CN103054929A - Rhodiola extract and pharmaceutical composition thereof as well as application of rhodiola extract in treatment of fatty liver disease - Google Patents
Rhodiola extract and pharmaceutical composition thereof as well as application of rhodiola extract in treatment of fatty liver disease Download PDFInfo
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Abstract
The invention discloses a rhodiola extract and a pharmaceutical composition thereof and application of the rhodiola extract in treatment of a fatty liver disease. A preparation method for an effective part of rhodiola is characterized by comprising the following steps of: (A) crushing dry rhizomes of a rhodiola plant, performing extraction by using an organic solvent, and recovering the solvent from the extraction liquid under reduced pressure to obtain an extract; adjusting the extract to a proper density, performing water deposition in pure water, filtering the sediment, and concentrating the filtrate under reduced pressure to obtain a total rhodiola extract; and (B) separating the total rhodiola extract by using macromolecular adsorption resin, performing gradient elution by using a water or ethanol system, concentrating the eluent under reduced pressure, performing freeze drying, thus obtaining the effective part of the rhodiola.
Description
Technical field
The present invention relates to a kind of Radix Rhodiolae extract, the purposes of its pharmaceutical composition in preparation treatment fatty liver medicine and health product that obtains of from Rhodida plant, separating, belong to field of medicaments.
Background technology
Rhodida plant is Crassulaceae (Crassulaceae) rhodiola (Rhodiola) plant, and medicinal history is long, hides name " Suo Luomabao "." Jingzhubencao " record, the effect of Radix Rhodiolae are to support lung, heat clearing away, nourishing vigour etc.Tibet Radix Rhodiolae commonly used among the people is treated the diseases such as hemoptysis, spitting of blood, pneumonia cough and women leucorrhea.The Shennong's Herbal record, the large showing tremendous enthusiasm skin ulcer of Radix Rhodiolae master, the evil gas of body dysphoria with smothery sensation, all insect poison crust skin ulcers, the cold and heat migratory arthralgia, the various symptoms and signs of deficiency flower, hostess's whitish metrorrhagia, making light of one's life by commiting suicide makes eye bright, and the logical god of clothes is not old for a long time.Claim in the Compendium of Material Medica that Radix Rhodiolae is " book on Chinese herbal medicine is top grade ", as the strengthening by means of tonics medicine, allaying tiredness, the cold resisting.Commonly used its among the people fried in shallow oil water or steeps in wine to eliminate the winter tired and the opposing high and cold mountain area that heavy physical labour brings cold.The kind of contained chemical composition is more in the Rhodida plant, and separating the chemical compound that obtains according to bibliographical information from Rhodida plant approximately has more than 100, can roughly be divided into eight compounds, it is terpenoid, flavonoid, cyanogen glycoside, benzyl ethylene, phenethyl alcohol glycoside and benzylalcohol glycoside, phenolic acids, lignanoids and other class, wherein phenolic acids, the phenethyl alcohol glycoside compounds is the total composition in the Rhodida plant, and content is higher in plant.
Fatty liver is the pathological state of fat abnormal accumulation in hepatocyte of causing of a kind of multi-pathogenesis.Total fat mass in normal person's the liver, account for liver heavy 5%, include phospholipid, triglyceride, fat acid, cholesterol and cholesterol ester.According to intrahepatic fat content be divided into slightly, moderate, severe fatty liver.Severe fatty liver mainly is that triglyceride and fat acid content increase.Diagnosis of Fatty liver is learned or liver biopsy pathological examination according to liver imagings such as B ultrasonic.Change process according to liver histopathology, fatty liver can be divided into simple fatty liver, fat hepatitis and fatty cirrhosis.
Up to the present; Western medicine there is no lipotropic active drug; be main mainly with symptomatic treatment; such as the protection such as vitamin B, C, E, lecithin, ursodesoxycholic acid, silymarin, inosine, coenzyme A, reduced glutathion, taurine, carnitine Orotate hepatocyte, antioxidant, and the fat-reducing medicament such as bifendate.The medicine of Chinese herb prevention fatty liver is main mainly with long-time conditioning, as contains Gan Beier of the Chinese medicine ingredients such as Rhizoma Curcumae Longae etc., the blood fat reducing medicines such as Radix Polygoni Multiflori Preparata and Fructus Crataegi etc.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of Radix Rhodiolae effective site with effect for the treatment of fatty liver.
