The specific embodiment:
The preparation of bamboo extract:
Preparation example 1: alkaline extraction
Adopt down Folium Bambusae or bamboo branch, tap water cleans two times, and oven dry is pulverized, claim 500g to add the 8000ml saturated limewater and soaked 1 hour, heating 1h (80~100 ℃), filtered while hot, residue adds 5000ml saturated limewater reheat and extracts 1h, filter, concentrate, pulverize, be sealed in the sample bottle standby.Obtain alkali and carry matter sample one 80g.
Preparation example 2: alkaline extraction
Adopt down Folium Bambusae or bamboo branch, tap water cleans two times, and oven dry is pulverized, claiming 500g to add 8000ml PH is 3 sodium hydroxide ethanol (concentration of alcohol is 50%) solution soaking 1 hour, heating 1h (80~100 ℃), filtered while hot, residue adds 5000ml sodium hydroxide alcoholic solution reheat and extracts 1h, filter, concentrate, pulverize, be sealed in the sample bottle standby.Obtain alkali and carry matter sample two 80g.
Preparation example 3: alcohol extracting method
Adopt down Folium Bambusae or bamboo branch, tap water cleans two times, and oven dry is pulverized, claim that 500g adds 8000ml 50% alcohol solution dipping 1 hour, heating 1h (80~100 ℃), filtered while hot, residue adds 5000ml 50% alcoholic solution reheat and extracts 1h, filter, concentrate, pulverize, be sealed in the sample bottle standby.Obtain ethanol extract sample one 75g.
Preparation example 4: alcohol extracting method
Adopt down Folium Bambusae or bamboo branch, tap water cleans two times, and oven dry is pulverized, and claims that 500g adds 8000ml 30% methanol solution, soaked 1 hour, heating 1h (80~100 ℃), filtered while hot, residue adds 5000ml 30% methanol solution reheat and extracts 1h, filter, concentrate, pulverize, be sealed in the sample bottle standby.Obtain ethanol extract sample two 77g.
Carry out pharmacological testing with above-mentioned sample:
Experimental technique:
45Ca strides the film measuring method
Material
The Wistar rat, body weight 200+20g, male and female dual-purpose.
45CaCl
2Solution is Britain AmerSham company product, and specific radioactivity 1570MBq/gCa, other medicine are commercially available analytical pure.
Experiment liquid
PSS liquid (m mol/L): NaCl 137, CaCl
21.5, MgCl
21.0, KCl 3.0, HEPES20, C
6H
12O
6H
2O
10, pH=7.4; High potassium PSS liquid (m mol/L): KCl 100, NaCl 40 all the other with PSS liquid, pH=7.4; EGTA liquid (m mol/L): EGTA l0 does not contain CaCl
2, all the other same PSS; The toluene scintillation solution: PPO 5.0g, POPOP 0.4g, ethylene glycol 300ml, naphthalene 60g are settled to 1000ml with toluene.
45Ca strides flow measurement step in the film:
Press document: Williams GT.Cell, 1991,65:1097 and Zhang Tingjie angiocardiology progress, 1995,16 (1): 43 method is also improved.Hit dizzy rat and take out thoracic aorta, the heart, liver, lung, kidney ... Deng, place PSS solution to remove connective tissue gently, be cut into small pieces, place 37 ℃ of insulations of PSS liquid 1h, immigration contains
45CaPSS solution equilibria 10min.Tremulous pulse bar or continue to stay in this solution, or move in norepinephrine (NE) solution or move into and contain
45The high potassium liquid of Ca is handled 10min.Above step all is at 37 ℃ and logical O
2Carry out under the condition.To organize taking-up again, place 0-2 ℃ of no calcium EGTA solution washing 1h, and put into respectively after blotting arterial ring and weigh with filter paper then and measure bottle and add 20%HClO
425 μ L and 30%H
2O
250 μ L digest 20min in 80 ℃ of water-baths, radioactivity dpm is measured after adding the dark 3h of placement of toluene scintillation solution 4ml in the cooling back in liquid scintillation instrument.Ca
2+Interior flow is calculated by following formula:
Measurement data is represented with x ± S.D, and is carried out the t check.
45Ca strides the excessive measuring method of film:
Getting rat tissue's cell → 37 ℃ PSS liquid hatches → contains
45Ca enters ℃ EGTA flush away extracellular, cell → 4 in a large number by the PDC calcium channel
45Make among ℃ PSS of Ca → 37
45Ca is excessive → regularly surveys among the PSS
45The cpm of Ca is residue in excessive speed → survey cell
45Ca →
45The interior flow of Ca subtracts remaining quantity and promptly gets excessive amount.
In excessive process, select PSS liquid, PSS+ epinephrine, the high potassium of PSS for use, represent respectively that promptly stationary stream calcium channel (Leak), receptor calcium channel (ROC) and voltage rely on the excessive of calcium channel (PDC).
