P〉0.05), the postoperative administration is after 4 months, model group is apparently higher than normal group (P＜0.01), and three groups and the tridin of VAOF control then are starkly lower than model group (P＜0.05 ~ P＜0.01).

Example 2 VAOF are to the osteoporotic preventive and therapeutic effect of ovariectomized rats

1. material

(1) medicine: VAOF, provide (Fructus Ligustri Lucidi 15, Radix Cirsii Japonici 10, the Rhizoma Atractylodis Macrocephalae 10 by the Chinese medicine academy of Guangdong Pharmaceutical University, Radix Salviae Miltiorrhizae 10, Rhizoma Coptidis 5, Radix Notoginseng 5, the Cortex Eucommiae 10 and Fructus Citri Sarcodactylis 10), lot number: 100502, positive drug calcareacarbonica sheet, White Cloud Mountain, Guangzhou brilliance pharmacy joint stock company limited, lot number: T90016; E ₂Detection kit, Kemei Dongya Biological Technology Co., Ltd., Beijing provides, lot number: 110325.

(2) animal: selected for 32 ages in week for use, 72 of cleaning level, SD female rats, body weight (390 ± 20g), by Guangdong Province's experiment thing center laboratory animal feed formula, giving normal diet feeds, with reference to AIN-93M rodent feed formula, with the composition that derives from Semen sojae atricolor in the suitable substitution of materials feedstuff, to avoid of the interference of phytohormone such as soybean isoflavone to experimental result.Freely take the photograph water, throw light on round the clock that the light and shade time of replacing is 12 hours, adaptability was fed 7 days.

(3) instrument: Japanese NORLAND dual intensity bone density meter, coefficient of variation CV＜0.1%, scanning times 2 times.Encircling the SN-6958B of a factory type wisdom Shanghai day puts and exempts from the r measuring instrument, CV＜1.19%.

2. method

Modeling method: rat is through the pentobarbital sodium solution anesthesia of lumbar injection 30mg/kgBW, the dried ventrimeson in fixing back, abdomen position is apart from vaginal orifice 3 ~ 4cm unhairing, use iodine tincture wiping skin and alcohol disinfecting respectively, skin and the about 2 ~ 3cm of abdominal muscle are cut in dried slightly back, the visible white bag fat in the otch visual field, after pushing fat deposit aside and finding the uterus, gently a side cornua uteri is pulled out the ovary that is as seen wrapped up at its end by liparitosis.Fractionation of fatty group just can see pink ovary, clamps ovary with mosquito forceps, then fallopian tube under the ovary (comprising fat) is used the silk thread ligation, wipes out ovary (check and whether wipe out fully), takes advantage of a situation the uterus is sent back to again in the abdominal cavity, wipes out other side ovary with method.After abdominal muscle and skin layering are sewed up, sterilization once more.After hindlimb muscle is injected 20,000 U Aminomycin.Sham-operated control group (10) is opened and is sewed up after abdomen is wiped out a fritter fat.Postoperative 5 days, animal begins the administration of dividing into groups after recovering, and the grouping situation sees Table.Sham-operated control group and oophorectomize group give normal saline 10ml/kg, and spay (OVX)+Western medicine group (calcium carbonate) gives calcareacarbonica sheet 0.2g/kg; Spay+low dosage VAOF group gives VAOF0.135g/kg(and is equivalent to 5 times of clinical people's consumption); Spay+middle dosage VAOF group gives VAOF 0.27g/kg (be equivalent to clinical people's consumption 10 times); Spay+high dose VAOF group gives VAOF 0.54g/kg (be equivalent to clinical people's consumption 20 times).Every day gastric infusion once, all administrations 6 days, altogether 16 weeks of administration.Duration of test continues to feed and drinking-water according to normal diet.

3. observation index

(1) mensuration of index such as serum biochemistry: 16 week of modeling the back femoral artery get blood, blood sample this treated after, separation of serum, each group 10 samples is at random carried out serum measured by radioimmunoassay estradiol (E ₂), interleukin-6 (IL-6); Complete taking-up right side femur is peeled off totally, claims weight in wet base; Separating uterus is weighed; Calculate the uterus index and the femur index of every 100g body weight.

(2) bone density: with the right side femur that takes out with Japanese NORLAND dual intensity bone density meter, scanning survey bone density and bone mineral content.

4. result

(1) changes of bone mineral density

Table 4 as seen, removal ovary is after 16 weeks, right femur density of oophorectomize group rat and bone mineral density all are starkly lower than Sham-operated control group, difference has significance (P＜0.01), shows the modeling success.Compare with the oophorectomize group, dosage group and heavy dose also can obviously improve OVX rat femur BMC content (P＜0.05 ~ P＜0.01) among the VAOF, the heavy dose of group of content (P＜0.05) that can obviously improve OVX rat femur BMD.

