Traditional Chinese medicine composition for improving bone health and preparation and uses thereof

Abstract

A traditional Chinese medicine composition includes 60%-80% by weight of Eucommia ulmoides extract, 15%-30% by weight of Rhizoma Drynariae extract and 5%-10% by weight of Semen Cuscutae extract. A method of preparing the traditional Chinese medicine composition is also provided. Uses of the traditional Chinese medicine composition in the prevention and/or treatment of osteoporosis are also provided.

Claims

1. A method of preparing a traditional Chinese medicine composition for improving bone health, comprising: subjecting Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae to extraction, concentration and drying to produce Eucommia ulmoides extract, Rhizoma Drynariae extract and Semen Cuscutae extract, respectively; and mixing the Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract uniformly to produce the traditional Chinese medicine composition; wherein the method specifically comprises: (1) subjecting Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae to extraction respectively with an ethanol solution under refluxing and filtration; subjecting a residue to extraction with a solvent and filtration; and combining filtrates to obtain a crude Eucommia ulmoides extract, a crude Rhizoma Drynariae extract and a crude Semen Cuscutae extract, respectively; (2) subjecting the crude Eucommia ulmoides extract, the crude Rhizoma Drynariae extract and the crude Semen Cuscutae extract to concentration; introducing maltodextrin to the concentrated Eucommia ulmoides extract and the concentrated Rhizoma Drynariae extract, respectively; wherein the maltodextrin introduced to the concentrated Eucommia ulmoides extract is 5% by weight of Eucommia ulmoides extract; the maltodextrin introduced to the concentrated Rhizoma Drynariae extract is 10% by weight of Rhizoma Drynariae extract; spray drying or vacuum drying and sieving to produce the Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract; and (3) mixing the Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract uniformly in a weight ratio of (60-80):(15-30):(5-10) to produce the traditional Chinese medicine composition.

2. The method of claim 1, wherein in step (1), a volume ratio of Eucommia ulmoides to the ethanol solution is 1:(5-20); a volume ratio of Rhizoma Drynariae to the ethanol solution is 1:(5-20); a volume ratio of Semen Cuscutae to the ethanol solution is 1:(5-20); and the extraction of Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae is performed at 80-100° C. 1-3 times each for 1-3 h, respectively.

3. The method of claim 1, wherein in step (1), a volume ratio of the solvent to the residue is (6-8):1; and the extraction of the residue is performed at 80-100° C. under refluxing 1-3 times each for 1-2 h.

4. The method of claim 2, wherein in step (1), a volume ratio of the solvent to the residue is (6-8):1; and the extraction of the residue is performed at 80-100° C. under refluxing 1-3 times each for 1-2 h.

5. The method of claim 1, wherein in step (2), the spray drying is performed at 180-200° C.; or the vacuum drying is performed at 50-100° C., and the sieving is performed using a sieve of 60-80 mesh.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) The drawings used in the embodiments will be briefly described to make the technical solutions of the disclosure clearer. Obviously, the embodiments illustrated in the drawings are merely part of the embodiments of the disclosure, and other drawings obtained by those skilled in the art without sparing any creative effort should fall within the scope of the invention.

(2) FIG. 1 shows levels of estrogen in rats from different groups.

(3) FIG. 2 illustrates detection results of blood biochemical indexes of rats from different groups.

(4) FIG. 3 illustrates detection results of bone metabolic biochemical markers of rats from different groups.

(5) FIG. 4 illustrates detection results of bone density-associated markers of rats from different groups.

(6) FIG. 5 shows testing results of mechanical indexes of rats from different groups.

DETAILED DESCRIPTION OF EMBODIMENTS

(7) Technical solutions of the present disclosure will be clearly and completely described below with reference to the embodiments. Obviously, described below are merely some embodiments of the disclosure, which are not intended to limit the disclosure. Other embodiments made by those skilled in the art without sparing any creative effort should fall within the scope of the disclosure.

(8) The disclosure provides a traditional Chinese medicine composition, which has no toxic side effects and high adsorption. The composition can improve bone health, and can be used as a dietary supplement or a health food raw material for the prevention or treatment of osteoporosis, and thus it is suitable for commercial promotion.

(9) The invention will be further described below with reference to the embodiments. It should be understood that these embodiments are merely illustrative of the invention, and are not intended to limit the invention. Any improvement and modification made by those skilled in the art without departing from the spirit of the invention should still fall within the scope of the invention.