Experimental result
Preparation technology's difference, pharmacological action is variant
The ethanol extraction of Folium Bambusae or bamboo branch and alkaline process extract rely on calcium channel (PDC) to rat aorta voltage
45The influence of stream sees Table I, Table II in the Ca.
Table I Folium Bambusae ethanol extract is to PDC
45The influence (n=8) of stream in the Ca
Classification | Concentration (mg/ml) | Alcohol extraction sample one | Alcohol extraction sample two |
Contrast ethanol extract contrast ethanol extract | 0 0.02 0.10 0.50 | 193.18±10.48 185.22±24.34 187.92±5.67 182.86±6.47 | 193.18±10.48 188.92±9.14 158.52±8.01 143.56±11.41 |
Table II Folium Bambusae alkali is carried thing to PDC
45The influence (n=8) of stream in the Ca
Classification | Concentration (mg/ml) | The alkali product one that get sample | The alkali product two that get sample |
Contrast alkali is carried thing contrast alkali and is carried thing | 0 0.02 0.10 0.50 | 185.87±17.13 147.37±15.72 157.37±14.24 128.91±15.92 | 185.87±17.13 160.37±16.06 154.76±13.36 106.20±9.76 |
The result of comparison sheet I, Table II finds out, sample one and contrast zero difference, sample two in the ethanol extract
45The stream effect is better than sample one in the Ca.Carry at alkali that the two all is to have shown good effect in the thing.Relatively alkali is put forward the effect of thing and ethanol extract effect, and alkali is carried thing retardance PDC calcium channel, reduces
45The effect of stream is better than ethanol extract in the Ca.Retardation under 0.02mg/ml concentration, just occurs, and the Concentraton gradient effect is arranged.
Bamboo extract, Folium Ginkgo flavone, Verapamil have similar action rule to calcium channel
Bamboo, Folium Ginkgo, Verapamil etc. see Table III to the action characteristic of the rat heart, liver, lung, kidney PDC calcium channel.
The Table III bamboo extract is to the influence (n=8) of internal organs organ PDC calcium channel
Classification | Concentration (mg/ml) | Tremulous pulse | The heart | Liver | Lung | Kidney |
Contrast | 0 | 199.32±12.26 | 753.16±63.63 | 425.07±60.64 | 357.86±44.40 | 438.28±63.90 |
Bamboo alkali is carried thing | 0.5 | 135.78±10.87 | 528.02±24.82 | 413.69±49.24 | 223.36±46.46 | 351.08±36.37 |
The bamboo ethanol extract | 0.5 | 150.27±5.61 | 549.80±50.30 | 378.35±38.54 | 251.36±23.38 | 330.24±23.80 |
Folium Ginkgo | 0.5 | 146.96±22.35 | 642.30±58.1 | 360.62±32.98 | 285.43±12.84 | 371.58±22.65 |
Verapamil | 0.1 | 110 | 589.02±49.88 | 310.27±15.52 | 319.11±11.43 | 407.94±17.80 |
Can find out that from last table bamboo extract all has regulating and controlling effect to the rat aorta heart, liver, lung, kidney.When concentration is 0.5mg/ml, tremulous pulse, the heart, lung action effect are had identical rule with Folium Ginkgo, verapamil, because of verapamil is chemical medicine, purity is higher, drug action under low concentration very just clearly.
Bamboo extract is to the internal organs organ
45The excessive influence of Ca sees Table IV
The Table IV Folium Bambosae extract is right
45The influence (n=87) that Ca is excessive
Classification | Concentration | Tremulous pulse | The heart | Liver | Lung | Kidney |
Contrast | 0 | 57.08±1.19 | 316.40±22.46 | 211.45±4.71 | 165.80±10.90 | 331.40±11.80 |
Folium Bambusae alkali is carried thing | 0.4 | 36.60±3.76 | 271.53±28.28 | 220.46±29.82 | 160.56±16.11 | 340.18±32.65 |
Bamboo skill alkali is carried thing | 0.4 | 41.70±4.7 | 285.16±30.60 | 174.42±25.22 | 154.69±30.40 | 262.86±33.61 |
Folium Ginkgo | 0.4 | 42.90±9.51 | 271.45±42.51 | 185.89±25.96 | 142.11±25.33 | 239.89±28.80 |
As seen from Table 4, Folium Bambusae, bamboo skill, Folium Ginkgo, alkali are carried thing can impel Ca in the heart, liver, lung, kidney, the arterial cell under 0.4mg/ml concentration
2+To excessive, to reduce Ca in the cell
2+The injury that the overload pair cell causes.Wherein relatively consistent to tremulous pulse, heart action rule, to lung, liver, kidney not clearly.
