Goal of the invention
The purpose of this invention is to provide a kind of littler Chinese medicine composition of endometriosis better efficacy side effect for the treatment of.
Another object of the present invention provides the preparation method of this Chinese medicine composition.
Summary of the invention
Medicine of the present invention is made up of effective ingredient and/or pharmaceutically acceptable excipient, and wherein said effective ingredient is to be made by the raw material of Chinese medicine of following consumption:
Rhizoma Curcumae 8-15%, rhizoma sparganic 8-15%, Radix Salviae Miltiorrhizae 14-20%, Radix Curcumae 10-15%, Carapax Trionycis 14-20%, Radix Paeoniae Rubra 14-20%, Hirudo 2-5% and Fructus Aurantii 10-15%.
In order to obtain better therapeutic, the consumption of preferred raw material of Chinese medicine is: Rhizoma Curcumae 9-12%, rhizoma sparganic 9-12%, Radix Salviae Miltiorrhizae 15-18%, Radix Curcumae 11-14%, Carapax Trionycis 15-18%, Radix Paeoniae Rubra 15-18%, Hirudo 3-4% and Fructus Aurantii 11-14%.
In order to obtain optimum curative effect, more preferably the consumption of raw material of Chinese medicine is: Rhizoma Curcumae 10%, rhizoma sparganic 10%, Radix Salviae Miltiorrhizae 16.7%, Radix Curcumae 13.3%, Carapax Trionycis 16.7%, Radix Paeoniae Rubra 16.7%, Hirudo 3.3% and Fructus Aurantii 13.3%.
The method for preparing medicine of the present invention can be Rhizoma Curcumae, rhizoma sparganic, Radix Salviae Miltiorrhizae, Radix Curcumae, Carapax Trionycis, Radix Paeoniae Rubra, Hirudo and the Fructus Aurantii that takes by weighing above-mentioned consumption, and directly with the dry levigation of raw material of Chinese medicine difference, mixing is made powder.Also above-mentioned raw material of Chinese medicine can be extracted and refining carry or the 60-80% ethanol extraction, reclaim or concentrated, make various dosage forms such as capsule, tablet, granule, oral liquid etc. with pharmaceutically acceptable excipient as water.
Chinese medicine composition of the present invention is further to improve prescription on the basis of cowherb rib mixture, rhizoma sparganic, Rhizoma Curcumae, Radix Salviae Miltiorrhizae, Radix Paeoniae Rubra, Bulbus Fritillariae Thunbergii, Carapax Trionycis, Radix Curcumae, Fructus Aurantii, Endothelium Corneum Gigeriae Galli, Radix Angelicae Sinensis and Hirudo 11 flavors are reduced Bulbus Fritillariae Thunbergii, Endothelium Corneum Gigeriae Galli and Radix Angelicae Sinensis, become rhizoma sparganic, Rhizoma Curcumae, Radix Salviae Miltiorrhizae, Radix Curcumae, Radix Paeoniae Rubra, Carapax Trionycis, Fructus Aurantii and Hirudo 8 flavors, wherein monarch drug is Rhizoma Curcumae and rhizoma sparganic; Ministerial drug is Radix Salviae Miltiorrhizae and Radix Curcumae; Adjuvant drug is Carapax Trionycis, Radix Paeoniae Rubra and Hirudo; Messenger drug is a Fructus Aurantii.Increase the consumption of monarch drug, strengthened blood circulation promoting and blood stasis dispelling, reduced the untoward reaction that brings thus with the function of Xiao Disorder eliminating stagnation reduces the consumption of adjuvant drug Hirudo.Through the adjustment of prescription, medicine flavour of a drug of the present invention are simplified, and effect is strengthened, and the effective percentage of process clinical verification treatment endometriosis rises to 90.8% by the effective percentage 80% of cowherb rib mixture, and untoward reaction obviously reduces.
Medicine of the present invention has the effect of blood circulation promoting and blood stasis dispelling, circulation of qi promoting, Xiao Disorder eliminating stagnation, it is pain relieving by vital energy regualting and blood circulation-promoting, blood stasis-eliminating and stagnation-dissipating Er Xiao Disorder, resolving depression regulating menstruation and gravidity assisting, can significantly reduce endometriosis patient's blood viscosity, plasma fibrinogen, packed cell volume, thereby improve pelvic cavity circulation.Compare with the Western medicine danazol, improve dysmenorrhea effect and endometriosis patients serum EMAb negative conversion rate there was no significant difference, but make the pregnancy rate of infertile patient be relatively higher than danazol, and this medicine takes side effect such as no liver function damage for a long time, thereby be better than danazol.