(10) The disclosure provides a traditional Chinese medicine composition for improving bone health, including:

(11) 60%-80% by weight of Eucommia ulmoides extract;

(12) 15%-30% by weight of Rhizoma Drynariae extract; and

(13) 5%-10% by weight of Semen Cuscutae extract.

(14) In an embodiment, the above extracts are separately extracted from the corresponding traditional Chinese medicine material, and the traditional Chinese medicine composition is prepared by extracting the above extracts respectively from Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae and compounding.

(15) The disclosure also, provides a method of preparing the above traditional Chinese medicine composition, including:

(16) subjecting Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae to extraction, purification, concentration and spray drying to produce Eucommia ulmoides extract, Rhizoma Drynariae extract and Semen Cuscutae extract, respectively; and

(17) mixing the Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract uniformly to produce the traditional Chinese medicine composition;

(18) where the method specifically includes:

(19) (1) subjecting Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae to extraction respectively with an ethanol solution under refluxing and filtration; subjecting a residue to extraction with a solvent and filtration several times; and combining filtrates to obtain a crude Eucommia ulmoides extract, a crude Rhizoma Drynariae extract and a crude Semen Cuscutae extract, respectively;

(20) (2) subjecting the crude Eucommia ulmoides extract, the crude Rhizoma Drynariae extract and the crude Semen Cuscutae extract to purification, concentration, spray drying or vacuum drying and sieving to produce the Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract; and

(21) (3) mixing the Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract uniformly in a weight ratio of (60-80):(15-30):(5-10) to produce the traditional Chinese medicine composition.

(22) In an embodiment, in step (1), a volume ratio of Eucommia ulmoides to the ethanol solution is 1:(5-20); a volume ratio of Rhizoma Drynariae to the ethanol solution is 1:(5-20); a volume ratio of Semen Cuscutae to the ethanol solution is 1:(5-20); and the extraction of Rhizoma Drynariae, Rhizoma Drynariae and Semen Cuscutae is performed at 80-100° C. 1-3 times each for 1-3 h, respectively.

(23) In an embodiment, in step (1), a volume ratio of the solvent to the residue is (6-8):1; and the extraction of the residue is performed at 80-100° C. under refluxing 0-3 times each for 1-2 h.

(24) In an embodiment, in step (2), the spray drying is performed at 180-200° C.; the vacuum drying is performed at 50-100° C.; and the sieving is performed using a sieve of 60-80 mesh.

(25) In an embodiment, step (2) further includes:

(26) before the spray drying or the vacuum drying, introducing a dextrin to the concentrated Eucommia ulmoides product, the concentrated Rhizoma Drynariae product and the concentrated Semen Cuscutae product, respectively, followed by mixing uniformly;

(27) where the dextrin is 1%-20% by weight of Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae, respectively.

(28) In a third aspect, the disclosure further provides a method of treating, osteoporosis in a patient in need thereof, including:

(29) administering, an effective amount of the above traditional Chinese medicine composition to the patient;

(30) In an embodiment, the traditional Chinese medicine composition is used as a dietary supplement or a health-care food; and the traditional Chinese medicine composition is in a form of a capsule, a granule or a tablet.

(31) Technical solutions of the disclosure will be further described below with reference to the embodiments.

Example 1

(32) Provided herein was a method of preparing a traditional Chinese medicine, for improving bone health, which was specifically described as follows.

(33) (1) Eucommia ulmoides was subjected to extraction with a 60%-80% ethanol solution in a volume ratio of 1:6 under refluxing for 2 h and filtration to obtain a residue. The residue was subjected to extraction with deionized water twice in a volume ratio of 1:6 under refluxing each for 1.5 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 5% by weight of Eucommia ulmoides. The combined filtrate was continuously concentrated to a density of 1.15-1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Eucommia ulmoides extract.

(34) (2) Rhizoma Drynariae was crushed and subjected to extraction with a 75% ethanol solution three times respectively in a volume ratio of 1:6, 1:5 and 1:5 under refluxing each for 2 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 10% by weight of Rhizoma Drynariae. Then the combined filtrate was continuously concentrated to a density of 1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Rhizoma Drynariae extract.

