Clear stone Chinese medicine composition and preparation method thereof
Technical field
The invention belongs to application in TCM technical field, is related to a kind of clear stone Chinese medicine composition and preparation method thereof.
Background technology
Urolithiasis is urinary system common disease and frequently-occurring disease, wherein kidney stone incidence of disease highest, and 80% is calcinm oxalate calculus.
Traditional Chinese medicine thinks:Kidney stone belongs to the category of the illness such as " urolithiasis ", " sand leaching ", " blood strangury ", " pain in the back ".It is clinically to show as urinating more
Calcium, oxalic acid, the discharge such as phosphate discharge increase, magnesium ion, citric acid, citric acid reduce, and are often accompanied by blood calcium rise in liquid.Kidney
Calculus pathogenic factor and mechanism are more complicated, are the coefficient results such as inherent cause and environmental factor.Research is thought at present,
The supersaturation of calcium oxalate is the prerequisite that kidney stone is formed, then a system such as experience nucleation, growth, aggregation and solid phase transformation
Row physical and chemical process.Therefore, a kind of medicine of effectively treatment kidney stone is developed, is the problem currently explored.
The content of the invention
It is an object of the invention to solve at least the above and/or defect, and provide at least will be described later it is excellent
Point.
It is a still further object of the present invention to provide a kind of clear stone Chinese medicine composition.
It is of the invention separately to there is a purpose to be to provide a kind of preparation method of clear stone Chinese medicine composition.
Therefore, technical scheme provided by the invention is:
A kind of clear stone Chinese medicine composition, including:
Lygodium japonicum 200-1600 parts and pyrrosia lingua 150-1200 parts.
Preferably, described clear stone Chinese medicine composition, the component of following parts by weight is included:
Bark of official magnolia 150-1200 parts, Poria cocos 200-1600 Fen , pivots store 150-1200 parts, oriental wormwood 150-1200 parts, rhizoma alismatis 200-
1600 parts, fringed pink 150-1200 parts and rhizoma imperatae 150-1200 parts.
Preferably, described clear stone Chinese medicine composition, the component of following parts by weight is included:
Shell of areca nut 200-1600 parts, Radix Astragali 150-1200 parts, root of three-nerved spicebush 200-1600 parts and honey-fried licorice root 100-800 parts.
Preferably, in described clear stone Chinese medicine composition, the clear stone Chinese medicine composition includes following parts by weight
Component:800 parts of the Lygodium japonicum, 600 parts of pyrrosia lingua, 600 parts of the bark of official magnolia, 600 parts of the Fen , pivots of Poria cocos 800 storage, 600 parts of oriental wormwood, rhizoma alismatis
800 parts, 600 parts of fringed pink, 600 parts of rhizoma imperatae, 800 parts of the shell of areca nut, 600 parts of the Radix Astragali, 400 parts of 800 parts of the root of three-nerved spicebush and honey-fried licorice root.
Preferably, in described clear stone Chinese medicine composition, the clear stone Chinese medicine composition includes the group of following parts by weight
Point:
1600 parts of the Lygodium japonicum, 1200 parts of pyrrosia lingua, 1200 parts of the bark of official magnolia, the Fen , pivots of Poria cocos 1600 store 1200 parts, oriental wormwood 1200
Part, 1600 parts of rhizoma alismatis, 1200 parts of fringed pink, 1200 parts of rhizoma imperatae, 1600 parts of the shell of areca nut, 1200 parts of the Radix Astragali 1600 parts of the root of three-nerved spicebush and processs
800 parts of radix glycyrrhizae.
Preferably, in described clear stone Chinese medicine composition, the clear stone Chinese medicine composition includes following parts by weight
Component:Lygodium japonicum 400-800 parts, pyrrosia lingua 300-600 parts, bark of official magnolia 300-600 parts, Poria cocos 400-800 Fen , pivots store 300-600 parts,
Oriental wormwood 300-600 parts, rhizoma alismatis 400-800 parts, fringed pink 300-600 parts, rhizoma imperatae 300-600 parts, shell of areca nut 400-800 parts are yellow
Stilbene 300-600 parts, root of three-nerved spicebush 400-800 parts and honey-fried licorice root 200-400 parts.
Preferably, in described clear stone Chinese medicine composition, the clear stone Chinese medicine composition includes following parts by weight
Component:400 parts of Lygodium japonicum, 300 parts of pyrrosia lingua, 300 parts of the bark of official magnolia, 300 parts of the Fen , pivots of Poria cocos 400 storage, 300 parts of oriental wormwood, 400 parts of rhizoma alismatis,
200 parts of 300 parts of fringed pink, 300 parts of rhizoma imperatae, 400 parts of the shell of areca nut, 300 parts of the Radix Astragali, 400 parts of the root of three-nerved spicebush and honey-fried licorice root.
Preferably, in described clear stone Chinese medicine composition, the clear stone Chinese medicine composition is granular preparation, mixture, glue
Capsule preparation, pill or tablet, the pill includes extraction medicinal extract pill and former powder is used as medicine pill.
A kind of preparation method of clear stone Chinese medicine composition, comprises the following steps:
Step 1: in terms of parts by weight, the bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis are taken, alcohol reflux is added and extracts 2 hours, filtration, filtrate
Recovery ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25;
Step 2: in terms of parts by weight, the dregs of a decoction and Lygodium japonicum, pyrrosia lingua, Poria cocos, pivots storage, the mattress that are filtrated to get in step 1
Old, fringed pink, rhizoma imperatae, the shell of areca nut, the Radix Astragali and honey-fried licorice root mixing, are soaked 2 hours, decoction is secondary, 2 hours every time, decocting liquid
Filtration, merges, and concentrate with the filtrate in step 1, lets cool afterwards, and then plus ethanol makes alcohol content stand, filter up to 60%,
Filtrate recycling ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25, with the above-mentioned bark of official magnolia, crow
Medicine and rhizoma alismatis medicinal extract, which merge, to be mixed, and obtains clear stone Chinese medicine composition;
Wherein, described Lygodium japonicum 200-1600 parts, pyrrosia lingua 150-1200 parts, bark of official magnolia 150-1200 parts, Poria cocos 200-1600
Fen , pivots store 150-1200 parts, oriental wormwood 150-1200 parts, rhizoma alismatis 200-1600 parts, fringed pink 150-1200 parts, rhizoma imperatae 150-1200
Part, shell of areca nut 200-1600 parts, Radix Astragali 150-1200 parts, root of three-nerved spicebush 200-1600 parts and honey-fried licorice root 100-800 parts.