The technical problem to be solved in the present invention provides a kind of pharmaceutical composition, and it comprises described Radix Rhodiolae effective site and pharmaceutically acceptable carrier.
The technical problem to be solved in the present invention provides described Radix Rhodiolae effective site and treats and/or prevents application in the medicine of fatty liver in preparation.
The technical problem to be solved in the present invention provides the application of described Radix Rhodiolae effective site in preparation fatty liver health product.
The technical problem to be solved in the present invention provides described Radix Rhodiolae effective site and treats and/or prevents application in the compound recipe of fatty liver in preparation.
For solving technical problem of the present invention, the present invention adopts following technical scheme:
A kind of preparation method of Radix Rhodiolae effective site is characterized in that may further comprise the steps:
(1) dry rhizome of Rhodida plant is pulverized, extracted with methanol, ethanol or acetone, the extracting solution reclaim under reduced pressure is held agent, gets extractum; Extractum is adjusted to suitable density, carry out water precipitating in pure water, with sedimentation and filtration, filtrate decompression is concentrated, gets the Radix Rhodiolae total extract;
(2) the Radix Rhodiolae total extract is separated with macroporous adsorbent resin, water, ethanol system carry out gradient elution, with the eluent concentrating under reduced pressure, get Radix Rhodiolae effective site after the lyophilization.
Wherein, extraction solvent concentration is preferably 30% to 95% in the step (1), and its consumption is preferably 6 to 12 times of medical material weight; Leaching process can repeat 2 to 3 times, and each extraction time is 1 to 3 hour; Preferably extractum density is adjusted into 1.15 to 1.19; The water precipitating step left standstill under 4 to 10 ℃ 24 to 96 hours, and the amount of required pure water is 4 to 10 times of extractum weight;
Macroporous adsorbent resin described in the step (2) is low pole or Microporous resin, and its model is selected from Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, DM301, NKA-9, AB-8, D101, DM130, LSA-10, LSA-20, DM-18 and D312 etc.; Be preferably Diaion HP-20, DiaionHP-40, D101, AB-5; Gradient elution at first water carries out eluting, then carries out gradient elution with ethanol, water mixed solvent; Preferably adopt successively water, 5% ethanol, 50% ethanol and four gradients of 95% ethanol to carry out eluting; The eluant consumption of each gradient is preferably 2 to 4 column volumes; The eluate that preferred collection 50% ethanol elution obtains.
Rhodida plant of the present invention includes but not limited to Radix Rhodiolae, Rhodiola kirilowii (Regel) Maxim., Radix Rhodiolae, rose-red Herba hylotelephii erythrosticti, Rhodiola atuntsuensis, Rhodiola himalensis, Rhodiola phariensis, Rhodiola henryi, Rhodiola quadrifida, Rhodiola yunnanensis, Rhodiola wallichiana var. cholaensis, Rhodiola fastigiata, Rhodiola Crassulaceae, Radix Rhodiolae, Rhodiola dumulosa and Rhodiola sachalinensis.
The present invention relates to the Radix Rhodiolae effective site that obtained by said method.
The present invention relates to a kind of pharmaceutical composition, comprise described Radix Rhodiolae effective site and pharmaceutically acceptable carrier.
The invention still further relates to and contain as the Radix Rhodiolae effective site of active ingredient and the pharmaceutical composition of conventional medicine excipient or adjuvant.Usually Radix Rhodiolae effective site accounts for 0.1～95% of pharmaceutical composition gross weight.
The present invention also provides a kind of pharmaceutical composition, and it comprises Radix Rhodiolae effective site and the pharmaceutically acceptable carrier as active component of medicine effective dose.
Pharmaceutical composition of the present invention can be according to method preparation well known in the art.When being used for this purpose, if necessary, Radix Rhodiolae effective site of the present invention and one or more solids or liquid medicine excipient and/or adjuvant can be combined, make the suitable administration form or the dosage form that can be used as people's medicine or veterinary drug use.