(35) (3) Semen Cuscutae was subjected to extraction with a 75% ethanol solution three times respectively in a volume ratio of 1:6, 1:5 and 1:5 under refluxing each for 2 h and filtration. Filtrates were combined, concentrated to a density of 1.12-1.15 and subjected to spray drying at 180° C. The dried product was sieved with a sieve of 80 mesh to produce the Semen Cuscutae extract.

(36) (4) The Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract were mixed uniformly in a weight ratio of 60:30:10 to produce the traditional Chinese medicine composition.

Example 2

(37) Provided herein was a method of preparing a traditional Chinese medicine for improving bone health, which was specifically described as follows.

(38) (1) Eucommia ulmoides was subjected to extraction With a 85% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with deionized water twice in a volume ratio of 1:6 under refluxing each for 1 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 5% by weight of Eucommia ulmoides. The combined filtrate was continuously concentrated to a density of 1.15-1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Eucommia ulmoides extract.

(39) (2) Rhizoma Drynariae was subjected to extraction with a 80% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue. The residue was subjected to extraction with the 80% ethanol solution twice in a volume ratio of 1:6 under refluxing each for 2 h. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 10% by weight of Rhizoma Drynariae. Then the filtrate was continuously concentrated to a density of 1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Rhizoma Drynariae extract.

(40) (3) Semen Cuscutae was immersed in a 80% ethanol solution in a volume ratio of 1:8 for 0.5 h and subjected to extraction under refluxing for 2 h and filtration to obtain, a residue. The residue was subjected to extraction with the 80% ethanol solution in a volume ratio of 1:6 under refluxing twice each for 1 h. Filtrates were combined, concentrated to a density of 1.12-1.15 and subjected to spray drying at 180-200° C. The dried product was sieved with a sieve of 80 mesh to produce the Semen Cuscutae extract.

(41) (4) The Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract were mixed uniformly in a weight ratio of 60:30:10 to produce the traditional Chinese medicine composition.

Example 3

(42) Provided herein was a method of preparing a traditional Chinese medicine for improving bone health, which was specifically described as follows.

(43) (1) Eucommia ulmoides was subjected to extraction With a 70% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with deionized water twice in a volume ratio of 1:6 under refluxing each for 1 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 5% by weight of Eucommia ulmoides. The combined filtrate was continuously concentrated to a density of 1.15-1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Eucommia ulmoides extract.

(44) (2) Rhizoma Drynariae was subjected to extraction with a 80% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution twice in a volume ratio of 1:6 under refluxing each for 2 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 10% by weight of Rhizoma Drynariae. Then the combined filtrate was continuously concentrated to a density of 1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Rhizoma Drynariae extract.

(45) (3) Semen Cuscutae was immersed in a 80% ethanol solution in a volume ratio of 1:8 for 0.5 h and subjected to extraction under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution in a volume ratio of 1:6 under refluxing twice each for 1 h. Filtrates were combined, concentrated to a density of 1.12-1.15 and subjected to spray drying at 180-200° C. The dried product was sieved with a sieve of 80 mesh to produce the Semen Cuscutae extract.

(46) (4) The Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract were mixed uniformly in a weight ratio of 75:15:10 to produce the traditional Chinese medicine composition.

Example 4

(47) Provided herein was a method of preparing a traditional Chinese medicine for improving bone health, which was specifically described as follows.

(48) (1) Eucommia ulmoides was subjected to extraction With a 85% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with deionized water twice in a volume ratio of 1:6 under refluxing each for 1 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 5% by weight of Eucommia ulmoides. The combined filtrate was continuously concentrated to a density of 1.15-1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Eucommia ulmoides extract.

(49) (2) Rhizoma Drynariae was subjected to extraction with a 80% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution twice in a volume ratio of 1:6 under refluxing each for 2 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 10% by weight of Rhizoma Drynariae. Then the combined filtrate was continuously concentrated to a density of 1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Rhizoma Drynariae extract.

(50) (3) Semen Cuscutae was immersed in a 80% ethanol solution in a volume ratio of 1:8 for 0.5 h and subjected to extraction under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution in a volume ratio of 1:6 under refluxing twice each for 1 h. Filtrates were combined, concentrated to a density of 1.12-1.15 and subjected to spray drying at 180-200° C. The dried product was sieved with a sieve of 80 mesh to produce the Semen Cuscutae extract.

(51) (4) The Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract were mixed uniformly in a weight ratio of 70:20:10 to produce the traditional Chinese medicine composition.