Specific embodiment
Embodiment 1: capsular preparation
Take by weighing Rhizoma Curcumae 9g, rhizoma sparganic 9g, Radix Salviae Miltiorrhizae 15g, Radix Curcumae 12g, Carapax Trionycis 15g, Radix Paeoniae Rubra 15g, Hirudo 3g and Fructus Aurantii 12g, be dried respectively, levigation, mix homogeneously incapsulates.
Embodiment 2: the preparation of tablet
Take by weighing Carapax Trionycis 16g and decoct with water 20 minutes, add Rhizoma Curcumae 10g, rhizoma sparganic 8g, Radix Salviae Miltiorrhizae 14g, Radix Curcumae 15g, Radix Paeoniae Rubra 17g, Hirudo 3g and Fructus Aurantii 17g again, decoct jointly twice, collecting decoction is condensed into thick paste, adds starch and granulates, add magnesium stearate, tabletting, coating.
Embodiment 3: the preparation of granule
Take by weighing Rhizoma Curcumae 8g, rhizoma sparganic 15g, Radix Salviae Miltiorrhizae 16g, Radix Curcumae 13g, Carapax Trionycis 15g, Radix Paeoniae Rubra 15g, Hirudo 3g and Fructus Aurantii 15g, add 70% alcohol reflux 10-20 hour, reclaim ethanol, be condensed into thick paste, add starch and granulate.
The effect of 1 pair of rat experiment temper of test example endometriosis disease
1 material:
Animal: the female sexual maturity of cleaning level is the Wistar rat of copulation not, and body weight 180-220g is provided by Zhongshan Medical Univ.'s Experimental Animal Center.The quality certification number: the moving word 26-98A011 of doctor.Cleaning level environment is raised.
Reagent: TXB
2, 6-keto-PGF
1The α radioimmunological kit is provided by Yangzhou Medical Science College.The IL-2 radioimmunological kit is provided by East Asia, Beijing immunological technique institute.Alsever solution, interim preparation.
Irritate the stomach medication: with medicine of the present invention be mixed with 30% respectively, 7.5% two kind of concentration.Danazol is produced by Yangzhou, Jiangsu pharmaceutical factory, lot number: 970102.Medicine is made into 1% concentration.All preserve standby in 4 ℃ of refrigerators.
Test method
2.1 the foundation of animal model: concrete grammar is as follows: (1) selects the not female rats of copulation of healthy mature for use, regularly carry out vaginal smear examination every day, selected oestrous cycle 4-5 days and had continuously the rat of two normal oestrous cycles, in the rutting period of the 3rd oestrous cycle, with the rat modeling that undergos surgery respectively.(2) operation is under aseptic condition, with 3% pentobarbital sodium 1.5mm/kg intraperitoneal anesthesia, cutting open the belly behind the routine disinfection cuts a cross-talk palace at place, angle, palace, right side, is about 1 centimetre, be placed in the normal saline rapidly and separate, isolated endometrium is cut into two of 5mm * 5mm size, epithelial layer is oppositely facing to body wall, solid on stomach wall with piece of tissue four angle seams, about each one, give gentamycin normal saline solution flushing abdominal cavity before closing abdomen, successively close abdomen at last, cleaning level environment is raised.Cut open the belly once more after (3) 4 weeks and observe the graft upgrowth situation, being masked as successfully of perform the operation: the increase of graft volume, fluid accumulation appears, and mortifier is by connective tissue and omentum majus covering, and vascularization is arranged.Getting graft simultaneously send the real inner membrance of disease probatio inspectionem pecuoarem to survive.
2.2 grouping is a medication: the model winner is divided into four groups at random: model control group, medicine high concentration group of the present invention, medicine low concentration group of the present invention, danazol group.The medication group is the beginning of 31 days after modeling administration all, and wherein the high and low dose group gives 30%, 7.5% concentration liquid respectively, and every day is all by 5.6ml/kg, the danazol group is pressed 4.2ml/kg every day and is irritated stomach, model control group normal saline every day 3ml irritates stomach, and other establishes 10 of normal rats, the conventional raising.Medicine low dose group of the present invention and danazol dosage be all by unit bodies surface area Equivalent Conversion, altogether administration January.