Table 4:VAOF to the influence of female OVX rat femur BMD, BMC ( ± s)

Group	n	Dosage (g/kg)	Right femur	Right femur
								BMD（g/cm 2）	BMC(g)
Sham operated rats	10	——	0.141±0.005 **	0.436±0.031 **
					The oophorectomize group	10	——	0.131±0.006	0.382±0.030
Spay+calcareacarbonica sheet	10	0.2	0.133±0.006	0.391±0.029
					Spay+VAOF is low	10	0.38	0.133±0.005	0.392±0.038
Among spay+VAOF	10	0.75	0.135±0.008	0.416±0.039 *
					Spay+VAOF height	10	1.5	0.139±0.009 *	0.434±0.041 **

Compare with model group: ^*P＜0.05, ^*P＜0.01.

(2) serum E ₂Variation with IL-6

Table 5 as seen, removal ovary is after 16 weeks, oophorectomize group rat blood serum E ₂Content significantly is lower than Sham-operated control group, and difference has significance (P＜0.05), shows the modeling success.Dosage group and high dose group E in the Chinese medicine ₂Content is apparently higher than model group, and difference has significance (P＜0.05-P＜0.01).

Table 5:VAOF is to female OVX rat E ₂With the influence of IL-6 (

± s)

Group	n	IL-6（pg/ml）	E 2(pmol/L)
				Sham-operated control group	10	81.8±34.6	141.6±34.8
The oophorectomize group	10	63.8±31.5	53.8±16.8
				Spay+calcareacarbonica sheet	10	66.2±21.9	71.8±21.8
Spay+VAOF is low	10	67.3±24.1	73.1±51.3
				Among spay+VAOF	10	69.2±25.1	86.9±33.8 *
Spay+VAOF height	10	77.5±43.9	111.3±40.3 *

Compare with model group: ^*P＜0.05, ^*P＜0.01.

(3) uterus exponential sum femur index

As seen table 6 compares with sham operated rats, and oophorectomize group uterus index and femur index obviously reduce, and have statistical significance (P＜0.05), prompting modeling success.Compare with the oophorectomize group, high dose group uterus index obviously increases (P＜0.05); High, medium and low three dosage group femur indexes all obviously increase (P＜0.05).

The table 6:VAOF to the exponential influence of female ovariectomized rat uterus exponential sum femur (

± s)

Group	n	Uterus index (g/100g)	Femur index (g/100g)
				Sham-operated control group	10	0.334±0.159 *	0.353±0.019 *
The oophorectomize group	10	0.037±0.011	0.282±0.022
				Spay+calcareacarbonica sheet	10	0.059±0.046	0.313±0.021 *
Spay+VAOF is low	10	0.053±0.059	0.310±0.023 *
				Among spay+VAOF	10	0.068±0.015	0.315±0.035 *
Spay+VAOF height	10	0.113±0.101 *	0.323±0.049 *

Compare with model group: ^*P＜0.05, ^*P＜0.01.

Discuss: above-mentioned experiment shows, compares with model group, and the present invention can obviously suppress rat blood serum E ₂Level decline, the right femur weight in wet base of inhibition descend and the exponential decline in uterus, show that the present invention has the reduction of prevention climacteric estrogen level and the atrophy of bone density improving function and the preventative organ of energy, sexual function improving.

Influence to the sclerotin of the unusual rat model of lipid metabolism

1. experiment material

(1) main medicine: VAOF, hyperlipidemia medicine VAOF: the Chinese medicine academy of Guangdong Pharmaceutical University provides (Fructus Ligustri Lucidi 15, Radix Cirsii Japonici 10, the Rhizoma Atractylodis Macrocephalae 10, Radix Salviae Miltiorrhizae 10, Rhizoma Coptidis 5, Radix Notoginseng 5, the Cortex Eucommiae 10 and Fructus Citri Sarcodactylis 10); Lot number: 100502.XUEZHIKANG JIAONANG: Beijing big dimension letter bio tech ltd (lot number: 20091116).