Example 5

(52) Provided herein was a method of preparing a traditional Chinese medicine for improving bone health, which was specifically described as follows.

(53) (1) Eucommia ulmoides was subjected to extraction With a 85% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with deionized water twice in a volume ratio of 1:6 under refluxing each for 1 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 5% by weight of Eucommia ulmoides. The combined filtrate was continuously concentrated to a density of 1.15-1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Eucommia ulmoides extract.

(54) (2) Rhizoma Drynariae was subjected to extraction with a 80% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution twice in a volume ratio of 1:6 under refluxing each for 2 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 10% by weight of Rhizoma Drynariae. Then the combined filtrate was continuously concentrated to a density of 1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Rhizoma Drynariae extract.

(55) (3) Semen Cuscutae was immersed in a 80% ethanol solution in a volume ratio of 1:8 for 0.5 h and subjected to extraction under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80%) ethanol solution in a volume ratio of 1:6 under refluxing twice each for 1 h. Filtrates were combined, concentrated to a density of 1.12-1.15 and subjected to spray drying at 180-200° C. The dried product was sieved with a sieve of 80 mesh to produce the Semen Cuscutae extract.

(56) (4) The Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract were mixed uniformly in a weight ratio of 80:5:5 to produce the traditional Chinese medicine composition.

Example 6

(57) Provided herein was a method of preparing a traditional Chinese medicine for improving bone health, which was specifically described as follows.

(58) (1) Eucommia ulmoides was subjected to extraction With a 85% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with deionized water twice in a volume ratio of 1:6 under refluxing each for 1 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 5% by weight of Eucommia ulmoides. The combined filtrate was continuously concentrated to a density of 1.15-1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Eucommia ulmoides extract.

(59) (2) Rhizoma Drynariae was subjected to extraction with a 80% ethanol solution in a volume ratio of 1:8 under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution twice in a volume ratio of 1:6 under refluxing each for 2 h and filtration. Filtrates were combined, concentrated and added with maltodextrin, where the maltodextrin was 10% by weight of Rhizoma Drynariae. Then the combined filtrate was continuously concentrated to a density of 1.2, dried at 75° C. under vacuum and sieved with a sieve of 80 mesh to produce the Rhizoma Drynariae extract.

(60) (3) Semen Cuscutae was immersed in a 80% ethanol solution in a volume ratio of 1:8 for 0.5 h and subjected to extraction under refluxing for 2 h and filtration to obtain a residue and a filtrate. The residue was subjected to extraction with the 80% ethanol solution in a volume ratio of 1:6 under refluxing twice each for 1 h. Filtrates were combined, concentrated to a density of 1.12-1.15 and subjected to spray drying at 180-200° C. The dried product was sieved with a sieve of 80 mesh to produce the Semen Cuscutae extract.

(61) (4) The Eucommia ulmoides extract, the Rhizoma Drynariae extract and the Semen Cuscutae extract were mixed uniformly in a weight ratio of 65:30:5 to produce the traditional Chinese medicine composition.

Example 7

(62) Provided herein was a method of detecting the activity of per unit of individual extract in the traditional Chinese medicine composition for improving bone health, which was specifically described as follows.

(63) The total flavonoids in the extracts were determined by ultraviolet spectrophotometry.

(64) Preparation of Reference Sample

(65) 10 mg, of rutin standard was accurately weighed, transferred to a 50 mL volumetric flask, added with methanol to the volume and mixed uniformly to produce a 0.2 mg/mL mother liquor.

(66) 0 mL, 1 mL, 2 mL, 4 mL and 8 mL of the mother liquor were respectively added to five 25 mL test tubes with a stopper and diluted with pure water to 10 mL.

(67) Preparation of Test Sample

(68) 0.1 g of each extract (Eucommia ulmoides, Rhizoma Drynariae and Semen Cuscutae) was accurately weighed, dissolved with 50 mL of methanol and subjected, to ultrasonication for 1 h to produce an extract solution. 3 mL of the extract solution was transferred to a 25 mL test tube with a stopper and diluted to 10 mL with water.

(69) Each reference sample and the test sample were respectively added with 1 mL of 5% NaNO.sub.2 solution and subjected to standing for 6 min. Then each, reaction mixture was added with 1 mL of a 10% Al(NO.sub.3).sub.3 solution, subjected to standing for 6 min, added with 1 mL of a 1 mol/L NaOH solution and diluted to 25 mL with distilled water for determination.