Radix Rhodiolae effective site of the present invention or contain its pharmaceutical composition can the unit dosage form administration, route of administration can be intestinal or non-intestinal, such as oral, muscle, subcutaneous, nasal cavity, oral mucosa, eye, lung, skin, vagina, peritoneum, rectum etc., preferred oral administration.
Radix Rhodiolae effective site of the present invention or the route of administration that contains its pharmaceutical composition can be drug administration by injection.Injection comprises intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, lumbar injection and acupoint injection therapy etc.
Form of administration can be liquid dosage form, solid dosage forms or semisolid dosage form.Liquid dosage form can be solution (comprising true solution and colloid solution), Emulsion (comprising oil-in-water type, water-in-oil type and emulsion), suspensoid, injection (comprising aqueous injection, injectable powder and transfusion), eye drop, nasal drop, lotion and liniment etc.Solid dosage forms can be tablet (comprising ordinary tablet, enteric coatel tablets, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, oral cavity disintegration tablet), capsule (comprising hard capsule, soft capsule, enteric coated capsule), granule, powder, micropill, drop pill, suppository, membrane, paster, the agent of gas (powder) mist, spray etc.; Semisolid dosage form can be ointment, gel, paste etc.
Radix Rhodiolae effective site of the present invention can be made ordinary preparation, also can be slow releasing preparation, controlled release preparation, targeting preparation and various particulate delivery system.
For the unit form of administration is made tablet, can be widely used various excipient well known in the art, comprise diluent, adhesive, wetting agent, disintegrating agent, lubricant, fluidizer.Diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline Cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate etc.; Wetting agent can be water, ethanol, isopropyl alcohol etc.; Binding agent can be that starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline Cellulose, mucialga of arabic gummy, gelatine size, sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl emthylcellulose, ethyl cellulose, acrylic resin, carbomer, polyethylene adjoin pyrrolidone, poly-second two propanol etc.; Disintegrating agent can be that dried starch, microcrystalline Cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyethylene adjoin pyrrolidone, cross-linking sodium carboxymethyl cellulose, carboxymethyl starch sodium, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, dodecyl sodium sulfate; Lubricant and fluidizer can be Pulvis Talci, silicon dioxide, stearate, tartaric acid, liquid paraffin, Polyethylene Glycol etc.
Tablet further can also be made coated tablet, for example sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablet and multilayer tablet.
For pill is made in the administration unit, can be widely used various carrier well known in the art.Example about carrier is, for example diluent and absorbent are such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire 44/14, Kaolin, Pulvis Talci etc.; Binding agent is such as arabic gum, yellow Bodhisattva's glue, gelatin, ethanol, Mel, liquid sugar, rice paste or batter etc.; Disintegrating agent is such as agar powder, dry starch, alginate, dodecyl sodium sulfate, methylcellulose, ethyl cellulose etc.
For suppository is made in the administration unit, can be widely used various carrier well known in the art.Example about carrier is, such as the ester of Polyethylene Glycol, lecithin, cocoa butter, higher alcohol, higher alcohol, gelatin, semi-synthetic glyceride etc.
For capsule is made in the administration unit, Radix Rhodiolae effective site of the present invention is mixed with above-mentioned various carriers, and the mixture that will obtain thus places hard gelatine capsule or soft capsule.Also effective ingredient can be made microcapsule, be suspended in and form suspensoid in the aqueous medium, in the hard capsule of also can packing into or make injection and use.
For example, Radix Rhodiolae effective site of the present invention is made injection preparation, such as solution, suspensoid solution, Emulsion, lyophilized injectable powder, this preparation can be moisture or non-water, can contain acceptable carrier, diluent, binding agent, lubricant, antiseptic, surfactant or dispersant on a kind of and/or multiple pharmacodynamics.Can be selected from water, ethanol, Polyethylene Glycol, l such as diluent, the isooctadecanol of ammediol, ethoxylation, the isooctadecanol of polyoxy, Polyoxyethylene Sorbitol Fatty Acid Esters etc.In addition, to ooze injection in order preparing etc., can in injection preparation, to add an amount of sodium chloride, glucose or glycerol, in addition, can also add conventional cosolvent, buffer agent, pH adjusting agent etc.These adjuvants are that this area is commonly used.