(2) main agents and instrument see the following form

Table 7 main agents

Reagent	Producer	Lot number
			NaTDC	Chemical Reagent Co., Ltd., Sinopharm Group	F20070521
Propylthiouracil	Guangdong Huanan Pharmaceutical Co., Ltd	070501
			Cholesterol	Tianjin Da Mao chemical reagent factory	20070518
Propylene glycol	Guangzhou Chemical Reagent Factory	20051203-1
			Tween 80	Shen, Shanghai space medication chemistry company limited	061201
Methyl methacrylate	1Guanghua Chemical Plant Co., Ltd., Guangdong	20080407
			Dibutyl phthalate	Tianjin big trade chemical reagent factory	20080829
Benzoyl peroxide	Tianjin good fortune chemical reagent in morning factory	20071102
			Hematoxylin	Amresco.USA	——
Poncean	Sigma Chemical Co.USA	——
			Fuchsin acid	Sigma Chemical Co.USA	——
Phosphotungstic acid	Sigma Chemical Co.USA	——
			Oranage G	Sigma Chemical Co.USA	——
Light green	Sigma Chemical Co.USA	——
			Ferric chloride	Sigma Chemical Co.USA	——
2-Methyloxethyl acetate	Sigma Chemical Co.USA	13110MD
			Glacial acetic acid	The Long Huagongshijichang of Chengdu section	20080812
Dehydrated alcohol	1Guanghua Chemical Plant Co., Ltd., Guangdong	20090421
			Anhydrous calcium chloride	1Guanghua Chemical Plant Co., Ltd., Guangdong	20081016
Dimethylbenzene	Tianjin good fortune chemical reagent in morning factory	20080902
			Neutral gum	New chemical plant is stood in Guangzhou	20081202
Yihong	Amresco	0109

8 key instrument equipment

Instrument	Producer
		T1000 type electronic balance	The two outstanding test instrunment of Changzhou factory
YM-III type Gypsum Fibrosum polisher	Northwest medical apparatus and instruments Group Co.,Ltd
		RM2500 type microtome	Leica Co.Germany
SP1600 type veneer sawing machine	Leica Co.Germany
		JB-2 type constant temperature blender with magnetic force	New river rising in Ningxia and flowing into central Shaanxi subsidiary factory of Shanghai thunder magnetic instrument plant
DHG-9145A type electric heating constant temperature air dry oven	Shanghai one permanent Science and Technology Ltd.
		Bioquant-Osteo bone morphological images analytical system	BIOQUANT USA

(3) laboratory animal: rat SPF level Spague Dawla(SD) is provided the quality certification: 2007A003 by Guangdong Province's Experimental Animal Center.

Experimental technique

(1) animal model is set up and administration

Modeling is with the preparation of lipomul: the lipomul preparation method: get 50g Adeps Sus domestica, be heated to and make its dissolving about 100 ℃, add the 10g cholesterol, be stirred to dissolving fully.Add 1g propylthiouracil and 25mL Tween 80 again, fully stir.Add 30mL distilled water and 20mL1 in another beaker, the 2-propylene glycol is heated to about 60 ℃, adds the 2g NaTDC, fully stirs up to dissolving.With two kinds of abundant mixings of solution, make lipomul.

Grouping and administration: under the experimental situation, in the 1st week, the normal raising divided normal group, lipomul group at random.The 2nd week, normal group: normal diet, freely drink water.The lipomul group: normal diet, freely to drink water, every morning is pressed 10mL/kg perfusion lipomul; After perfusion last day (the 14th day), lipomul compositional model group, positive drug group (Xuezhikang group), the high, medium and low dosage group of VAOF.Beginning (i.e. the 3rd week) in the 15th day, except that normal group, other are respectively organized every morning and continue the perfusion lipomul, irritate stomach after 4 hours respectively and give relative medicine.Xuezhikang group: XUEZHIKANG JIAONANG 0.25g/kg, the high, medium and low dosage group of VAOF respectively: 6,3,1.5g/kg.6 weeks of successive administration.Put to death rat, dissect and take out tibia, separate muscle, connective tissue attached thereto, tibia places 10% formalin conventional fixing, is replaced by 70% alcoholic solution behind 24 ~ 48h, and 4 ℃ of preservations are stand-by.

Table 9 grouping and administration situation

Group name	Label	Used medicine	Consumption
				Normal group	Ctrl	——	——
Model group	Mod	High lipoprotein emulsion	10mL/kg
				The Xuezhikang group	Xzk	High lipoprotein emulsion+XUEZHIKANG JIAONANG	10mL/kg+0.25g/kg
Low medicament group	VAOF-L	High lipoprotein emulsion+VAOF	10mL/kg+0.75 g/kg
				Middle medicament group	VAOF-M	High lipoprotein emulsion+VAOF	10mL/kg+1.5 g/kg
High medicament group	VAOF-H	High lipoprotein emulsion+VAOF	10mL/kg+3 g/kg

(2) osseous tissue morphometry method

Undecalcified osseous tissue sheet preparation: the embedding TX(shaft of tibia tuberosity of carrying out the undecalcified bone specimen of bone sample) and the PTM(proximal tibia) embedded block, cut on Leica RM2500 microtome, cut out the thin slice of 5 μ m, roasting sheet, be fixed on the cleaning processing of dewatering, dye on the microscope slide, mounting, be prepared into undecalcified osseous tissue sheet.Masson-Goldner Trichrome dyeing, after the dyeing, sclerotin is green, and bone marrow is red, and microscopically is observed bone trabecula quantity and its surperficial osteoblast and osteoclast.