(70) Test results were listed in detail in Tables 1 and 2, where an initial concentration of the rutin reference sample (purity: 94.9%) was 0.2076 mg/mL.

(71) TABLE-US-00001 TABLE 1 Absorbance(Abs) of reference samples Volume of reference Concentration samples (mL) (mg/mL) Abs 1 0.0197 0.093 2 0.0393 0.193 4 0.0787 0.403 8 0.1574 0.843

(72) A standard curve was plotted with y=5.4633×−0.02 and R.sup.2=0.9997.

(73) TABLE-US-00002 TABLE 2 Absorbance and concentration of test samples Concen- Per- tration Average centage Sample Weight/g Abs (mg/g) (mg/g) (%) Eucommia ulmoides 0.1319 0.421 102.00 101.77 10.18 extract 0.419 101.53 Rhizoma Drynariae 0.1361 0.239 58.05 57.72 5.77 extract 0.236 57.38 Semen Cuscutae 0.1375 0.285 67.67 67.67 6.77 extract 0.285 67.67

(74) Described below were the common preparations of the traditional Chinese medicine composition provided herein when applied to the preparation of health food.

Example 8

(75) The Eucommia ulmoides extract, Rhizoma Drynariae extract and Semen Cuscutae extract prepared in Example 1 were mixed uniformly and sieved with a sieve of 80 mesh to produce a traditional Chinese medicine composition. Then the composition was mixed uniformly with starch and magnesium stearate, pressed into tablets and film-coated to produce final tablets.

Example 9

(76) The liquid extracts prepared in Example 1 were mixed uniformly, decoded to 1 g/mL, and transferred to a beaker, to which sucrose, starch and dextrin Were introduced and mixed uniformly to produce a mixture. When the mixture was kneaded into a dough and dispersed when touched, granules that can not pass through the 20-mesh sieve and 80-mesh sieve but can pass the 10-mesh sieve Were collected and dried in an oven in time for use.

(77) Finally, the collected granules Were dried under Vacuum to a water content of 4% to produce the traditional Chinese medicine granules.

Example 10

(78) The extracts prepared in Example 1 were mixed uniformly and sieved with a 80-mesh sieve to produce a traditional Chinese medicine composition, which was further mixed uniformly with magnesium stearate and encapsulated at a filling amount of 0.53-0.54 g to obtain a traditional Chinese medicine capsule.

(79) Embodiments provided herein are described in a progressive manner, and each embodiment mainly focuses on the differences from other embodiments. The same or similar parts among various embodiments can be referred to each other.

(80) Further, the following test was performed to evaluate the efficacy of the traditional Chinese medicine composition prepared herein.

(81) Efficacy Test

(82) Samples: the traditional Chinese medicine (TCM) composition prepared in Example 1, a positive drug (Estradiol Valerate) and a competing drug (Caltrate).

(83) Grouping

(84) Female SD rats were randomly divided into sham operation group, model group, positive control group (estradiol valerate), competing drug group (Caltrate), high dose TCM composition (prepared in Example 1) group and low dose TC′M composition (prepared in Example 1) group, and 8 in each group.

(85) Model Establishment

(86) Female SD rats were treated by bilateral ovariectomy to establish osteoporosis model rats.

(87) Administration

(88) The osteoporosis model rats were subjected to intragastric administration once a day and the administration cycle lasted for 8 weeks.

(89) Indicators

(90) At the end of each week, each rat was fasted (water was allowed) for 12 hours to measure the weight change, observe and record the eating and mental state and the coat color and evaluate the quality of the model. Further, the vaginal epithelial cells were subjected to smear test to obtain the estrus cycle to determine whether the rats were completely ovariectomized.

(91) After 8 weeks, the rats were sacrificed, and blood was collected from the abdominal aorta and subjected to standing for 1 hour. Then the blood sample was centrifuged to collect serum, which was detected for the bone metabolic biochemical indicators, including bone glaprotein (BGP), bone specific alkaline phosphatase (BALP), carboxyterminal propeptide of type I procollagen (PICP) and N-telopeptides of type I collagen (NTX); estrogen indicators, including serum estradiol E2 and follicle stimulating hormone (FSH); and blood biochemical indexes, including serum phosphorus and serum calcium. Moreover, the complete bilateral femurs were collected, on which the muscle tissues were, completely removed. The left femur was used for femoral mechanical testing, and the right femur was tested for related parameters such as bone density. The above experimental data was used to evaluate the efficacy of the TCM composition of the disclosure.