Preferably, in the preparation method of described clear stone Chinese medicine composition, also comprise the following steps:
Step 3: adding dextrin and flavouring into the clear stone Chinese medicine composition, mix, pelletize, dry, particle is made
1000 parts, obtain clear stone particle.
Preferably, in the preparation method of described clear stone Chinese medicine composition, in the step 1, the matter of the ethanol of addition
Amount fraction is 40-80%, and addition is 5-10 times of foregoing Chinese medicine weight;
In the step 2, the mass fraction of the ethanol of addition is 50-95%.
Preferably, in the preparation method of described clear stone Chinese medicine composition, in the step 2, before the water of addition is
State Chinese medicine weight 5-10 times.
The present invention comprises at least following beneficial effect:
The clear stone Chinese medicine composition of the present invention is by the taste traditional Chinese medicine extraction of pyrrosia lingua, Lygodium japonicum, oriental wormwood, the bark of official magnolia, rhizoma alismatis etc. 13 essence
System forms, and has the effect of niter is treating stranguria, cooling blood and relieving pain.In said preparation prescription, Lygodium japonicum inducing diuresis for treating strangurtia analgesic, pyrrosia lingua diuresis is led to
Leaching, and energy cooling blood and hemostasis, Lygodium japonicum obtain pyrrosia lingua niter and can stopped blooding, for face lithiasis often with the disease of blood urine, the two niter is treating stranguria,
Cooling blood and relieving pain, it is altogether monarch drug in a prescription.Pivot stores inducing diuresis for treating strangurtia and has heat-clearing, oriental wormwood clearing heat and promoting diuresis concurrently, and rhizoma alismatis eliminating dampness and diuresis purges heat, three
Make damp and hot from urine to go out;Bark of official magnolia eliminating dampness promoting the circulation of qi, Poria cocos inducing diuresis with bland drug and can invigoration spleen qi, so as to strengthen dampness removing function, fringed pink,
Imperata rhizome heat-clearing diuretic, cooling blood and hemostasis, pyrrosia lingua Yu pivots storage, oriental wormwood, the work(of rhizoma alismatis are helped, is altogether ministerial drug.The shell of areca nut, Radix Astragali promoting the circulation of qi profit
Urine detumescence, root of three-nerved spicebush promoting qi circulation and relieving pain is adjutant, and three's promoting the circulation of qi is helped below stone, arranges stone and goes out.Radix glycyrrhizae coordinating the drug actions of a prescription, relieving spasm to stop pain are simultaneous
Energy heat-clearing, is adjuvant.
Test result indicates that calcium, oxalic acid content significantly reduce in clear each treatment group's nephridial tissue of stone particle, pathological examination result
It has been shown that, in kidney calcium oxalate crystal distribution reduce, tubular ectasia, come off, necrosis is obviously improved.Clear stone particle is prompted effectively to press down
The formation and promotion calculus discharge of calcinm oxalate calculus processed.Meanwhile urinate Analysis of Biochemical result and show that clear stone particle can be carried significantly
High 24h urine volume, promote urine uric acid excretion, alkalized urine.Increase urine volume and alkalized urine can reduce calcium oxalate supersaturation and separate out, and add
By force to the souring of urinary tract, calcium oxalate crystals are reduced to the adhesive attraction of renal cells, suppress the formation of calculus.Urine
Acid can be combined with calcium oxalate crystals, promote the growth of calcium oxalate crystals.Clear stone particle urinates the excretion of uric acid by increasing, and reduces urine
Acid reduces the formation of calcium oxalate crystal to the facilitation of calculus.
Clear stone particle can significantly reduce serum calcium ion, creatinine, urea content, alleviate impaired renal function symptom, improve kidney
Bead filtering function, protect renal function.The oxalic acid of high concentration can cause renal cells to produce reactive oxygen species, cause thin
Born of the same parents' apoptosis, necrosis and cell membrane surface characteristic changing, promote calculus.Clear stone particle can improve polyphenoils in nephridial tissue
SOD level, promote oxalic acid excretion, the damage of renal cells is reduced, so as to suppress the formation of calcium oxalate crystals.
In summary, clear stone particle can effectively reduce calcium in nephridial tissue, oxalic acid content, protect renal cells, promote
Enter the excretion of uric acid, alkalized urine, increase urine discharge capacity, so as to suppress the formation of calcinm oxalate calculus.