In addition, such as needs, also can in pharmaceutical preparation, add coloring agent, antiseptic, spice, correctives, sweeting agent or other material.
For reaching the medication purpose, strengthen therapeutic effect, Radix Rhodiolae effective site of the present invention or pharmaceutical composition can be with any known medication administrations.
The dosage of pharmaceutical composition of the present invention depends on many factors, for example to prevent or treat character and the order of severity of disease, the sex of patient or animal, age, body weight, personality and individual reaction, route of administration, administration number of times, therapeutic purposes, therefore therapeutic dose of the present invention can have large-scale variation.In general, the using dosage of Chinese materia medica composition of the present invention is well known to a person skilled in the art.Can be according to the present invention contained actual drug quantity in the last preparation in the pharmaceutical composition, in addition suitable adjustment to reach the requirement of its treatment effective dose, is finished prevention of the present invention or therapeutic purposes.The suitable dose scope of the every day of Radix Rhodiolae effective site of the present invention: the consumption of Radix Rhodiolae effective site of the present invention is 1～1000mg/Kg body weight, above-mentioned dosage can the single dose form or is divided into several, for example two, three or four dosage form administrations, this depends on administration doctor's clinical experience and comprises the dosage regimen of using other treatment means.Each treats that required accumulated dose can be divided into repeatedly or by the dose administration.Chemical compound of the present invention or pharmaceutical composition can be taken separately, or merge use and adjust dosage with other treatment medicine or symptomatic drugs.
Useful technique effect
1, Radix Rhodiolae extract of the present invention has the effect of obvious treatment fatty liver, comprising reducing liver fat content, improves fatty liver pathology and changes, improves the Liver Microcirculation effect.
2, by macroporous resin Radix Rhodiolae extract is carried out enrichment, a large amount of carbohydrate contents and some other impurity component have been removed, greatly reduce the dosage of use, the effective site of preparation gained is the 1%-3% of crude drug in whole only, is conducive to the Chinese patent medicine that further exploitation is prepared into various dosage forms.
3. Radix Rhodiolae extract provided by the present invention has the advantages such as preparation technology is simple, easy to operate, production cost is low, safe and reliable, and whole process meets the requirement of environmental protection.
Description of drawings
Fig. 1. Radix Rhodiolae extract is on the impact of mice fatty liver content of triglyceride
N=8.
#P＜0.05vs Normal group;
* *P＜0.001vs model control group.
Fig. 2. Radix Rhodiolae extract is on the adipohepatic impact of KKAy mice (HE dyeing; * 40)
Fig. 3. Radix Rhodiolae extract is measured the impact of mice fatty liver and liver central vein caliber
A. liver surface fat drips and central vein; B. the measurement of liver central vein caliber and statistical analysis;
##, p＜0.01vs Normal group;
*, p＜0.01vs model control group.
Fig. 4. Radix Rhodiolae extract is on the impact of the open number of Models of Fatty Liver mouse liver sinus hepaticus
###, p＜0.001vs Normal group;
*, p＜0.01vs model control group.
Fig. 5. Radix Rhodiolae extract is on the impact of fatty liver mouse liver central vein, sinus hepaticus leukocyte rolling
A. the leukocyte rolling of the bright 6G labelling of Luo Da in the central vein; B. the bright 6G labelling of Luo Da white thin in the sinus hepaticus
The born of the same parents .### that rolls, p＜0.001vs Normal group;
* *, p＜0.001vs model control group.
Fig. 6. Radix Rhodiolae extract is on the impact of model mice liver central vein red cell velocity, shearing
A, the central vein red cell velocity; B, central vein erythrocyte shearing.###, p＜0.001vs is normal
Matched group;
*,
* *, p＜0.05,0.001vs model control group.