：

1. the measuring range of measuring range: TX is the medullary cavity and the cortical bone of its transverse section; The measuring range of PTM to avoid elementary osteogenesis district, is down drawn 3 ㎜ from 1 ㎜ for 1 ㎜ that down draws from growth plate again.

2. the bone morphometry is measured and calculating parameter

This is tested all parameters and adopts the name of general international standard osseous tissue norphometry technics, directly measures with image analysis system.Chinese and English title, symbol and the unit of measurement parameter are shown in table 10 and table 11.But these parameters can't be used to analyze bone amount, bone structure, bone formation and bone resorption etc., the parameter that directly records are calculated by international formula, just can be used for analyzing.The Chinese of calculating parameter, symbol and computing formula are like shown in table 12 and the table 13.

Table 10: cortical bone measurement parameter

Chinese	English name	Symbol	Unit
				Proximal tibia	Proximal tibia	PTM
The shaft of tibia tuberosity	Tibia shaft	TX
				The osseous tissue gross area	Total tissue area	T.Ar	mm 2
The bone marrow gross area	Marrow area	Ma.Ar	mm 2
				The periosteum girth	Periosteal perimeter	P-Pm	mm
The perimyelis girth	Endocortical perimeter	E-Pm	mm
				Thickness of cortex of bone	Cortical thickness	Ct.Th	mm

Table 11 spongy bone measurement parameter

Chinese	English name	Symbol	Unit
				The osseous tissue area	Tissue area	T.Ar	mm 2
The bone trabecula area	Trabecular bone area	Tb.Ar	mm 2
				The bone trabecula girth	Trabecular perimeter	Tb.Pm	mm
The adherent girth of osteoclast	osteoclast perimeter	Oc.Pm	mm
				The adherent girth of osteoblast	osteoblast perimeter	Ob.Pm	mm
Amount of osteoclast	osteoclast number	N.Oc	#
				Osteoblast quantity	osteoblast number	N.Ob	#

Table 12 cortical bone calculating parameter and computing formula

Chinese	Symbol	Unit	Formula
				The cortical bone area percent	%Ct.Ar	%	(T.Ar- Ma.Ar)/T.Ar × 100
The medullary cavity area percent	% Ma.Ar	%	Ma.Ar/ T.Ar × 100

Table 13 spongy bone calculating parameter Tab5 Calculations of cancellous bone(PTM)

Chinese	Symbol	Unit	Formula
				The bone trabecula area percent	%Tb.Ar	%	Tb.Ar/T.Ar×100%
The bone trabecula width	Tb.Th	μm	（2000/1.199）×(Tb.Ar/Tb.Pm)
				Bone trabecula quantity	Tb.N	#/㎜	（1.199/2）×(Tb.Pm/T.Ar)
The bone trabecula separating degree	Tb.Sp	μm	（2000/1.199）×(Tb.Ar-T.Ar)/Tb.Pm
				Unit bone trabecula girth osteoblast number	Ob.N	#/㎜	Ob.N/Tb.Pm
Unit bone trabecula girth osteoclast number	Oc.N	#/㎜	Oc.N/Tb.Pm

3. the meaning of calculating parameter

(1) TX parameter

A. cortex area percent (%Ct.Ar): reflection cortical bone shared ratio in the total osseous tissue area of TX bone cross section;

B. medullary cavity area percent (%Ma.Ar): reflection medullary cavity shared ratio in the total osseous tissue area of TX bone cross section.

(2) PTM parameter

A. (%Tb.Ar, %): phalanges girder area accounts for the percentage rate of osseous tissue area to the bone trabecula area percent, and the bone amount what reflect.This index is to estimate the most important index of bone quantitative changeization.

B. bone trabecula width (Tb.Th): be used to describe the bone trabecula morphosis, explain the variation of bone amount.Under the certain situation of quantity, width is thick more, and the bone amount is many more.

C. bone trabecula quantity (Tb.N): be used to describe the bone trabecula morphosis, explain the variation of bone amount.Under the certain situation of width, quantity is many more, and the bone amount is many more.

D. bone trabecula separating degree (Tb.Sp): the average distance between the phalanges girder is used for describing bone trabecular morphosis.Separating degree is big more, and the distance between the bone trabecula is just big more, and bone is just loose more.

E. osteoblast number (Ob.N): the osteoblast number on the unit bone trabecula girth, reflect the bone formation situation relevant with osteoblast.

F. osteoclast number (Oc.N): the osteoclast number on the unit bone trabecula girth, reflect the bone resorption situation relevant with osteoclast.