Meanwhile the present invention combines 1% ethylene glycol using 1% ammonium chloride and replicates kidney calcinm oxalate calculus model.Experimental result table
Bright, model control group experimental rat serum calcium, creatinine, urea significantly increase, in nephridial tissue oxalic acid, calcium content and pathological examination with
Blank group has significant difference.Represent the success of Rat renal calcinm oxalate calculus model copy.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Brief description of the drawings
Fig. 1 is the absorption spectrum curve of catalysing tube and on-catalytic pipe oxalic acid in the one of embodiment of the present invention;
Fig. 2 is the one of embodiment mesoxalic acid canonical plotting of the present invention;
Fig. 3 is calcium ion canonical plotting in the one of embodiment of the present invention;
Fig. 4 is blank control group HE Coloration experiment Rat renals under 400 times of multiplication factors in the one of embodiment of the present invention
Dirty tissue micro-image;
Fig. 5 is 400 times of multiplication factor drag control group HE Coloration experiment Rat renals in the one of embodiment of the present invention
Dirty tissue micro-image;
Fig. 6 is Shenshitong group HE Coloration experiment rat kidney under 400 times of multiplication factors in the one of embodiment of the present invention
Organize micro-image;
Fig. 7 is clear stone particle low dose group HE Coloration experiments under 400 times of multiplication factors in the one of embodiment of the present invention
Rat kidney tissue micro-image;
Fig. 8 is clear stone particle middle dose group HE Coloration experiments under 400 times of multiplication factors in the one of embodiment of the present invention
Rat kidney tissue micro-image;
Fig. 9 is clear stone particle high dose group HE Coloration experiments under 400 times of multiplication factors in the one of embodiment of the present invention
Rat kidney tissue micro-image;
Figure 10 is clear influence block diagram of the stone particle to experimental rat body weight in the one of embodiment of the present invention;
Figure 11 is clear influence block diagram of the stone particle to experimental rat blood calcium in the one of embodiment of the present invention;
Figure 12 is clear influence block diagram of the stone particle to experimental rat creatinine in the one of embodiment of the present invention;
Figure 13 is clear influence block diagram of the stone particle to experimental rat urea in the one of embodiment of the present invention;
Figure 14 is clear influence block diagram of the stone particle to experimental rat urine volume in the one of embodiment of the present invention;
Figure 15 is the influence block diagram that clear stone particle urinates experimental rat pH value in the one of embodiment of the present invention;
Figure 16 is clear influence block diagram of the stone particle to experimental rat urinary calcium in the one of embodiment of the present invention;
Figure 17 is the influence block diagram that clear stone particle urinates experimental rat uric acid in the one of embodiment of the present invention;
Figure 18 is the influence block diagram that clear stone particle urinates experimental rat oxalic acid in the one of embodiment of the present invention;
Influence block diagrams of the Figure 19 for clear stone particle in the one of embodiment of the present invention to experimental rat kidney calcium;
Influence block diagrams of the Figure 20 for clear stone particle in the one of embodiment of the present invention to experimental rat kidney oxalic acid;
Figure 21 is clear influence block diagram of the stone particle to experimental rat SOD in the one of embodiment of the present invention;
Figure 22 is clear influence block diagram of the stone particle to experimental rat Kidney coefficients in the one of embodiment of the present invention.
Embodiment
The present invention is described in further detail below in conjunction with the accompanying drawings, to make those skilled in the art with reference to specification text
Word can be implemented according to this.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein do not allot one or more
The presence or addition of individual other elements or its combination.
The present invention provides a kind of clear stone Chinese medicine composition, includes the component of following parts by weight:
Lygodium japonicum 200-1600 parts and pyrrosia lingua 150-1200 parts.
In such scheme, preferably, the clear stone Chinese medicine composition, include the component of following parts by weight:
Bark of official magnolia 150-1200 parts, Poria cocos 200-1600 Fen , pivots store 150-1200 parts, oriental wormwood 150-1200 parts, rhizoma alismatis 200-
1600 parts, fringed pink 150-1200 parts and rhizoma imperatae 150-1200 parts.
In such scheme, preferably, the clear stone Chinese medicine composition, include the component of following parts by weight:
200-1600 parts, Radix Astragali 150-1200 parts, root of three-nerved spicebush 200-1600 parts and honey-fried licorice root 100-800.
In a preferred embodiment, the clear stone Chinese medicine composition includes the component of following parts by weight:
800 parts of the Lygodium japonicum, 600 parts of pyrrosia lingua, 600 parts of the bark of official magnolia, the Fen , pivots of Poria cocos 800 store 600 parts, and 600 parts of oriental wormwood is damp
Rush down 800 parts, 600 parts of fringed pink, 600 parts of rhizoma imperatae, 800 parts of the shell of areca nut, 600 parts of the Radix Astragali, 400 parts of 800 parts of the root of three-nerved spicebush and honey-fried licorice root.
In a preferred embodiment, the clear stone Chinese medicine composition includes the component of following parts by weight:
1600 parts of the Lygodium japonicum, 1200 parts of pyrrosia lingua, 1200 parts of the bark of official magnolia, the Fen , pivots of Poria cocos 1600 store 1200 parts, oriental wormwood 1200
Part, 1600 parts of rhizoma alismatis, 1200 parts of fringed pink, 1200 parts of rhizoma imperatae, 1600 parts of the shell of areca nut, 1200 parts of the Radix Astragali 1600 parts of the root of three-nerved spicebush and processs
800 parts of radix glycyrrhizae.
In some preferred embodiments, the clear stone Chinese medicine composition includes the component of following parts by weight:Lygodium japonicum
400-800 parts, pyrrosia lingua 300-600 parts, bark of official magnolia 300-600 parts, Poria cocos 400-800 Fen , pivots store 300-600 parts, oriental wormwood 300-600
Part, rhizoma alismatis 400-800 parts, fringed pink 300-600 parts, rhizoma imperatae 300-600 parts, shell of areca nut 400-800 parts, Radix Astragali 300-600 parts,
Root of three-nerved spicebush 400-800 parts and honey-fried licorice root 200-400 parts.
In a preferred embodiment, the clear stone Chinese medicine composition includes the component of following parts by weight:Lygodium japonicum
400 parts, 300 parts of pyrrosia lingua, 300 parts of the bark of official magnolia, 300 parts of the Fen , pivots of Poria cocos 400 storage, 300 parts of oriental wormwood, 400 parts of rhizoma alismatis, 300 parts of fringed pink,
200 parts of 300 parts of rhizoma imperatae, 400 parts of the shell of areca nut, 300 parts of the Radix Astragali, 400 parts of the root of three-nerved spicebush and honey-fried licorice root.