The specific embodiment
The experiment of the following examples and pharmacologically active is used for further specifying the present invention, but this and do not mean that any limitation of the invention
The preparation of Radix Rhodiolae effective site
Embodiment 1
Adopt dry Radix Rhodiolae rhizome as crude drug in whole, after pulverizing with 80% alcohol reflux 3 times, each consumption is 8 times of crude drug in whole weight, extraction time is each 2 hours, extracting solution is merged rear the filtration, and filtrate decompression concentration and recovery ethanol gets extractum to distinguishing the flavor of without alcohol, regulating extractum density is 1.16, the water that adds 5 times of amounts carries out water precipitating, leaves standstill 72 hours under 5 ℃, with sedimentation and filtration, carrying out the HP-20 macroporous adsorbent resin after filtrate decompression is concentrated separates, adopt pure water, 50% ethanol, three gradients of 95% ethanol are carried out eluting, with the eluent concentrating under reduced pressure of 50% ethanol, after lyophilization, obtain the effective site of Radix Rhodiolae.
Adopt dry Rhodiola kirilowii (Regel) Maxim. rhizome as crude drug in whole, after pulverizing with 95% alcohol reflux 2 times, each consumption is 7 times of crude drug in whole weight, extraction time is each 3 hours, filters after extracting solution is merged, and filtrate decompression concentration and recovery ethanol is to nothing alcohol flavor, get extractum, regulating extractum density is 1.18, and the water that adds 8 times of amounts carries out water precipitating, leaves standstill 96 hours under 5 ℃, with sedimentation and filtration, carry out the HP-40 macroporous adsorbent resin after filtrate decompression is concentrated and separate, adopt pure water, 5% ethanol, 50% ethanol, four gradients of 95% ethanol are carried out eluting, with the eluent concentrating under reduced pressure of 50% ethanol, obtain the effective site of Rhodiola kirilowii (Regel) Maxim. after lyophilization.
Embodiment 3
Adopt dry Radix Rhodiolae rhizome as crude drug in whole, extract 2 times with 70% methanol eddy after pulverizing, each consumption is 8 times of crude drug in whole weight, extraction time is each 2 hours, filters filtrate decompression concentration and recovery methanol after extracting solution is merged, get extractum, regulating extractum density is 1.17, and the water that adds 6 times of amounts carries out water precipitating, leaves standstill 48 hours under 5 ℃, with sedimentation and filtration, carry out the D101 macroporous adsorbent resin after filtrate decompression is concentrated and separate, adopt pure water, 5% ethanol, 50% ethanol, four gradients of 95% ethanol are carried out eluting, and the eluent concentrating under reduced pressure of 50% ethanol obtains the effective site of Radix Rhodiolae after lyophilization.
Embodiment 4
Adopt dry Radix Rhodiolae rhizome as crude drug in whole, after pulverizing with acetone-water (7: 3) reflux, extract, 3 times, each consumption is 10 times of crude drug in whole weight, extraction time is each 2 hours, filters filtrate decompression concentration and recovery acetone after extracting solution is merged, get extractum, regulating extractum density is 1.18, and the water that adds 8 times of amounts carries out water precipitating, leaves standstill 72 hours under 5 ℃, with sedimentation and filtration, carry out the AB-8 macroporous adsorbent resin after filtrate decompression is concentrated and separate, adopt pure water, 5% ethanol, 50% ethanol, four gradients of 95% ethanol are carried out eluting, with the eluent concentrating under reduced pressure of 50% ethanol, obtain the effective site of Radix Rhodiolae after lyophilization.
The pharmacologically active experiment
Test example 1. Rhodiola crenulata extracts reduce the effect of fatty liver lipid content
Experimental technique: use high-sugar-fat-diet, feed C57BL mouse and form the fatty liver mouse model.Animal pattern is divided into 4 groups at random, be respectively model contrast, fenofibrate and Radix Rhodiolae extract-L, Radix Rhodiolae extract-H group, respectively oral solvent 0.5%CMC-Na, positive control drug fenofibrate 100mg/kg and Rhodiola crenulata extract 200,400mg/kg.Simultaneously, establish with crowd intact animal as Normal group.2 weeks of successive administration, behind the animal overnight fasting, sacrificed by decapitation, the sharp separation liver is made homogenate and is measured content of triglyceride.
Experimental result: compare with Normal group, model control group mouse liver content of triglyceride significance raises.Compare with model control group, Radix Rhodiolae extract-L, Radix Rhodiolae extract-H group liver tg content have reduced respectively 68.0%, 67.5%, have illustrated that FF3 has the effect (seeing Fig. 1) of obvious reduction obesity mice liver TG content.
