Summary of the invention
The purpose of this invention is to provide the application of a kind of extractive of general flavone that from Herba Elsholtziae Blandae, extracts in preparation anti-cardiovascular disease medicine.
Medicine of the present invention has better action for anti-cardiovascular disease-myocardial ischemia, arrhythmia, blood fat reducing etc., have class estrogen and pregnancy hormone like-effect simultaneously, be used for climacteric syndrome, osteoporosis, neurodegenerative diseases and have effects such as eliminating phlegm and stopping cough, anti-capillary fragility and unusual permeability, inhibition tumor cell, spasmolytic, follicle hormone-like function, removing free radical.
The present invention extracts extractive of general flavone from the medicinal plants Herba Elsholtziae Blandae, in extractive of general flavone: adjuvant=60~99: 40~1 ratio is made medicament.
Medicine of the present invention is that the Herba Elsholtziae Blandae plant is is further researched and developed, extracted the effective site in the Herba Elsholtziae Blandae---the new purposes of total flavones, found that it has therapeutical effect preferably to anti-cardiovascular disease, as myocardial ischemia, arrhythmia, myocardial infarction.In addition, also have class estrogen and pregnancy hormone like-effect, be used for climacteric syndrome, osteoporosis, neurodegenerative diseases and have effects such as eliminating phlegm and stopping cough, anti-capillary fragility and unusual permeability, inhibition tumor cell, spasmolytic, follicle hormone-like function, removing free radical.
Below curative effect of medication of the present invention is described with pharmacodynamic experiment of the present invention.
Pharmacodynamic experiment method of the present invention and result
One. bringing out the Acute Myocardial Ischemia in Rats test with the anti-pituitrin of the present invention is example
81 of SD male rats, be divided into 5 groups at random, the basic, normal, high dosage group of JGS is igJGS extract 25,50,100 μ g/kg (in general flavone content) respectively, other establishes solvent control group ig4ml/kg0.5%CMC solution, positive controls ig DIAOXINXUE KANG 150mg/kg (in the total Saponin of steroidal).The above-mentioned all continuous gastric infusion of rat 5 days of respectively organizing, irritated in the 5th day behind the stomach 40 minutes, the anesthesia of lumbar injection 2g/kg25% urethane, trace one section II continuously by Medlab5.5 bio signal acquisition system and lead electrocardiogram, 60 minutes tail vein injection pituitrin 0.75U/kg behind the filling stomach, constant speed pushed away in 10 seconds, continuous record electrocardio Figure 10 minute.Preceding and the injection electrocardiogram variation in the time of back 15 seconds, 30 seconds, 1,2,3,5,7,10 minute of mensuration injection respectively, ecg measurement is a baseline with the PR section, and each time point is surveyed 5 cardiac cycles, gets its meansigma methods, the variation of T wave-amplitude and heart rate between comparable group, and the credit that takes statistics is analysed.
Experiment finishes back rat broken end gets blood, cores dirtyly, adopts Olympus Au-2700 automatic clinical chemistry analyzer to measure serum CK, Ca
2+, adopt nitrate reductase method NO test kit (Wenzhou, Zhejiang p020513 of health Bioisystech Co., Ltd of Erie) to measure NO.Heart separates left ventricle and weighs with 4 ℃ of clean remained blood of normal saline flushing, adds 4 ℃ of normal saline of 9 times of weight and makes 10% myocardium homogenate, measures homogenate MDA content with the TBA method, and the Coomassie brilliant blue method is measured protein content, gets the homogenate supernatant and measures Ca
2+And NO, the same serum of method.
Complex electrocardio figure and biochemical indicator are estimated the effect of medicine.