In some preferred embodiments, the clear stone Chinese medicine composition be granular preparation, mixture, capsule preparations, pill or
Tablet, the pill includes extraction medicinal extract pill and former powder is used as medicine pill.Can with sugaring can not also sugaring, particle system is made
Agent.
The present invention also provides a kind of preparation method of clear stone Chinese medicine composition, comprises the following steps:
Step 1: in terms of parts by weight, the bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis are taken, alcohol reflux is added and extracts 2 hours, filtration, filtrate
Recovery ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25;
Step 2: in terms of parts by weight, the dregs of a decoction and Lygodium japonicum, pyrrosia lingua, Poria cocos, pivots storage, the mattress that are filtrated to get in step 1
Old, fringed pink, rhizoma imperatae, the shell of areca nut, the Radix Astragali and honey-fried licorice root mixing, are soaked 2 hours, decoction is secondary, 2 hours every time, decocting liquid
Filtration, merges, and concentrate with the filtrate in step 1, lets cool afterwards, and then plus ethanol makes alcohol content stand, filter up to 60%,
Filtrate recycling ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25, with the above-mentioned bark of official magnolia, crow
Medicine and rhizoma alismatis medicinal extract, which merge, to be mixed, and obtains clear stone Chinese medicine composition;
Wherein, described Lygodium japonicum 200-1600 parts, pyrrosia lingua 150-1200 parts, bark of official magnolia 150-1200 parts, Poria cocos 200-1600
Fen , pivots store 150-1200 parts, oriental wormwood 150-1200 parts, rhizoma alismatis 200-1600 parts, fringed pink 150-1200 parts, rhizoma imperatae 150-1200
Part, shell of areca nut 200-1600 parts, Radix Astragali 150-1200 parts, root of three-nerved spicebush 200-1600 parts and honey-fried licorice root 100-800 parts.
In a preferred embodiment, the preparation method of the clear stone Chinese medicine composition also comprises the following steps:
Step 3: adding dextrin and flavouring into the clear stone Chinese medicine composition, mix, pelletize, dry, particle is made
1000 parts, obtain clear stone particle.
In a preferred embodiment, in the step 1, the mass fraction of the ethanol of addition is 40-80%, is added
Measure as 5-10 times of foregoing Chinese medicine weight;
In the step 2, the mass fraction of the ethanol of addition is 50-95%.
In a preferred embodiment, in the step 2, the water of addition is 5-10 times of foregoing Chinese medicine weight.
To make those skilled in the art more fully understand the present invention, examples below is now provided and illustrated.
Embodiment 1
【Effect experiment】
1.1 experiment material
1.1.1 experimental animal
SD rats:Cleaning grade, body weight 180-200g, male, health, the long-living Bioisystech Co., Ltd in Liaoning is purchased from, closed
Lattice card number:SCXK (the Liao Dynasty) 2015-0001.
1.1.2 medicine and reagent
Wherein, the prescription of clear stone particle is as follows:
【Low dosage prescription】
【Middle dosage prescription】
【High dose prescription】
1.1.3 key instrument
1.2 experimental method
1.2.1 the foundation and packet of rat model
Healthy male SD rat 48, body weight 180-200g, after adaptability is fed 1 week, sky is divided into using random number method
White control group, model control group, Shenshitong Zu, Cheongju stone particle low dose group, middle dose group, high dose group, every group 8.Except sky
White remaining outer each group of control group gives 1% ethylene glycol free water, 1% ammonium chloride 2mL/d gavages.After modeling 1 week, Shenshitong
Group, Cheongju stone particles low dose group, middle dose group, high dose group are respectively according to 2.7g/kg, 2.25g/kg, 4.5g/kg, 9.0g/kg
Gastric infusion, 2 times/day.Model control group, blank control group give the distilled water gavage of equivalent, give modeling while administration
Medicine, continuous gavage 3 weeks.
1.2.2 the collection and detection of sample
1.2.2.1 collection and the index analysis of urine specimen:Experiment terminates the previous day collection rat 24h urine and (added anti-
Rotten agent), for determining 24h urine volume, urine pH value, urine oxalic acid, urine Ca2+.Automatic biochemical analyzer measure urine Ca2+, colorimetric method for determining
Urinate oxalic acid.
1.2.2.2 collection and the index analysis of serum specimen:After modeling 4 weeks, the chloraldurate (matter of rats by intraperitoneal injection 10%
Measure fraction), after of flaccid muscles, abdominal vein punctures and takes blood about 5ml, injects centrifuge tube, stands 30 minutes, 3000r/min from
Heart 15min, extracts serum specimen, and -20 DEG C of preservations are to be measured.
1.2.2.3 the collection of kidney sample and index analysis:After rat extracting blood, double kidneys are completely cut immediately, separate kidney quilt
Film, normal saline flushing is wiped dry, weighs, left kidney be put into 10% neutral formalin (volume fraction) solution it is fixed, for doing HE dyes
Color is cut into slices, and the formational situation and Pathological of optical microphotograph Microscopic observation calcium oxalate crystal change;Right kidney is put into -70 DEG C of low temperature
Preserved in refrigerator, for determining Ca in Kidney coefficients, nephridial tissue2+, SOD and oxalic acid content.
1.2.2.4 Indexs measure analysis method
1. general status is observed:Observe the change of rat body weight, mental status, amount of drinking water, food-intake and mode of appearance
Etc. index, and record at any time.
2. blood biochemistry detects:With automatic clinical chemistry analyzer measure serum urea, creatinine, Ca2+It is horizontal.
3. urinate biochemistry detection:Urine Ca is determined with automatic clinical chemistry analyzer2+, uric acid.