3. result

(1) high fat diet and VAOF are to the influence of rat TX morphometry

As shown in Table 14: there is not significant difference in each group in gross area index.

As shown in Table 14: compare with the Ctrl group, Mod group cortical bone area percent obviously reduces (P＜0.05), and the medullary cavity area percent obviously increases (P＜0.05).

Compare with the Mod group, VAOF-L, VAOF-M group and VAOF-H group rat cortical bone area percent all obviously raise (P＜0.05), and result and Xzk group are similar.

Fig. 1 is seen in section photo as a result.

Table 14 high fat diet and VAOF are to the influence (n=10) of rat TX morphometry static parameter

Group Group	Osseous tissue gross area T.Ar (mm 2)	Cortical bone area percent Ct.Ar (%)	Medullary cavity area percent Ma.Ar (%)
				Ctrl	1.26±0.22	75.72±3.12	24.28±3.12
Mod	1.26±0.09	73.68±2.70 *	26.32±2.70 *
				Xzk	1.27±0.09	76.48±2.16 #	23.52±2.16 #
VAOF-L	1.22±0.09	76.87±2.26 #	23.13±2.26 #
				VAOF-M	1.21±0.08	77.08±2.66 #	22.92±2.66 #
VAOF-H	1.20±0.07	76.84±2.72 #	23.16±2.72 #

^*P<0.05 VS Ctrl； ^#P<0.05 VS Mod。

(2) high fat diet and VAOF are to the influence of P of Rats TM morphometry

As shown in Table 15: compare with the Ctrl group, bone trabecular bone amount and the width of Mod group rat significantly reduce (P＜0.01), and the bone trabecula number obviously reduces (P＜0.05), and separating degree obviously increases (P＜0.05); Compare with the Mod group: bone trabecular bone amount and the width increase of VAOF-H group and two medicine group rats of VAOF-M group all reach utmost point significant level (P＜0.01), and the result is identical with the Xzk group; In the bone trabecula number index, VAOF-H group increases and reaches significant level (P＜0.01), and the result organizes near Xzk, and VAOF-M in this index a little less than preceding two groups, but also obviously increase (P＜0.05); The VAOF-H group, VAOF-M group and Xzk group all obviously reduce bone trabecula separating degree (VAOF-H, M:P＜0.05; Xzk:P＜0.01).

Table 15 high fat diet and VAOF are to the influence (n=10) of P of Rats TM morphometry static parameter

Group Group	Bone trabecula area percentage %Tb.Ar (%)	Bone trabecula width Tb.Th (μ m)	Bone trabecula number Tb.N (#/mm)	Separating degree Tb.Sp (μ m)
					Ctrl	22.93±6.53	72.89±12.31	3.14±0.74	259.52±78.69
Mod	13.25±3.11 **	54.56±5.36 **	2.41±0.37 *	371.44±84.11 *
					Xzk	21.39±2.13 ##	62.01±7.80 #	3.47±0.20 ##	227.42±11.61 ##
VAOF-L	12.53±4.10 △△	52.47±2.43	2.10±0.73 △△	468.81±88.01 △△
					VAOF-M	19.14±3.93 ##	62.74±8.74 #	3.05±0.50 #	273.60±62.67 #
VAOF-H	20.40±3.13 ##	63.06±4.65 ##	3.23±0.36 ##	250.48±39.24 #

Contrast with normal group (Ctrl) ^*P＜0.05, ^*P＜0.01; With model group (Mod) contrast, ^#P＜0.05, ^##P＜0.01; Compare with the Xzk group ^△P＜0.05, ^{△ △}P＜0.01.

As shown in Table 16: compare with the Ctrl group, Mod group rat unit bone trabecula girth osteoblast digital display work reduces (P＜0.01); Compare with the Mod group, the middle and high dosage of VAOF group unit bone trabecula girth osteoblast digital display work increases (P＜0.01), and the result is consistent with the Xzk group; Each unit's of group bone trabecula girth osteoclast number does not have meaning difference.

The section experimental result is seen Fig. 2.