Table 1 JGS brings out the influence (x ± S) of rat ventricular to pit
Group | Dosage (μ g/kg) | Number of animals (only) | Incidence of arrhythmia % |
0.5%CMC JGS JGS JGS DIAOXINXUE KANG | - 25 50 100 150 | 17 16 16 15 16 | 82.4(14) 75(12) 68.8(11) 33.3(5)
△△ 68.8(11)
|
Compare with normal group:
△ △P<0.01, x
2Check
Table 2 JGS brings out the influence (x ± S) of rat heart muscle ischemia ECG T wave to pit
Group | Dosage (μ g/kg) | Number of animals (only) | Before the modeling | T ripple (mv) after the modeling |
T ripple (mv) | 30 seconds | The increase and decrease percentage rate | 1 minute | The increase and decrease percentage rate | 3 minutes | The increase and decrease percentage rate | 7 minutes | The increase and decrease percentage rate |
0.5%CMC JGS JGS JGS Diaoxinxue Kang | - 25 50 100 150 | 17 16 16 15 16 | 0.188± 0.042 0.204± 0.037 0.202± 0.048 0.235± 0.040 0.206± 0.034 | 0.224± 0.090 0.217± 0.082 0.216± 0.078 0.248± 0.069 0.214± 0.108 | 45.1± 32.1 43.2± 30.5 34.8± 27.8 27.3± 20.4
△ 45.7± 22.7
| 0.185± 0.078 0.190± 0.067 0.176± 0.065 0.193± 0.099 0.218± 0.059 | -3.7± 29.5 -5.4± 31.5 -10.9± 30.6 -18.1± 40.6 4.8± 18.4 | 0.161± 0.060 0.188± 0.051 0.158± 0.074 0.158± 0.097 0.202± 0.035 | -14.4± 24.6 -7.1± 21.3 -22.0± 30.1 -30.1± 39.2 -0.2± 18.7 | 0.145± 0.054 0.163± 0.036 0.148± 0.065 0.148± 0.080 0.177± 0.032 | -22.3± 22.6 -18.8± 17.6 -27.0± 28.4 -35.5± 32.2 -12.1± 16.9 |
Compare with normal group:
△P<0.05, rank test
Table 3 JGS brings out the influence (x ± S) of rat heart muscle ischemia heart rate to pit
Group | Dosage (μ g/kg) | Number of animals (only) | Before the modeling | Heart rate after the modeling (inferior/minute) |
Heart rate (inferior/minute) | 30 seconds | The increase and decrease percentage rate | 1 minute | The increase and decrease percentage rate | 3 minutes | The increase and decrease percentage rate | 7 minutes | The increase and decrease percentage rate |
0.5%CMC JGS JGS JGS Diaoxinxue Kang | - 25 50 100 150 | 17 16 16 15 16 | 375± 62.8 406± 83.9 401± 77.8 344± 95.8 386± 86.2 | 283± 48.5 292± 63.7 312± 35.5 274± 57.2 Z70± 53.4 | -23.6± 10.6 -27.3± 10.9 -20.0± 15.4 -16.7± 17.7 -27.4± 16.7 | 277± 58.7 306± 59.4 313± 34.3 284± 46.6 295± 53.5 | -22.2± 12.8 -23.7± 10.4 -20.0± 13.4 -13.7± 16.0 -21.4± 14.7 | 256± 36.4 277± 30.8 278± 30.9 274± 44.3
△ 271± 49.4
| -30.0± 15.2 -29.7± 13.0 -27.9± 18.4 -14.1± 19.6 -26.8± 17.8 | 258± 29.0 270± 39.6 254± 37.7 264± 36.1 260± 48.9 | -29.8± 12.9 -30.9± 16.9 -34.4± -19.2± 16.2
△ -29.6± 16.9
|
Compare with normal group:
△P<0.05, rank test
Table 4 JGS brings out rat heart muscle ischemia serum CK, Ca to pit
2+Influence (x ± S)
Group | Dosage (μ g/kg) | N | CK(U/L) | Ca
2+(mmol/L)
|
0.5%CMC JGS JGS JGS DIAOXINXUE KANG | - 25 50 100 150 | 16 16 15 15 16 | 3299±750.7 2492±1078.5
△ 2491±1093.6
△ 2646±719.8
△ 2849±580.9
△ | 2.28±0.049 2.33±0.093
△ 2.34±0.11
△ 2.37±0.13
△△ 2.34±0.078
△△ |
Compare with normal group:
△P<0.05,
△ △P<0.01, the t check
Table 5 JGS brings out rat heart muscle ischemia serum NO level and myocardium homogenate NO, MDA, Ca to pit
2+Influence (n=10, x ± S)
Group | Dosage (μ g/kg) | Serum | Cardiac muscle homogenate |
NO (umol/L) | NO (umol/g albumen) | MDA (umol/g albumen) | Ca
2+(umol/g albumen)
|
The public Sodium Ferulate in 0.5%CMC JGS JGS JGS ground | - 25 50 100 150 | 73.45±6.31 76.29±6.59 1.65±11.64
△ 2.58±11.62
△ 77.95±11.36
| 7.19±1.16 8.49±1.8
△ 8.53±1.83
△ 9.03±1.52
△△ 8.95±2.27
△ | 0.1790.023 0.155±0.031
△ 0.153±0.022
△ 0.156±0.022
△ O.163±0.03
| 16.64±2.35 14.60±2.27
△ 13.91±3.48
△ 12.95±3.64
△△ 13.83±2.64
△ |
Compare with normal group:
△P<0.05,
△ △P<0.01, the t check
The result shows:
Compare with solvent control group (ig0.5%CMC solution 4ml/kg), the T ripple was raised and decreased heart rate (P<0.05-0.01) after JGS100 μ g/kg can significantly reduce the not normal incidence rate of injection of pituitrin, antagonism injection of pituitrin, JGS25,50 μ g/kg and positive control DIAOXINXUE KANG also have the antagonism ECG T wave to raise trend with decreased heart rate, but not statistically significant; JGS25,50 and 100 μ g/kg all can reduce serum CK level (P<0.05-0.01), the NO level of remarkable increase serum and myocardium homogenate (P<0.05-0.01), increase serum calcium ion concentration and reduce myocardium homogenate calcium ion that (P<0.05-0.01) concentration has alleviated calcium overload in the myocardial cell that injection of pituitrin causes effectively.JGS25,50 and 100 μ g/kg can also reduce myocardium homogenate lipid peroxidation product-MDA content (P<0.05), thereby have reduced the effect of radical pair induced myocardial injury.Positive control DIAOXINXUE KANG group also has above-mentioned variation, but free its effect of its nonreactive.See the above table 1-5.
Two, inducing the myocardial infarction experiment with the Chinese People's Anti-Japanese Military and Political College of the present invention Mus coronary ligation is example
90 of SD male rats, be divided into 6 groups at random, the basic, normal, high dosage group of JGS is igJGS extract 25,50,100 μ g/kg (in general flavone content) respectively, other establishes solvent control group ig 4ml/kg0.5%CMC solution, positive controls ig DIAOXINXUE KANG 150mg/kg (in the total Saponin of steroidal), and sham operated rats ig4ml/kg0.5%CMC solution (threading, not ligation).The above-mentioned all continuous gastric infusion of rat 5 days of respectively organizing, after irritating stomach in the 5th day 20 minutes, the anesthesia of lumbar injection 50mg/kg pentobarbital sodium, adopt Medlab5.5 bio signal acquisition system to trace one section II continuously and lead electrocardiogram, separate tracheal intubation behind the trachea, connect the animal artificial respirator and practice artificial respiration, during breathing than 1: 2, respiratory frequency 60 times/minute, tidal volume 3ml/100g.Open breast in second and third intercostal of left breast, expose heart, respectively at writing down one time electrocardiogram after the ligation in 60 minutes, 120 minutes, 180 minutes again.The rat broken end is got blood after 180 minutes, surveys serum CK-MB, Ca
2+, NO content; It is dirty to core, and separates left ventricle and weighs, and 0 ℃ is divided into the thick thin slice of 0.1cm with left ventricle after freezing, place 37 ℃ of 0.5%NBT liquid dyeing 15 minutes, 10% formalin fixed is distinguished myocardial infarction area and non-infarct, weigh respectively, calculate the percentage ratio that infarct accounts for left ventricular mass.
With electrocardiogram, serum CK-MB, Ca
2+, overall merit JGS such as biochemical indicator such as NO content and myocardial infarction percentage ratio Chinese People's Anti-Japanese Military and Political College's Mus myocardial infarction effect, the result shows:
Compare with solvent control group (ig0.5%CMC solution 4ml/kg), JGS25,50 and 100 μ g/kg all can significantly reduce the myocardium weight (P<0.05-0.01) of left chamber infarcted region behind the rat coronary ligation, increase the myocardium weight (P<0.05) of the non-infarcted region of rat left chamber, the ratio of left side chamber infarcted region reduces obviously that (P<0.05-0.01), positive controls DIAOXINXUE KANG group also has above-mentioned variation; Compare with sham operated rats, left chamber infarcted region of each dosage group rat of solvent control group, positive controls and JGS and left chamber infarcted region percentage ratio all have remarkable increase, see the following form 1.