4. renal tissue and oxalate crystal growth in healthy urine content:Using catalytic spectrophotometric nephridial tissue and oxalate crystal growth in healthy urine content.
5. Ca in renal tissue2+Content:Using methyl thymol blue (MTB) colorimetric method for determining Ca2+Content.
6. SOD contents in renal tissue:SOD kits (WST-1 methods).
7. Pathological is cut into slices:The formational situation and Pathological of optical microphotograph Microscopic observation calcium oxalate crystal change.
Wherein, renal tissue and oxalate crystal growth in healthy urine content use catalytic spectrophotometric, autogamy solvent, grope test bar
Feasible oxalic acid content assay method is stablized in part, foundation, and specific content of the test is as follows:
1.2.2.4.1 the measure of oxalate crystal growth in healthy urine and nephridial tissue medium-height grass acid content
Experiment uses the content of catalytic spectrophotometric oxalate crystal growth in healthy urine.In acid condition, oxalic acid can be catalyzed potassium chromate
Oxidation methyl is red, generates colourless product, and reaction equation is as follows:
Fading rate is represented by:
Using fixed time, under certain condition, the concentration of oxalic acid is directly proportional to the absorbance change of reaction system, this
For the basis of catalytic spectrophotometric oxalic acid.
Concrete operation method is as follows:
Each pipe operating method of table 1
Returned to zero instrument with distilled water, after 15min, each pipe adds 1.0mmol/L ZrOCl2·8H2O 1.0mL, shake up,
Terminating reaction.Determine absorbance, catalysing tube As, on-catalytic pipe is A0。
1.2.2.4.1.1 the determination of maximum absorption wavelength
Two 15mL tool plug test tubes are taken respectively as catalysing tube (S) and on-catalytic pipe (B), according to above-mentioned catalytic reaction method
Operation, length scanning is carried out in 400nm-700nm.As a result Fig. 1 is seen.As a result show, reaction system has maximum suction at 519nm
Receive, determine a length of 519nm of maximum absorption wave.
1.2.2.4.1.2 linear relationship is investigated
Precision weighs that oxalic acid is appropriate, and the stock solution that concentration is 1mmoL/L is made.It is appropriate that precision measures stock solution, dilute respectively
Release to the solution that concentration is 2,4,8,12,16,20 μm of oL/L, operated according to catalytic reaction method, absorbance is recorded, with concentration
(μm ol/L) is abscissa, lg (A0/ A) it is ordinate, try to achieve regression equation:Y=0.00390x+0.01054, r2=0.9993.
Result of the test shows, oxalic acid in the range of 0-20 μm of ol/L with lg (A0/ A) there is good linear relationship.Result of the test is shown in Table 2,
Fig. 2.
The oxalic acid standard curve of table 2
1.2.2.4.1.3 stability test
2 15mL tool plug test tubes are taken, as catalysing tube (S) and on-catalytic pipe (B), are operated according to catalytic reaction method, point
Not 0 after preparation, 10,20,30,40,50,60min determine the absorbance of each pipe, the RSD for measuring catalysing tube absorbance is
0.880%, the RSD of on-catalytic pipe absorbance is 0.619%, illustrates that reaction system is stable in 1 hour, the results are shown in Table 3.
The catalysing tube of table 3 is investigated with on-catalytic pipe color stability
1.2.2.4.1.4 precision test
10 μm of ol/L oxalic acid solution is taken, is operated according to catalytic reaction method, METHOD FOR CONTINUOUS DETERMINATION 6 times, records absorbance, is surveyed
The RSD for obtaining absorbance is 0.611% (n=6), meets the requirements, the results are shown in Table 4.
The precision test of table 4
1.2.2.4.1.5 recovery test
6 parts of urine sample for having predicted oxalic acid content is taken, precision measures 0.50mL, is separately added into sodium oxalate reference substance solution
0.50mL, rate of recovery need testing solution is made, with reference to catalytic reaction method, determination sample medium-height grass acid content, is calculated back respectively
Yield, as a result oxalic acid average recovery rate is that 101.3%, RSD is 0.858% (n=6), meets the requirements, the results are shown in Table 5.
The rate of recovery data of table 5
1.2.2.4.1.6 urine specimen and nephridial tissue medium-height grass acidity test
Urine pre-treating method:Urine sample 1.5mL is taken, adds activated carbon 0.25g, is decolourized, 10000r/min centrifugation 10min,
Filtration, 100 times of dilution are standby.Urine sample medium-height grass acid content is measured according to catalytic reaction operating method.
Renal tissue pre-treating method:Nephridial tissue is taken, it is accurately weighed, add hydrochloric acid:Ethanol:Water (34:120:46) solution
8mL, homogenate, 48h is stood, 3500r/min centrifuges 20min, and Aspirate supernatant is standby.With reference to oxalate crystal growth in healthy urine assay side
Method, determine nephridial tissue medium-height grass acid content.
1.2.2.4.2 SOD measure in nephridial tissue
Nephridial tissue is taken, it is accurately weighed, according to weight (g):Volume (mL)=1:9 ratio adds the physiology salt of 9 times of volumes
Water, shreds tissue, and ice-water bath prepares tissue homogenate, 3000r/min centrifugation 10min, takes supernatant.According to SOD kit sides
Method operates, and determines protein concentration, SOD inhibiting rates in supernatant, calculates SOD vigor.
1.2.2.4.3 the measure of calcium ion content in nephridial tissue
1.2.2.4.3.1 measuring principle
Calcium In Tissues ion combines in alkaline solution with methyl thymol blue (MTB), generates blue complex.Pass through
Colorimetric can calculate the content of calcium compared with the standard calcium equally handled.
1.2.2.4.3.2 operating method, as shown in table 6.