Table 16 high fat diet and VAOF are to the influence of P of Rats TM morphometry cell parameters

Group Group	The adherent girth of osteoblast (mm)	The adherent girth of osteoclast (mm)	The unit osteoblast is counted Ob.N (#/mm)	The unit osteoclast is counted Oc.N (#/mm)
					Ctrl	0.635±0.118 **	0.157±0.047	30.83±4.92	5.69±0.71
Mod	0.329±0.067	0.146±0.044	15.19±1.93 **	5.65±1.05
					Xzk	0.710±0.203 ##	0.160±0.033	24.20±2.47 ##	4.78±1.33
VAOF-L	0.248±0.091	0.107±0.025	14.87±3.25	5.08±1.81
					VAOF-M	0.555±0.065 ##	0.178±0.027	27.64±3.54 ##	6.38±1.29
VAOF-H	0.506±0.095 ##	0.178±0.044	22.84±4.56 ##	5.70±1.33

^*P<0.05 VS Ctrl； ^**P<0.01 VS Ctrl； ^#P<0.05 Vs Mod； ^##P<0.01 Vs Mod;

^△P<0.05 VS Xzk; ^△△P<0.01 VS Xzk。

Embodiment 4: VAOF brings out the influence of osteoporosis rat to hydrocortisone

It is one of osteoporotic important pathogenic factor of medicine source property that cortex alcohols medicine excessively uses.The bull rat is adopted in this research, the medicine source property osteoporosis model that the cortex alcohols medicine that subcutaneous injection hydrocortisone (36mg/kg) duplicates excessively brings out, VAOF with various dose prevents and treats, and compare research with tridin (Tridin), be intended to inquire into loyal art and transfer fat side's treatment cortex alcohols medicine excessively to use the medication of induced osteoporosis disease to imitate, transfer the further developmental research of fat side for loyal art the foundation of science is provided.

1. materials and methods

(1) medicine: VAOF provides (Fructus Ligustri Lucidi 15, Radix Cirsii Japonici 10, the Rhizoma Atractylodis Macrocephalae 10, Radix Salviae Miltiorrhizae 10, Rhizoma Coptidis 6, Radix Notoginseng 6, the Cortex Eucommiae 10 and Fructus Citri Sarcodactylis 10), lot number: 100503 by the Chinese medicine academy of Guangdong Pharmaceutical University; Positive drug tridin (Tridin) provides lot number by Italian Rotta pharmaceutical factory: 2010024, and this medicine is used to treat the loaded European various countries of osteoporosis pharmacopeia.

(2) animal: select 60 of adult male SD rats for use, body weight 280 ~ 300g, Guangdong Medical Lab Animal Center provides, the quality certification number: 09A018.

2. experimental technique

Be divided into 6 groups at random after 60 rats numbering weighed, every group 10, get wherein one group of subcutaneous injection normal saline as normal control group, one group of subcutaneous injection hydrocortisone (20mg/kg) as model group, one group of subcutaneous injection tridin (0.05g/kg) as the positive drug matched group, three groups of subcutaneous injection hydrocortisone (20mg/kg) are irritated the basic, normal, high dosage of stomach VAOF (0.75,1.5,3.0g/kg) more respectively in addition.Each group difference gastric infusion 1 time/day, normal control group and hydrocortisone group give the equal-volume normal saline, successive administration 30 days.All rats are all under equal conditions fed, and freely take the photograph water, control appetite.

3. observation index

Bone density detects: each is organized rat and used the U.S. QDR-2000 of HOLOGIG company type dual intensity X line absorption instrument (coefficient of variation (CV)＜1.0%) scanning whole body calcium content of bone and bone density in preceding 2 days under anesthesia in putting to death, and gets two sensitive spots of right femur and fl lower end respectively.

4. statistical procedures

All data all adopt mean+standard deviation (x+SD) expression, analyze with the SPSS statistical software, relatively use variance analysis between group, and significant difference is checked with q.

5. experimental result

Left and right femur density of hydrocortisone induced osteoporosis rat (BMD) and the bone calcium mineral content (BMC) of taking each dosage VAOF control all are higher than model group, and difference has significance (p＜0.05 ~ 0.01).

The influence that table 17 VAOF changes the hydrocortisone induced osteoporosis rat BMD (n=10 of x ± SD)

Group	Dosage (g/kg)	Right femur	Fl
				The normal control group	——	0.181±0.018	0.186±0.016
Model group	——	0.136±0.009 **	0.146±0.009 **
				Model+tridin	0.05	0.173±0.017 ▲▲	0.169±0.014 ▲▲
Model+VAOF is low	0.75	0.162±0.017 ▲▲	0.167±0.015 ▲▲
				Among model+VAOF	1.50	0.169±0.016 ▲▲	0.175±0.018 ▲▲
Model+VAOF height	3.0	0.175±0.025 ▲▲	0.181±0.019 ▲▲

Annotate: compare with normal group, ^*P＜0.05, ^*P＜0.01; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01.

The influence that table 18 VAOF changes the hydrocortisone induced osteoporosis rat BMC (n=10 of X ± SD)

Group	Dosage (g/kg)	Right femur	Fl
				The normal control group	——	0.061±0.007	0.075±0.008
Model group	——	0.049±0.006 **	0.057±0.006 **
				Model+tridin	0.05	0.058±0.009 ▲▲	0.069±0.014 ▲
Model+VAOF is low	0.75	0.056±0.008 ▲	0.067±0.008 ▲▲
				Among model+VAOF	1.5	0.058±0.008 ▲▲	0.069±0.007 ▲▲
Model+VAOF height	3.0	0.061±0.008 ▲▲	0.073±0.009 ▲▲

Annotate: compare with normal group, ^*P＜0.05, ^*P＜0.01; Compare with model group, ^▲P＜0.05, ^{▲ ▲}P＜0.01.