Test example 2 Rhodiola crenulata extracts improve fatty liver pathology variation effect
Experimental technique: 3 groups of type 2 diabetes mellitus KKAy mices are respectively model contrast, fenofibrate and Radix Rhodiolae extract group, respectively oral solvent 0.5%CMC-Na, positive control drug fenofibrate 100mg/kg and Rhodiola crenulata extract 200mg/kg.Simultaneously, establish normal C57BL mouse as Normal group.Successive administration is after 2 weeks, sacrificed by decapitation, and the sharp separation liver, paraformaldehyde is liquid-solid fixed, carries out pathological examination.Pathological section is carried out the sxemiquantitative marking of liver lipid accumulation, and carry out statistical analysis.
Experimental result:
The Normal group mouse liver is no abnormal; The pathological characters of typical fatty liver has appearred in the liver of KKAy model control group mice, and main manifestations is for big or small mixed type steatosis occurring around central vein; Fenofibrate has been brought into play obvious improvement effect for steatosis.Radix Rhodiolae extract becomes for fat has also brought into play obvious improvement effect, and fat vacuole obviously reduces (seeing Fig. 2).
Model control group mouse liver content of triglyceride significance raises.Compare with model control group, Radix Rhodiolae extract-L, Radix Rhodiolae extract-H group liver tg content have reduced respectively 68.0%, 67.5%, have illustrated that FF3 has the effect of obvious reduction obesity mice liver TG content.Compare with group,
Obtain the hepatic steatosis scoring according to mice hepatic steatosis sxemiquantitative standards of grading, the result shows (table 1.), and the hepatic steatosis scoring of model control group is apparently higher than Normal group.Compare with model control group, the hepatic steatosis scoring of fenofibrate group mice has reduced by 17.9%; The scoring of Radix Rhodiolae extract group mice hepatic steatosis has reduced by 20.5%.
Table 1. Radix Rhodiolae extract is on adipohepatic impact
N=8.
###P＜0.001vs Normal group;
*P＜0.05,
*P＜0.01vs model control group.
Test example 3 Rhodiola crenulata extracts improve the effect of fatty liver microcirculation disturbance
Experimental technique:
Male 4 age in week the C57BL/6J mice, induce through high fat diet to form the non-alcoholic fatty liver disease model.Model mice is divided into 3 groups by blood lipid level: model control group, fenofibrate group, Radix Rhodiolae extract group give respectively oral 0.5% sodium carboxymethyl cellulose, positive control drug fenofibrate 100mg/kg, Radix Rhodiolae extract 200mg/kg.Establish simultaneously C57BL/6J mice to normal forage feed as Normal group.
3 weeks of successive administration.After the anesthesia of 20% urethane, expose liver surface, carry out observing at the liver microcirculation of body, under 10 times of mirrors, with the blood circumstance of colored CCD record liver edge central vein, more than the video recording 10s, be used for the subsequent result analysis.Every animal is chosen 3 different central veins at random, and with Imagepro-plus6.0 software analysis central vein caliber, every central vein is surveyed 3 times, gets average.And excite at mercury lamp under the condition of (455-492nm), record the sinus hepaticus number that directly links to each other with central vein under 20 times of mirrors.Every animal is chosen 3 visuals field at random, gets average.
With the bright 6G labelling of Luo Da leukocyte, leukocyte with the leukocyte of the leukocyte rolling of 10 seconds by the same cross section of central vein, represents with the leukocyte count (individual) that rolls/200m central vein in the video recording of playback; Take the leukocyte that 10 seconds, the leukocyte by sinus hepaticus in 200m * 300m visual field rolled in the sinus hepaticus, represent with the leukocyte count (individual) that rolls/visual field, every animal is chosen 3 visuals field at random, gets average.
With high-speed camera (FASTCAM-ultima APX, photron, Japan), to get speed, the red cell velocity of records center vein 2000 width of cloth/seconds.During measurement, the image of blood flow is measured red cell velocity at the speed playback of 25 width of cloth/seconds with Image-Pro Plus 6.0 softwares.Represent mice central vein blood flow rate with mm/s, choose 3 different central veins, survey 3 times, go average for every.Utilize the caliber of red cell velocity and central vein to calculate the erythrocyte shearing.The computing formula of erythrocyte shearing :=8 * (flow velocity/caliber) (S-1).