2.3 index detection method: TXB
2, 6-keto-PGF
1α.Measure: get whole blood 4ml and place and contain 2%EDTA-Na
2The anticoagulant plastic test tube in, 4 ℃ of centrifugal separation plasmas are got serum and are placed below-4 ℃ and preserve.Assay method is with reference to the medicine box description.IL-2 measures: get peripheral blood 2ml, after waiting in the injecting tube to solidify, provoke blood coagulation, place 4 ℃ centrifugal, get supernatant and place cryogenic refrigerator to preserve.Assay method is referring to the test kit description.
The peritoneal fluid macrophage phagocytic function is measured: adopt and partly engulf phagocytic index and the phagocytic percentage that method is observed peritoneal macrophage in the chicken red blood cell body.With reference to " guidance of herbal pharmacology experimental technique ", count 200 macrophages, and calculate phagocytic index and phagocytic percentage, the CRBC/ macrophage number (gulping down) that phagocytic index=quilt is engulfed by following formula; Phagocytic percentage=engulf macrophage number/200 macrophage numbers (gulp down and do not gulp down) 100% of CRBC.
Pathological section: routine pathology section preparation method.Cut dystopy endometrium piece, with 10% formaldehyde fixed, conventional dehydration, paraffin embedding, HE dyeing, light microscopy checking.The piece of tissue of getting is all cut into slices in the maximum gauge place.
Electron microscopic observation: the electron microscope specimen preparation method prepares specimen routinely.Piece of tissue is accomplished 1mm
3Fritter is fixed 3 hours in 2.5% glutaraldehyde and 2% paraformaldehyde mixed liquor, after 0.1M phosphoric acid buffer rinsing 3 times, place 1% Osmic acid. solution to fix 1.5 hours; The buffer rinsing of the same method, with the acetone soln ladder dehydration of variable concentrations, Epon812 soaks into, embedding, polymerization; Embedded block is located the back ultrathin section under light microscopic, cut into slices with uranium, plumbous double staining, places the ultrastructure of tissues observed piece under the JEM-1200EX transmission electron microscope.
2.4 statistical method: paired data adopts the t check, rate relatively use u-test.All statistical works all as calculated machine adopt SPSS for Windows (ver 6.0) software to carry out statistical analysis to handle.
3 results
3.1 TXB to endometriosis rat peripheral blood
2, 6-keto-PGF
1α.Influence: the TXB of rat peripheral blood after the modeling
2Content obviously descends (P<0.05), 6-keto-PGF
1α.Content descends more obvious (P<0.01), 6-keto-PGF
1α./ TXB
2Ratio with make film before compare obvious reduction, both compare difference utmost point significance (P<0.01).After the random packet administration, each administration group is compared TXB with model group
2Content all increases, and serves as obvious (P<0.01) with medicine low dose group of the present invention especially wherein; Each administration group 6-keto-PGF
1α.Content all obviously increases (P<0.01), 6-keto-PGF than model group
1α/TXB
2Ratio rise, each administration group is compared difference with model group have significance (P<0.01).The results are shown in Table 1.
Table 1 is respectively organized rat TXB
2, 6-keto-PGF
1The variation comparison of α (X ± s)
Group |
Mus number (only) |
TXB
2(pg/ml)
|
6-keto-PGF
1α (pg/ml)
|
6-keto-PGF
1α /TXB
2 |
Before the modeling |
70 |
572.38±189.68 |
141.87±40.09 |
0.268±0.123 |
After the modeling |
60 |
455.29±158.62 |
54.58±30.50 |
0.143±0.065 |
The high dose of medicine of the present invention |
15 |
543.57±120.52
* |
143.67±20.47
* * |
0.278±0.083
** |
The low dosage of medicine of the present invention |
13 |
572.13±110.13
** |
139.82±54.09
* * |
0.275±0.077
** |
Danazol |
15 |
509.43±104.43
* |
127.67±54.13
* * |
0.262±0.134
** |
Model group |
15 |
427.21±160.96 |
50.59±22.02 |
0.124±0.043 |
With comparison before the modeling, P<0.01, compare with model group,
*P<0.05,
*P<0.01
3.2 influence: peripheral blood IL-2 content obviously raise before than modeling (P<0.01) after the modeling to endometriosis rat peripheral blood IL-2.After the administration, medicine high and low dose group of the present invention and danazol group peripheral blood content obviously descend than model group, analyzing and processing by statistics, and difference all has highly significant (P<0.05), does not have significance (P>0.05) and compare difference before each administration group and the modeling.The results are shown in table 2.