(92) It should be noted that the dosage for each test group was shown as follows.

(93) The dosage of the TCM composition was determined based on the recommended crude drug dosage in the pharmacopoeia, and 1 g of extract was obtained after the extraction.

(94) Positive control group: 0.1 g/day for human.

(95) Competing drug group (Caltrate): one tablet (1 g)/day and 600 mg of calcium/tablet.

(96) High dose TCM composition group: 1 g/day for human; low dose TCM composition group: 0.5 g/day for human.

(97) The dosage for rats was calculated as follows: dosage for rat (daily dose for human/60)*6;

(98) where 60 referred to body weight of human, and 6 indicated the rat coefficient.

(99) The experimental results were presented as follows.

(100) TABLE-US-00003 TABLE 3 Serum estrogen indicators of rats in different groups Dosage E2 FSH Group (mg/kg) (pg/mL) (ng/mL) Sham-operation / 67.69 ± 18.74#  57.71 ± 25.62# Model / 37.96 ± 11.37* 109.22 ± 22.25* Positive control 10 32.45 ± 10.24*  96.55 ± 11.84* Caltrate 60 39.17 ± 6.97*  118.02 ± 35.68* High dose TCM 100 40.16 ± 8.82*  111.09 ± 25.16* in Exa. 1 Low dose TCM 50  50.59 ± 13.76*#  98.78 ± 25.22* in Exa. 1 Notes: *indicated that there was significant difference (P < 0.05) when compared to the sham-operation group; and #indicated that there was significant difference (P < 0.05) when compared to the model group.

(101) The corresponding histogram was shown in FIG. 1. After the 8 weeks administration, the ELISA test results of the two estrogen indicators (serum estradiol E2 and FSH) revealed that compared to the model group, the low dose TCM composition group had a significantly higher E2 level (P<0.05). Therefore, it can be demonstrated that the TCM composition of the disclosure can significantly inhibit the decline of the serum E2 level in rats, and had the effect of preventing the reduction of estrogen level in menopause, preventing the bone loss.

(102) TABLE-US-00004 TABLE 4 Serum biochemical indexes of rats in different groups Dosage P Ca Group (mg/kg) (mmol/L) (mmol/L) Sham-operation / 2.43 ± 0.59 2.42 ± 0.12 Model / 2.38 ± 0.44 2.26 ± 0.25 Positive control 10   3.1 ± 0.53*# 2.33 ± 0.13 Caltrate 60 2.34 ± 0.35  2.4 ± 0.09 High dose TCM 100  1.9 ± 0.39* 2.31 ± 0.1  in Exa. 1 Low dose TCM 50 2.23 ± 0.57 2.19 ± 0.3  in Exa. 1 Notes: *indicated that there was significant difference (P < 0.05) when compared to the sham-operation group; and #indicated that there was significant difference (P < 0.05) when compared to the model group.

(103) The corresponding histogram was shown in FIG. 2. After the 8 weeks' administration, the positive control group had significantly higher serum phosphorus content than the model group and the sham-operation group (P<0.05), while the serum phosphorus content of the high dose TCM composition group was significantly lower than the sham-operation group (P<0.05). Other groups had similar serum phosphorus content. There was no significant difference among all experimental groups with respect to the serum calcium content.

(104) TABLE-US-00005 TABLE 5 Bone metabolic biochemical indicators of rats in different groups Dosage BGP BALP PICP NTX Group (mg/kg) (ng/mL) (pg/mL) (ng/mL) (ng/mL) Sham-operation / 10.53 ± 1.78  861.31 ± 123.6 743.78 ± 94.67  23.31 ± 6.41 Model / 9.97 ± 1.76 791.63 ± 98.35 686.71 ± 95.49  24.27 ± 3.53 Positive control 10 9.93 ± 2.23  764.2 ± 123.81 739.33 ± 175.34 26.92 ± 5.68 Caltrate 60 9.11 ± 1.29 762.45 ± 79.35 797.25 ± 136.34 23.64 ± 2.58 High dose TCM 100 10.57 ± 1.12   770.2 ± 129.17 747.88 ± 173.12 26.17 ± 2.4  in Exa. 1 Low dose TCM 50 11.1 ± 1.24  791.45 ± 106.44 795.06 ± 104.06 25.51 ± 3.96 in Exa. 1 Notes: * indicated that there was significant difference (P < 0.05) when compared to the sham-operation group; and # indicated that there was significant difference (P < 0.05) when compared to the model group.