The calcium of table 6 determines operating method
Mix, after standing 5min, 610nm, 1cm optical paths, distilled water zeroing, determine each pipe absorbance.
Calculation formula:
1.2.2.4.3.3 the range of linearity
With reference to 2.2.2.4.3.2 operating methods, calcium ion canonical plotting is as shown in figure 3, standard calcium curve data such as table 7
It is shown, the calcium solution of various concentrations is separately added into, determines its absorbance.As a result show in the range of 0-5mmol/L, absorbance A
There is good linear relationship with calcium solution concentration.
The standard calcium curve data of table 7
1.3 statistical analysis technique
Experimental data is represented with mean ± standard deviation (± S), is handled using the statistical softwares of SPSS 22.0, and t- is used between group
Inspection is compared, P<0.05 expression difference has statistical significance, P<0.01 thinks with significant difference.
1.4 experimental result
1.4.1 ordinary circumstance observes result
Without animal dead in experimentation.Model control group Hair of Rats is not flourish, matt, and tired mind, activity subtracts
It is few.Shenshitong group and the basic, normal, high dosage group hair color of clear stone particle are fair and clear, glossy, mental status, and diet activity is normal.
After experiment, model control group body weight reduces (P compared with blank control group body weight conspicuousness<0.01), Shenshitong group, clear stone particle it is low,
Middle and high dosage group body weight increases in various degree compared with model control group, and difference has statistical significance (P<0.05).(it is shown in Table 8, figure
10)
Influence of the clear stone particle of table 8 to experimental rat body weight
Note:* the P compared with blank control group is represented<0.05, * * represents the P compared with blank control group<0.01, # expression and mould
Type control group compares P<0.05, ## represents the P compared with model control group<0.01.
1.4.2 blood biochemistry analysis result
Each group experimental rat blood biochemistry analysis result is shown in Table 9.Compared with blank control group, model control group urea BU, Cr,
Significantly rise (P<0.01);Compared with model control group, Shenshitong group rat blood serum urea substantially reduces (P<0.05), serum flesh
Acid anhydride declines unobvious.Clear basic, normal, high dosage group serum BU, Cr of stone particle significantly reduces (P compared with model control group<0.01).With
Blank control group is compared, model control group Ca2+Significantly rise (P<0.01), Shenshitong group, the clear basic, normal, high dosage group of stone particle
Ca2+(P is significantly reduced compared with model control group<0.01).(being shown in Table 9, Figure 11~Figure 13)
Influence of the clear stone particle preventing and treating administration of table 9 to calculus blood biochemistry of rats
Note:* the p compared with blank group is represented<0.05, * * represents the p compared with blank group<0.01, # expression and model control group
Compare p<0.05, ## represents the p compared with model group<0.01.
1.4.3 biochemical studies are urinated
Each group experimental rat urine biochemical studies are shown in Table 10.Compared with blank control group, model control group 24h urine volume,
Urinary calcium, urine oxalic acid significantly raise (P<0.05) pH value, is urinated, urine uric acid significantly reduces (P<0.01);Compared with model control group, kidney
The logical group rat 24h urine volume of stone, urine pH value dramatically increase (P<0.05) it is not notable, to urinate uric acid increase, urine oxalic acid, urinary calcium significantly reduce
(P<0.01).The basic, normal, high dosage group 24h urine volume of stone particle, urine pH value, urine uric acid dramatically increase compared with model control group clearly, urinate
Oxalic acid, urinary calcium reduce.(P<0.05).(being shown in Table 10, Figure 14~Figure 18)
The clear stone particle preventing and treating administration of table 10 is to the biochemical influence of calculus rat urine
Note:* the p compared with blank group is represented<0.05, * * represents the p compared with blank group<0.01, # expression and model control group
Compare p<0.05, ## represents the p compared with model group<0.01
1.4.4 calcium, oxalic acid, SOD measurement results in Kidney coefficients and nephridial tissue
As a result show:Model control group Kidney coefficients significantly raise (P compared with blank control group<0.01), Shenshitong group, clear stone
The basic, normal, high dosage group Kidney coefficients of particle have different degrees of reduction (P compared with model control group<0.05).Nephridial tissue super oxygen
Thing mutase measurement result represents that model control group SOD significantly reduces (P compared with blank control group<0.01), Shenshitong group, clear stone
The basic, normal, high dosage group SOD of particle has different degrees of rise (P compared with model control group<0.05).Model control group nephridial tissue
Interior calcium contains, oxalic acid content significantly rise (P compared with blank control group<0.01), calcium, oxalic acid contain in remaining each treatment group's nephridial tissue
Amount has different degrees of reduction.(being shown in Table 11, Figure 19~Figure 22)
Influence of the clear stone particle preventing and treating administration of table 11 to calculus renal tissues of rats
Note:* the p compared with blank group is represented<0.05, * * represents the p compared with blank group<0.01, # expression and model control group
Compare p<0.05, ## represents the p compared with model group<0.01.
1.4.5 Pathological inspection result
Blank control group experimental rat renal tissue structure is normal, epithelial cell marshalling, rule, has no expansion, becomes
Property oedema and calcium oxalate crystal distribution.Model control group rat kidney tissue cortex, medullary substance boundary are unintelligible;Epithelial cell is presented
Swelling, the denaturation of height;Lumen shape is irregular, and intracavitary is visible to be dispersed in or connect sheet of calcium oxalate crystals aggregation and sphacelus
Matter, visible a large amount of acute, the chronic inflammatory cell infiltrations of interstitial.It is epithelial cell swelling in Shenshitong group rat kidney tissue, bad
Extremely improve to a certain degree, calcium oxalate crystal distribution is reduced, and acute, chronic inflammatory cell infiltration occurs in part interstitial.Clear stone
It is accidental in Li Ge treatment groups rat kidney tissue to be dispersed in calcium oxalate crystal, cortex, medullary substance distinct;Epithelial cell is substantially without swollen
Swollen, tube chamber marshalling, interstitial is acute, chronic cell infiltration significantly mitigates compared with model control group.Clear stone particle is basic, normal, high
Obvious difference is not presented for dosage group.(see Fig. 4~Fig. 9)
1.5 conclusions are with discussing
Urolithiasis is urinary system common disease and frequently-occurring disease, wherein kidney stone incidence of disease highest, and 80% is calcinm oxalate calculus.