The preceding serum BGP content indifference of operation between each group (p〉0.05), administration is after 1 month after the modeling, and model group is apparently higher than normal group (p＜0.01), and three group rules of VAOF control are starkly lower than model group (p＜0.05 ~ p＜0.01).

The clinical observation of embodiment 5 VAOF treatment osteoporosis

1. clinical data

(1) Western medicine diagnose standard: the diagnosing osteoporosis standard adopts and compares with local women's bone density peak value, and reducing 2 standard deviations (SD) above is osteoporosis.This research is with reference to China adult lumbar spine bmd value, and women's peak value is 1.004 ± 0.107g/cm ², at 30 ~ 30 years old age bracket, standard deviation was 0.107, so with 0.790 g/cm ²Be the diagnosing osteoporosis standard, be lower than this value and be sufferers of osteoporosis face.Except secondary osteoporosis, idiopathic osteoporosis disease and took in 3 months and influence bone metabolism medicine person.

(2) tcm diagnosis standard: 1.Ache of the spinal column; 2.The shin knees soreness, hyposexuality; 3.Hiccough and deaf; 4.Shallow complexion; 5.The loss of hairs luxated tooth; 6.Spontaneous sweating; 7.The nocturia frequency; 8.Thready pulse without strength, light red tongue or purplish tongue.Possess above-mentioned 1,2,3 and add 2 persons, examine and be to suffer from a deficiency of the kidney the insufficiency of vital energy and blood card.

(3) case is selected: between in January, 1999 ~ calendar year 2001 December, outpatient service is diagnosed as primary osteoporosis insufficiency of vital energy and blood card patient 32 examples of suffering from a deficiency of the kidney, 50 ~ 80 years old age, average 64.5 years old; 1 ~ 30 year women's menopause time, average 12.8.Be divided into treatment at random and organize 20 examples, matched group 12 examples, two groups of ordinary circumstances are no significant difference (P〉0.05) relatively, sees Table 22.

Table 22 liang group ordinary circumstance comparison (

± s)

Project	VAOF treatment group	The XIANLING GUBAO JIAONANG matched group
			Age (year)	62.3±12.5	63.8±10.9
The menopause time limit (y)	14.2±9.6	13.5±10.3
			Height (cm)	158.1±9.8	157.9±9.4
Body weight (kg)	54.6±8.6	54.9±8.9

2. method

The treatment group gives VAOF capsule (Fructus Ligustri Lucidi 18, Radix Cirsii Japonici 10, the Rhizoma Atractylodis Macrocephalae 10, Radix Salviae Miltiorrhizae 10, Rhizoma Coptidis 5, Radix Notoginseng 6, the Cortex Eucommiae 10 and Fructus Citri Sarcodactylis 10 are provided by the Chinese medicine academy of Guangdong Pharmaceutical University), lot number: 100505), each 3, every day 2 times, serve on 6 months.Matched group: (Guizhou Tongjitang Pharmaceutical Co., Ltd produces, lot number: 100202) each 3, every day 2 times, serve on 6 months to give XIANLING GUBAO JIAONANG.Two groups of viewing durations are withdrawn other Chinese medicine, the relevant Western medicine with primary disease of analgesic.

Observation index:

(1) BMD becomes the coefficient of variation (CV)＜0.1% with Korea S Osteosys borne densitometers DEXXUM3() respectively at reaching treatment detects lumbar vertebra normotopia (L2-L4) after 6 months bone density before the treatment.

(2) Bone Gla protein (BGP) is measured with the RIA method.

(3) urine calcium and urine creatine are measured: adopt fasting urine mensuration in early morning, and calculate Ca/Cr ratio, test kit is provided by middle living company, and ADVIL 1650 automatic clinical chemistry analyzers are measured.

(4) estradiol (E ₂) measure with the RIA method.

(5) clinical manifestation adopt scoring method (do not have card shape note 0 minute, the card shape light or the time and occur remembering 1 fen, symptom occur repeatedly or the time heavily remember 2 fens when light, the card shape continues to occur or severe patient note 3 minutes, the summation of keeping the score that changes before and after the record Syndrome in TCM).

(6) observe untoward reaction.

Curative effect determinate standard

(1) produce effects: the more preceding raising of BMD value is more than 5%, and biochemical indicator obviously improves, and the Syndrome in TCM total mark descends 〉=2/3.