The variation that table 2 is respectively organized rat peripheral blood IL-2 is (X ± s) relatively
Group |
Mus number (only) |
IL-2(ng/ml) |
Before the modeling |
70 |
5.54±1.15 |
After the modeling |
60 |
7.82±1.55 |
The high dose of medicine of the present invention |
15 |
4.74±0.92
* |
The low dosage of medicine of the present invention |
13 |
4.31±0.53
* |
Danazol |
15 |
4.20±0.64
* |
Model group |
15 |
7.79±1.25 |
With comparison before the modeling, P<0.01, compare with model group,
*P<0.05
3.3 the influence to endometriosis rat abdominal cavity macrophage phagocytic function: the peritoneal macrophage phagocytic percentage and the phagocytic index of model control group all obviously increase than normal group, and both compare difference and have utmost point significance (P<0.01).Especially obviously reduce with the peritoneal macrophage phagocytic percentage and the phagocytic index of danazol and high dose group in each group after the administration, comparing difference with model control group has significance (P<0.05).Though the low dose group phagocytic percentage descends to some extent, no statistical significance (P>0.05), its phagocytic index then relatively have obvious reduction (P<0.05) with model control group.The results are shown in Table 3.
The influence of table 3 pair peritoneal macrophage phagocytic function (X ± s)
Group |
Mus number (only) |
Phagocytic percentage (%) |
Phagocytic index |
The normal control group |
10 |
28.1±3.72 |
1.09±0.25 |
Model control group |
10 |
38.4±5.77 |
1.92±0.13 |
The high dose of medicine of the present invention |
15 |
30.3±3.58
* |
1.27±0.44
* |
The low dosage of medicine of the present invention |
13 |
35.2±4.41 |
1.35±0.18 |
Danazol |
15 |
32.4±4.23
* |
1.28±0.33
* |
With comparison before the modeling, P<0.01, compare with model group,
*P<0.05
3.4 influence: the inner membrance survival that visible model group is transplanted under the light microscopic and along the stomach wall superficial growth to endometriosis rat pathological tissue form, form zigzag or annular enclosed structure, the obvious hypertrophy of superficial epithelium becomes column, and have that folliculus is swollen to be formed, body of gland is slot-shaped and high column, and it is cubic that glandular epithelium is, a matter densification, can see a large amount of hemosiderin cells and foam cell in the lumen of gland, rich blood vessel.See after the administration that respectively organizing the Ectopic Endometrium tissue is slot-shaped change, superficial epithelium is short cubic, and cyst is most of to disappear, visible atrophy body of gland, and vascular surface reduces, and Interstitial cell is little and sparse.
3.5 to the Ultrastructural influence of endometriosis rat Ectopic Endometrium: the visible Ectopic Endometrium surface epithelial cell of Electronic Speculum drag group has flourishing microvillus and elongated cilium, abundant organelle is arranged in the endochylema, as Golgi body, ribosome, mitochondrion etc., visible lysosome and secretory granule.And the thin surperficial microvillus of the visible Ectopic Endometrium epithelium of each administration group is short and small sparse, almost can not see cilium, and karyon is big, profile is irregular, and endochylema is few, and the Golgi body in the endochylema, DNA mitochondrial DNA quantity obviously reduce, dilatation of rough endoplasmic reticula, and the granule of taking off phenomenon is arranged, in addition visible a large amount of apoptotic body.Especially change obviously with high dose group and danazol group in each administration group.
Test example 2
Clinical data: endometriosis diagnostic criteria and dysmenorrhea integration standard are all determined by " the new Chinese medicine treatment endometriosis of pelvis clinical research guideline " of Ministry of Public Health promulgation.
Include standard in: allly meet the endometriosis diagnostic criteria and all can include in.
Exclusion standard: (1) is associated with serious disease patients such as hysteromyoma, cardiovascular and cerebrovascular vessel, liver, kidney and hemopoietic system, psychotic; (2) dysmenorrhea that causes by other reasons, infertile or pelvic lump, tuberosity person; (3) do not meet the standard of including in, not medication in accordance with regulations, incomplete, can't judge curative effect person drug allergy or data.