Clinically calcium in urine, oxalic acid, the phosphate of showing as are discharged the discharges such as increase, magnesium ion, citric acid, citric acid and reduced more, and
It is often accompanied by blood calcium rise.Kidney stone pathogenic factor and mechanism are more complicated, are that inherent cause and environmental factor etc. are coefficient
As a result.The pathogenesis and reason for meeting the animal model of mankind's kidney stone pathogenic process to illustrating kidney stone are established, searching has
The therapeutic scheme of effect is significant.Research is thought at present, and the supersaturation of calcium oxalate is the prerequisite that kidney stone is formed, and
By going through nucleation, growth, aggregation and a series of physical chemical process such as solid phase transformation.By increasing the intake of ethylene glycol,
Increase metabolism generation and the content of oxalate crystal growth in healthy urine of internal oxalic acid, promote the supersaturation of calcium oxalate.Ammonium chloride has kidney malicious
Property, can induced renal damage renal cells, cause its necrosis, come off, the formation for crystallization provides nucleus and is calcium oxalate
The adhesion of crystal provides condition.Meanwhile ammonium chloride can be acidified urine, promote the supersaturation of calcium oxalate.This experiment uses 1% chlorine
Change ammonium and combine 1% ethylene glycol duplication kidney calcinm oxalate calculus model.Test result indicates that model control group experimental rat serum calcium,
Creatinine, urea significantly increase, and oxalic acid, calcium content and pathological examination and blank group have significant difference in nephridial tissue.Represent big
The success of mouse kidney calcinm oxalate calculus model copy.
Traditional Chinese medicine thinks:Kidney stone belongs to the category of the illness such as " urolithiasis ", " sand leaching ", " blood strangury ", " pain in the back ".Clinical multilist
Now for the short puckery, sound of rain pattering shouting pain of urine frequency, dripping notably sediment outflow stone etc. is primary symptom not to the utmost, how touching the course of disease is repeatedly.Clear stone particle by
The taste traditional Chinese medicine extraction of pyrrosia lingua, Lygodium japonicum, oriental wormwood, the bark of official magnolia, rhizoma alismatis etc. 13 is refined to be formed, the work(of cooling blood and relieving pain treating stranguria with niter
Effect.In said preparation prescription, the analgesic of Lygodium japonicum inducing diuresis for treating strangurtia, pyrrosia lingua inducing diuresis for treating strangurtia, and energy cooling blood and hemostasis, Lygodium japonicum obtain pyrrosia lingua niter
And can stop blooding, for face lithiasis often with the disease of blood urine, the two niter is treating stranguria, cooling blood and relieving pain, is altogether monarch drug in a prescription.It is simultaneous that pivot stores inducing diuresis for treating strangurtia
There are heat-clearing, oriental wormwood clearing heat and promoting diuresis, rhizoma alismatis eliminating dampness and diuresis purges heat, and three makes damp and hot from urine to go out;Bark of official magnolia eliminating dampness promoting the circulation of qi, Fu
Siberian cocklebur inducing diuresis with bland drug and can invigoration spleen qi, so as to strengthen dampness removing function, fringed pink, imperata rhizome heat-clearing diuretic, cooling blood and hemostasis, help pyrrosia lingua with
Pivot storage, oriental wormwood, the work(of rhizoma alismatis, it is altogether ministerial drug.The shell of areca nut, Radix Astragali promoting the circulation of qi inducing diuresis to remove edema, root of three-nerved spicebush promoting qi circulation and relieving pain are adjutant, San Zhehang
Gas is helped below stone, is arranged stone and is gone out.Radix glycyrrhizae coordinating the drug actions of a prescription, relieving spasm to stop pain, and energy heat-clearing, are adjuvant.
Test result indicates that calcium, oxalic acid content significantly reduce in clear each treatment group's nephridial tissue of stone particle, pathological examination result
It has been shown that, in kidney calcium oxalate crystal distribution reduce, tubular ectasia, come off, necrosis is obviously improved.Clear stone particle is prompted effectively to press down
The formation and promotion calculus discharge of calcinm oxalate calculus processed.Meanwhile urinate Analysis of Biochemical result and show that clear stone particle can be carried significantly
High 24h urine volume, promote urine uric acid excretion, alkalized urine.Increase urine volume and alkalized urine can reduce calcium oxalate supersaturation and separate out, and add
By force to the souring of urinary tract, calcium oxalate crystals are reduced to the adhesive attraction of renal cells, suppress the formation of calculus.Urine
Acid can be combined with calcium oxalate crystals, promote the growth of calcium oxalate crystals.Clear stone particle urinates the excretion of uric acid by increasing, and reduces urine
Acid reduces the formation of calcium oxalate crystal to the facilitation of calculus.
Clear stone particle can significantly reduce serum calcium ion, creatinine, urea content, alleviate impaired renal function symptom, improve kidney
Bead filtering function, protect renal function.The oxalic acid of high concentration can cause renal cells to produce reactive oxygen species, cause thin
Born of the same parents' apoptosis, necrosis and cell membrane surface characteristic changing, promote calculus.Clear stone particle can improve polyphenoils in nephridial tissue
SOD level, promote oxalic acid excretion, the damage of renal cells is reduced, so as to suppress the formation of calcium oxalate crystals.