(2) effective: the more preceding raising 2% ~ 4% of BMD value, biochemical indicator makes moderate progress, and the Syndrome in TCM total mark descends 〉=1/3.

(3) invalid: the more preceding raising of BMD value＜2% or continuation descend, and biochemical indicator does not improve or improve not obvious, and the Syndrome in TCM total mark descends＜1/3.

Statistical procedures: adopt X2 check or t check

3. result

(1) clinical efficacy evaluation, the result is shown in table 23, and treatment group and matched group relatively do not have significant difference (P〉0.05).

Table 23 clinical efficacy relatively

Group	The example number	Produce effects (example)	Effectively (example)	Invalid (example)	Total effective rate (%)
						The treatment group	20	7	10	3	85.0%
Matched group	18	7	8	3	83.3%

(2) bone density change to be observed, and the result is shown in table 24: the treatment group all increases before treating with the BMD value treatment back of matched group, more also there was no significant difference between two groups.

The variation of BMD before and after the table 24 liang group treatment (

± s)

Group	The example number	Before the treatment	After the treatment
				The treatment group	20	0.658±0.103	0.761±0.098
Matched group	12	0.663±0.105	0.759±0.099

(3) biochemical written instructions variation is observed, and the result is as shown in Table 25, more slightly reduces by the two groups of E in treatment back before the two groups of BGP in treatment back treat ₂Level is preceding than all be significantly increased (p＜0.05) with treatment, and the ratio of Ca/Cr has tangible reduction.

Table 25 changes of biochemical indexes comparison sheet (

± s)

Group	Before the treatment of treatment group	After the treatment of treatment group	Before the treatment of control group	After the treatment of control group
					E 2(pg.ml-1)	50.61±19.01	66.41±18.12 *	51.23±20.11	65.12±16.45
BGP(ng.ml-1)	6.75±1.81	6.95±1.19	6.88±1.78	6.66±1.83
					Ca/Cr	0.188±0.061	0.098±0.039 *	0.185±0.103	0.134±0.041 *

Annotate: compare before treating the back and treating: ^*P＜0.5.

4. conclusion

Above testing result shows, the VAOF E in the serum that can raise ₂Level reduces Ca/Cr, and treatment back BMD also increases, and can show the alleviation clinical symptoms, and total effective rate reaches 88.5%; Do not have good reaction is obviously arranged, suitable with XIANLING GUBAO JIAONANG, but the curative effect of alleviation back, extremity osteodynia is obvious than XIANLING GUBAO, shows that VAOF can treat osteoporosis effectively, improves symptoms such as the empty blood stasis of bone.

More than to provided by the present invention by Chinese patent 200410051250.4 disclosed a kind of medicines for the treatment of hyperlipemia, by Fructus Ligustri Lucidi, the Rhizoma Atractylodis Macrocephalae, the Cortex Eucommiae, Radix Notoginseng, Radix Cirsii Japonici, Radix Salviae Miltiorrhizae, Rhizoma Coptidis, Fructus Citri Sarcodactylis eight flavor Chinese drug-treated group become compositions, be described in detail in application of medicine that prevents and/or treats the osteoporosis relevant disease and/or health product and preparation method thereof, having used specific case herein sets forth principle of the present invention and embodiment, the explanation of above embodiment just is used for helping to understand method of the present invention and core concept thereof, simultaneously, for one of ordinary skill in the art, according to thought of the present invention, part in specific embodiments and applications all can change, in sum, this description should not be construed as limitation of the present invention.

Claims (3)

1. the application of herbal mixture in preventing and treating the osteoporosis disease, described herbal mixture is to be made by the raw material of Chinese medicine of following percentage by weight: Fructus Ligustri Lucidi 15 ~ 35%, Radix Cirsii Japonici 10 ~ 25%, the Rhizoma Atractylodis Macrocephalae 5 ~ 20%, Radix Salviae Miltiorrhizae 5 ~ 20%, Rhizoma Coptidis 5 ~ 15%, Radix Notoginseng 5 ~ 15%, the Cortex Eucommiae 5 ~ 15% and Fructus Citri Sarcodactylis 5 ~ 15%.

2. the application of herbal mixture according to claim 1 in preventing and treating the osteoporosis disease is characterized in that described osteoporosis disease is the osteoporosis that the osteoporosis that causes of primary osteoporosis, secondary osteoporosis, hyperlipemia or carbohydrate metabolism syndrome cause.

3. the application of herbal mixture according to claim 1 in preventing and treating the osteoporosis disease is characterized in that the osteoporosis that sclerotin is dredged or glucocorticoid causes that described osteoporosis disease is the osteoporosis that causes of hyperlipemia, the carbohydrate metabolism syndrome causes.

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