From above-mentioned experiment as can be seen, bamboo extract both can promote unnecessary Ca in the cell
2+Excessive, can stop extracellular Ca again
2+Interior stream, it is to multiple organ cell's film calcium channel regulating action, be of value to the stability of keeping intracellular calcium concentration, thereby to cardiovascular and cerebrovascular disease have the treatment and health care function, drug action and Folium Ginkgo have comparability, except that can raw material as preparation CCB (calcium channel blocker) medicine, can also be to keeping normal Ca in the cell
2+Concentration plays regulating action.Normal Ca in the cell
2+The destruction of stability of concentration is the early stage and key condition that cell is injured.A large amount of calcium and pigmentation are generally all arranged in the senile cell; Ca in aging dysmnesia and the brain cell synapse
2+Serious overload is relevant; Body is inflamed or lacks O
2All with position, place cell Ca
2+Concentration raises.Intracellular calcium is unbalance to be considered to " co-channel " of the cell injury that a variety of causes causes.Thereby extract of the present invention makes cell recover normal function again for getting rid of unnecessary calcium in the cell, provides new way for preventing and treating multiple diseases such as aging, cancer.
The invention will be further described below by embodiment, but be not limitation of the present invention.
Embodiment one
The preparation of bamboo extract:
Adopt down Folium Bambusae or bamboo branch, tap water cleans two times, and oven dry is pulverized, claim 500g to add 8000ml saturated limewater (pH14) and soaked 1 hour, heating 1h (80~100 ℃), filtered while hot, residue adds 5000ml saturated limewater reheat and extracts 1h, filter, concentrate, pulverize, be sealed in the sample bottle standby.Obtain extract 80g.
Embodiment two
The preparation of bamboo extract:
Folium Bambusae or bamboo branch are cut into the 1-3cm segment with chopper, dry or directly extract, add 10% sodium bicarbonate (containing 1% Borax) by solid-to-liquid ratio at 1: 16 with the aquatic foods product, soak 2h and do not have heated and boiled 1h, filtered while hot, residue is by adding sodium bicarbonate (containing 1% Borax) at 1: 8, boiled 1 hour again, and filtered, twice filtrate merges, concentrate, oven dry is pulverized, and it is standby to bottle.Obtaining extract is 18% of raw material (dry product).
With the pharmacological experimental method of front, measure it to retardance extracellular Ca
2+Ca in interior stream and the promotion cell
2+Excessive influence, unanimity as a result.
Embodiment three
The preparation of bamboo extract:
Adopt down Folium Bambusae or bamboo branch, tap water cleans two times, oven dry, chopping, title 500g adds alcoholic solution (pH10 contains alcohol 30%) the immersion half an hour that 8000ml contains sodium hydroxide, heating 1h (80~100 ℃), filtered while hot, residue adds the above-mentioned alkali alcohol of 5000ml liquid reheat and extracts 1h, filters, and concentrates, pulverize, be sealed in the sample bottle standby.Obtain extract 74g.
With the pharmacological experimental method of front, measure it to retardance extracellular Ca
2+Ca in interior stream and the promotion cell
2+Excessive influence, unanimity as a result.
Embodiment four
Get the bamboo extract 100g of embodiment one or two or three method preparation, add starch, each 10g of dextrin, mixing is granulated, drying, and granulate in the special-purpose bag of the bagged steeping drug of packing into, is teabag.
Embodiment five
Get the bamboo extract 100g of embodiment one or two or three method preparation, add starch, each 10g of dextrin, mixing is granulated, drying, and granulate in the hard capsule of packing into, is capsule.
Embodiment six
Get the bamboo extract 100g of embodiment one or two or three method preparation, add starch, each 10g of dextrin, mixing is granulated, drying, and granulate adds magnesium stearate 2g, and tabletting is conventional tablet.
Embodiment seven
Get the bamboo extract 100g of embodiment one or two or three method preparation, add smart in a subtle way cellulose 40g, cross-linked pvp 40g, lactose 10g, mixing is granulated, drying, and granulate adds magnesium stearate 2g, and tabletting is dispersible tablet.
Embodiment eight
Get the bamboo extract 100g of embodiment one or two or three method preparation, add starch, each 10g of dextrin or do not add, general system micropill, drying, the bag extended release coatings is slow releasing pill.This ball incapsulates, and is slow releasing capsule.
Embodiment nine
Get the bamboo extract 100g of embodiment one or two or three method preparation, add hydroxypropyl methylcellulose 20g, ethyl cellulose 40g etc., mixing is granulated, and is sustained-release granular formulation.This granule incapsulates, and is slow releasing capsule.This granule tabletting is slow releasing tablet.
Embodiment ten
Get the bamboo extract 100g of embodiment one or two or three method preparation, add hawthorn extract 30g or Fructus Crataegi micropowder 50g, protein sugar 0.3g, be dissolved in or be suspended in the aqueous solution, be health beverage.