(105) The corresponding histogram, was shown in FIG. 3. After the 8 weeks' administration, the ELISA test results showed that there was no significant difference among all experimental groups with respect to the four bone metabolic indicators (BGP, BALP, PICP and NTX).

(106) TABLE-US-00006 TABLE 6 Bone density parameters of rats in different groups Dosage BMD BV/TV Tb. N Tb. Sp Group (mg/kg) (g/cm.sup.2) (%) (1/mm) (μm) Sham-operation / 0.257 ± 0.057.sup.#  22.5 ± 4.sup.#   .sup. 3.25 ± 0.571.sup.# 258.837 ± 77.079.sup.#   Model / 0.186 ± 0.049* 15.56 ± 4.14* 2.195 ± 0.53*  500.657 ± 155.844* Positive control 10 0.198 ± 0.051* 16.863 ± 5.237* 2.392 ± 0.809*  455.6 ± 182.901* Caltrate 60 0.165 ± 0.048* 13.826 ± 4.365* 1.983 ± 0.588* 535.106 ± 176.152* High dose TCM 100 0.246 ± 0.065.sup.#  20.083 ± 5.255  2.758 ± 0.711  346.449 ± 116.986  in Exa. 1 Low dose TCM 50 0.181 ± 0.049* 15.582 ± 4.905* 2.248 ± 0.719* 492.541 ± 178.987* in Exa. 1 Notes: *indicated that there was significant difference (P < 0.05) when compared to the sham-operation group; and .sup.#indicated that there was significant difference (P < 0.05) when compared to the model group.

(107) The corresponding histogram was shown in FIG. 4. It can be seen through the analysis of parameters of the bone tissue microstructure at the femoral metaphysis region of rats that compared to the sham-operation group, the model group had significantly lower bone mineral density (BMD), bone volume fraction (BV/TV) and (Tb.N) (P<0.05) and significantly higher trabecular separation/spacing (Tb.Sp) (P<0.05); compared to the model group, the high dose TCM composition group had significantly increased bone mineral density (P<0.05), and exhibited an increasing trend in the bone volume fraction and the trabecular number and a decreasing trend in the trabecular separation/spacing. The above results demonstrated that the composition provided herein can improve the bone density, and thus it had a certain, preventive effect on osteoporosis.

(108) TABLE-US-00007 TABLE 7 Mechanical indexes of rats in different groups Group Dosage (mg/kg) Load/N Sham-operation / 138.16 ± 26.71# Model /  93.13 ± 17.74* Positive control 10 109.67 ± 26.78* Caltrate 60 104.06 ± 19.52* High dose TCM 100  126.6 ± 12.57# in Exa. 1 Low dose TCM 50  89.98 ± 23.41* in Exa. 1 Notes: *indicated that there was significant difference (P < 0.05) when compared to the sham-operation group; and #indicated that there was significant difference (P < 0.05) when compared to the model group.

(109) The corresponding histogram was shown in FIG. 5. It can be seen from the biomechanical test results of rat femurs that compared to the sham-operation group, the model group had significantly lower maximum load (P<0.05); the high dose TCM composition group was significantly higher than the model group in the maximum load (P<0.05). The above results indicated that the composition prepared herein, can reduce the risk of fracture and had a certain preventive effect on the osteoporosis.

(110) Based on the analysis of the above experimental results, it can be concluded that compared with the model group, the composition provided herein can significantly inhibit the decrease in the serum E2 level, increase bone density and the maximum load of femur in rats. Therefore, the composition of the disclosure had the effects of preventing the decline of estrogen level in menopause, preventing the sexual organ atrophy, improving sexual function, inhibiting bone loss and increasing the bone density and can be used to prevent osteoporosis and improve bone health.

(111) Described above are merely illustrative of the invention to enable those skilled in the art to implement or use the invention, and are not intended to limit the invention. It should be understood that any modification, replacement and change made by those skilled in the art without, departing from the spirit of the invention should fall within the scope of the invention.