Select endometriosis patient that January to 1998 nineteen ninety-five, year December was confirmed at Guangdong Provincial TCM Hospital endometriosis training totally 97 examples, be divided into treatment group 65 example and matched group 32 examples at random by the prescription on individual diagnosis order.33.4 ± 5.8 years old treatment group mean age; Dysmenorrhea person's 32 examples.Infertile person's 19 examples, pelvic lump, tuberosity person 33 examples.33.7 ± 6.4 years old matched group mean age, dysmenorrhea person's 19 examples.Infertile person's 13 examples, pelvic lump, tuberosity person 24 examples.Credit is analysed by statistics, and two groups of patient's age, symptom distribution situation are all suitable, have comparability (P>0.05) between two groups.
Treatment and observational technique
The treatment group: medicine of the present invention, oral, every day 3 times, each 6; Matched group: danazol (Jiangsu Rieter pharmaceutical factory product) is oral, every day 2 times, each 0.2 gram.Treatment is 6 months continuously.During the treatment, two groups of other medicines of all stopping using.
Observation index: before and after the treatment, two groups of patients' main clinical manifestation is as dysmenorrhea integration, pelvic lump or tuberosity size and infertile patient pregnancy rate; The negative conversion rate of two groups of patients serum EMAb; Treatment group whole blood viscosity, Fibrinogen and hemorheology index change.
Criterion of therapeutical effect and result
" revised standard " by the 3rd academic conference of CAIM (Chinese Association Of Integrative Medicine) department of obstetrics and gynecology Professional Committee.
Result: treatment group 9 examples of fully recovering, produce effects 17 examples, effective 33 examples, invalid 6 examples, cure-remarkable-effectiveness rate 40.0%, total effective rate 90.8%.Matched group 2 examples of fully recovering, produce effects 11 examples, effective 13 examples, invalid 6 examples, cure-remarkable-effectiveness rate 40.6%, total effective rate 81.3%.Two groups of cure-remarkable-effectiveness rates and total effective rate difference do not have significance (P>0.05).
Influence to the endometriosis dysmenorrhea integration: treatment group 30 example is calculated dysmenorrhea integrations before and after the treatment, and is average 10.17 ± 2.29 before the treatment, treatment back 5.34 ± 3.53, and the dysmenorrhea integration difference before and after the treatment of treatment group has utmost point significance (P<0.01).
Influence to endometriosis infertile patient gestation situation: infertile person's 19 examples, 9 examples of becoming pregnant after the treatment are organized in treatment; Infertile person's 13 examples of matched group, 1 example of becoming pregnant after the treatment.Treatment group pregnancy rate is significantly higher than matched group (P<0.05).
Influence to endometriosis patient pelvic lump size (three radial line sums): person's 33 examples that have the enclosed mass in the treatment group, average out to 14.83 ± 4.12cm before treating treats back average out to 11.86 ± 4.38cm; Person's 24 examples that have the enclosed mass in the matched group, average out to 15.76 ± 4.85cm before treating treats back average out to 9.22 ± 4.63cm.The pelvic lump size all has significant difference (P all<0.01) before and after two groups of treatments, but treatment back matched group is significantly less than treatment group (P<0.05).
Influence to endometriosis patients serum EMAb negative conversion rate: positive 27 examples of EMAb before the treatment group is treated, 14 examples of turning out cloudy after the treatment, negative conversion rate 51.9%; Positive 16 examples of EMAb before matched group is treated, 9 examples of turning out cloudy after the treatment, negative conversion rate 56.3%.Two groups of negative conversion rate comparing differences do not have significance (P>0.05).
To the rheol influence of endometriosis blood samples of patients: medicine of the present invention can obviously reduce vital energy stagnation and blood stasis type endometriosis patient whole blood specific viscosity (height is cut) and blood plasma specific viscosity (P all<0.01), reduces fibrinogen content and packed cell volume (P all<0.01) in the blood plasma.See attached list 1.
The variation of hemorheology index before and after the treatment of table 1 treatment group (X ± S)
|
The example number |
Before the treatment |
After the treatment |
Whole blood specific viscosity (height is cut) (mPa.s) |
65 |
5.40±0.59 |
4.56±0.90
* |
Whole blood specific viscosity (low cutting) (mPa.s) |
65 |
7.69±1.23 |
7.22±1.26 |
Blood plasma specific viscosity (height is cut) (mPa.s) |
65 |
1.89±0.21 |
1.77±0.14
* |
Fibrinogen (g/L) |
65 |
3.01±1.25 |
2.51±0.76
* |
Erythrocyte is built up index |
65 |
1.51±0.22 |
1.42±0.25 |
Packed cell volume (%) |
65 |
41.47±3.18 |
38.37±3.97
* |
*With preceding relatively P<0.01 of treatment.