In summary, clear stone particle can effectively reduce calcium in nephridial tissue, oxalic acid content, protect renal cells, promote
Enter the excretion of uric acid, alkalized urine, increase urine discharge capacity, so as to suppress the formation of calcinm oxalate calculus.Its mechanism of action may with it is clear
Stone particle can increase mortifier level in urine, reduce calculus promoting factor, increase renal cells anti-oxidation stress
It is horizontal relevant.
Embodiment 2
A kind of clear stone particle, include the component of following parts by weight:320 parts of Lygodium japonicum, 240 parts of pyrrosia lingua, 240 parts of the bark of official magnolia,
240 parts of the Fen , pivots of Poria cocos 320 storage, 240 parts of oriental wormwood, 320 parts of rhizoma alismatis, 240 parts of fringed pink, 240 parts of rhizoma imperatae, 320 parts of the shell of areca nut are yellow
160 parts of 240 parts of stilbene, 320 parts of the root of three-nerved spicebush and honey-fried licorice root.
A kind of preparation method of clear stone Chinese medicine composition, comprises the following steps:
Step 1: in terms of parts by weight, the bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis are taken, it is small to add 10 times of amount 70% alcohol reflux extractions 2
When, filtration, filtrate recycling ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25.
Step 2: in terms of parts by weight, the dregs of a decoction and Lygodium japonicum, pyrrosia lingua, Poria cocos, pivots storage, the mattress that are filtrated to get in step 1
Old, fringed pink, rhizoma imperatae, the shell of areca nut, the Radix Astragali and honey-fried licorice root mixing, 10 times of amount water are added to soak 2 hours, decoction is secondary, and 2 is small every time
When, decocting liquid filtration, merge with the filtrate in step 1, and concentrate, let cool afterwards, then plus 80% ethanol reaches alcohol content
60%, stand, filtration, filtrate recycling ethanol is simultaneously concentrated into the leaching that relative density at temperature 50 C~55 DEG C is 1.20~1.25
Cream, merge mixing with the above-mentioned bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis medicinal extract.
Step 3: adding dextrin and flavouring into above-mentioned composition, mix, pelletize, dry, sugar-free particle 1000 is made
Part, obtain clear stone particle.
Embodiment 3
A kind of clear stone particle, include the component of following parts by weight:480 parts of Lygodium japonicum, 360 parts of pyrrosia lingua, 360 parts of the bark of official magnolia,
360 parts of the Fen , pivots of Poria cocos 480 storage, 360 parts of oriental wormwood, 480 parts of rhizoma alismatis, 360 parts of fringed pink, 360 parts of rhizoma imperatae, 480 parts of the shell of areca nut are yellow
240 parts of 360 parts of stilbene, 480 parts of the root of three-nerved spicebush and honey-fried licorice root.
A kind of preparation method of clear stone Chinese medicine composition, comprises the following steps:
Step 1: in terms of parts by weight, the bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis are taken, 8 times of 40% alcohol refluxs of amount is added and extracts 2 hours,
Filtration, filtrate recycling ethanol are simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25.
Step 2: in terms of parts by weight, the dregs of a decoction and Lygodium japonicum, pyrrosia lingua, Poria cocos, pivots storage, the mattress that are filtrated to get in step 1
Old, fringed pink, rhizoma imperatae, the shell of areca nut, the Radix Astragali and honey-fried licorice root mixing, 8 times of amount water are added to soak 2 hours, decoction is secondary, 2 hours every time,
Decocting liquid filters, and merges with the filtrate in step 1, and concentrates, and lets cool afterwards, and then plus 50% ethanol makes alcohol content up to 60%, quiet
Put, filter, filtrate recycling ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25, and above-mentioned
The bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis medicinal extract, which merge, to be mixed.
Step 3: adding dextrin, sugar and flavouring into above-mentioned composition, mix, pelletize, dry, granules of sugar is made
1000 parts, obtain clear stone particle.
Embodiment 4
A kind of clear stone particle, include the component of following parts by weight:
200 parts of Lygodium japonicum, 150 parts of pyrrosia lingua, 150 parts of the bark of official magnolia, 150 parts of the Fen , pivots of Poria cocos 200 storage, 150 parts of oriental wormwood, rhizoma alismatis 200
100 parts of part, 150 parts of fringed pink, 150 parts of rhizoma imperatae, 200 parts of the shell of areca nut, 150 parts of the Radix Astragali, 200 parts of the root of three-nerved spicebush and honey-fried licorice root.
A kind of preparation method of clear stone Chinese medicine composition, comprises the following steps:
Step 1: in terms of parts by weight, the bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis are taken, 5 times of 80% alcohol refluxs of amount is added and extracts 2 hours,
Filtration, filtrate recycling ethanol are simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25.
Step 2: in terms of parts by weight, the dregs of a decoction and Lygodium japonicum, pyrrosia lingua, Poria cocos, pivots storage, the mattress that are filtrated to get in step 1
Old, fringed pink, rhizoma imperatae, the shell of areca nut, the Radix Astragali and honey-fried licorice root mixing, 5 times of amount water are added to soak 2 hours, decoction is secondary, 2 hours every time,
Decocting liquid filters, and merges with the filtrate in step 1, and concentrates, and lets cool afterwards, and then plus 95% ethanol makes alcohol content up to 60%, quiet
Put, filter, filtrate recycling ethanol is simultaneously concentrated into the medicinal extract that relative density at temperature 50 C~55 DEG C is 1.20~1.25, and above-mentioned
The bark of official magnolia, the root of three-nerved spicebush and rhizoma alismatis medicinal extract, which merge, to be mixed.
Step 3: adding dextrin and flavouring into above-mentioned composition, mix, pelletize, dry, sugar-free particle 1000 is made
Part, obtain clear stone particle.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details and shown here as the legend with description.