The influence of the myocardial infarction district weight that table 1 JGS causes the rat coronary ligation (x ± S)
Group | Dosage (μ g/kg) | Number of animals (only) | Chamber, left side infarcted region cardiac muscle weight (g) | Chamber, a left side non-infarcted region cardiac muscle weight (g) | Left side chamber infarcted myocardium weight ratio (%) |
The sham-operation of 0.5%CMC JGS JGS JGS DIAOXINXUE KANG | 4ml/kg 25 50 100 150 4ml/kg | 14 10 10 10 14 14 | 0.215±0.045
△△ 0.150±0.041
**△△ 0.151±0.072
*△△ 0.133±0.071
**△△ 0.136±0.072
**△△ 0.032±0.052
** | 0.437±0.096
△△ 0.491±0.047
*△△ 0.501±0.083
*△△ 0.514±0.056
*△△ 0.505±0.082
△△ 0.616±0.102
** | 33.7±9.3
△△ 23.4±6.4
**△△ 23.3±11.0
*△△ 20.1±10.0
**△△ 21.2±11.5
**△△ 5.3±8.8
** |
Compare with 0.5%CMC solution group,
*P<0.05,
*P<0.01; Compare with sham operated rats,
△P<0.05,
△ △P<0.01 is the t check
Table 2 JGS to the rat coronary ligation after 3 hours serum NO level, CK-MB, Ca
2+Influence (x ± S, n=10)
Group | Dosage (μ g/kg) | NO (umol/L) | CK-MB (U/L) | Ca
2+ (mmol/L)
|
The sham-operation of 0.5%CMC JGS JGS JGS DIAOXINXUE KANG | 4ml/kg 25 50 100 150 4ml/kg | 26.1±7.0
△△ 10.8±8.8
** 17.7±8.2 21.4±18.5 51.9±54.0
△ 12.5±4.2
| 2217±2393 3018±2985 1911±1615 2836±3142 2207±1887
△ 1791±1677
| 2.33±0.11 2.33±0.062 2.25±0.074 2.32±0.088 2.32±0.13 2.33±0.090 |
Compare with 0.5%CMC solution group,
*P<0.05,
*P<0.01; Compare with sham operated rats,
△P<0.05,
△ △P<0.01 is the t check
The specific embodiment
Embodiment 1: get 500 kilograms of plant Herba Elsholtziae Blandae, after cleaning, extract total flavones by technological process shown in Figure 1, this extracting method has used the technology of using always on the pharmaceutics.
Embodiment 2: with extractive of general flavone and the adjuvant hydroxypropyl cellulose of implementing 1 gained, in extractive of general flavone: the ratio of adjuvant=99: 1, make the capsule of medicine of the present invention by technological process shown in Figure 2, this preparation technology has adopted technology commonly used on the pharmaceutics.
Embodiment 3: with extractive of general flavone and the adjuvant hydroxypropyl cellulose of implementing 1 gained, in extractive of general flavone: the ratio of adjuvant=65: 35, make the capsule of medicine of the present invention by technological process shown in Figure 2, this preparation technology has adopted technology commonly used on the pharmaceutics.
The technological process explanation
Be following several extracting mode in the technological process shown in Figure 1:
Scheme one:
Get the clean medical material of Herba Elsholtziae Blandae, it is an amount of to add water, distillation, and it is standby to collect volatile oil.
Scheme two:
Get the clean medical material of Herba Elsholtziae Blandae, after an amount of industrial alcohol or wetted with methanol, the dress percolator adds solvent and carries out percolation, collects percolate, reclaims to concentrate, and drying under reduced pressure gets dry extract, and pulverizes the back and mixes with adjuvant, granulates, and encapsulated, blister package, outer package are promptly.
Scheme three:
Get the clean medical material of Herba Elsholtziae Blandae, it is an amount of to add water, decocts, and filters, and concentrates the back with the alcohol precipitation, gets supernatant, reclaims to concentrate, and drying under reduced pressure gets dry extract, and pulverizes the back and mixes with adjuvant, granulates, and encapsulated, blister package, outer package are promptly.
Scheme four:
Get the clean medical material of Herba Elsholtziae Blandae, processing industry ethanol or methanol are an amount of, and reflux, extract, reclaims concentrated back and uses water precipitation, gets supernatant, reclaim to concentrate, and drying under reduced pressure gets dry extract, and mixes with adjuvant after pulverizing, and granulates, and encapsulated, blister package, outer package are promptly.
Above-mentioned four kinds of schemes can be carried out simultaneously, also can carry out separately.