Experimental result:
Compare with Normal group, the visible a large amount of fat in model control group mouse liver surface drips the sample projection, the obvious constriction of the caliber of central vein (seeing Fig. 3 A); With model control group relatively, fenofibrate group, the visible a large amount of fat of Radix Rhodiolae extract group liver surface drip the sample projection be improved significantly, the caliber of central vein increases to some extent; Warp is measured and statistical analysis the central vein caliber, result's demonstration, and model control group mice central vein caliber has on average reduced by 13.0% than Normal group; After fenofibrate, Radix Rhodiolae extract treatment, compare with model control group, the central vein caliber of fenofibrate group, Radix Rhodiolae extract group mice has increased respectively by 13.7%, 15.0% (seeing Fig. 3 B).
Excite under the condition of (455-492nm) at mercury lamp, record the sinus hepaticus number that directly links to each other with central vein under 20 times of mirrors.The result shows.Compare with Normal group, the average sinus hepaticus of model control group mice has been opened decreased number 2.8 times.Compare with model control group, the open number of the average sinus hepaticus of fenofibrate group and Radix Rhodiolae extract group has increased respectively by 69.4% and 64.5% (Fig. 4)
Compare with Normal group, the leukocyte number of the bright 6G labelling of the Luo Da that the model control group animal rolls in central vein and sinus hepaticus has increased respectively 9.3 times and 5.9 times.After treatment, compare with model control group, the leukocyte number that rolls in central vein of fenofibrate group and Radix Rhodiolae extract group mice has reduced respectively by 83.5% and 66.6% (Fig. 5 A); The leukocyte number that rolls in sinus hepaticus has reduced respectively by 84.0% and 69.7% (Fig. 5 B).
Red cell velocity with high-speed camera records center vein.The result shows (Fig. 6 A, B), compares with Normal group, and the red cell velocity of model control group mice and erythrocyte shearing have reduced respectively 3.1 times and 2.6 times.With model control group relatively, the red cell velocity of Radix Rhodiolae extract group has accelerated 53.2%, the erythrocyte shearing has increased by 32.9%; The red cell velocity of fenofibrate group has accelerated 35.8%, and the erythrocyte shearing is then without significant change.
The above results demonstration, Radix Rhodiolae extract has the remarkable mice non-alcoholic fatty liver disease microcirculation disturbance effect that high fat diet causes that improves.
Claims (10)
1. the preparation method of a Radix Rhodiolae effective site is characterized in that may further comprise the steps:
(A) dry rhizome of Rhodida plant is pulverized, extracted with organic solvent, the extracting solution decompression and solvent recovery gets extractum; Extractum is adjusted to suitable density, carry out water precipitating in pure water, with sedimentation and filtration, filtrate decompression is concentrated, gets the Radix Rhodiolae total extract;
(B) the Radix Rhodiolae total extract is separated with macroporous adsorbent resin, water, ethanol system carry out gradient elution, with the eluent concentrating under reduced pressure, get Radix Rhodiolae effective site after the lyophilization.
2. according to claim 1 preparation method is characterized in that, the organic solvent described in the steps A is selected from methanol, ethanol or acetone.
3. according to claim 1 preparation method is characterized in that, extraction solvent concentration is that volume ratio 30% is to 95% organic solvent/water volume ratio in the steps A; Its consumption is 6 to 12 times of medical material weight; Leaching process can repeat 2 to 3 times, and each extraction time is 1 to 3 hour; Preferably extractum density is adjusted into 1.15 to 1.19; The water precipitating step left standstill under 4 to 10 ℃ 24 to 96 hours, and the amount of required pure water is 4 to 10 times of extractum weight.
4. according to claim 1 preparation method is characterized in that, the macroporous adsorbent resin described in the step B is low pole or Microporous resin.
5. according to claim 1 preparation method, it is characterized in that, the model of the macroporous adsorbent resin described in the step B is selected from Diaion HP-10, Diaion HP-20, Diaion HP-30, Diaion HP-40, Diaion HP-50, DM301, NKA-9, AB-8, D101, DM130, LSA-10, LSA-20, DM-18 and D312.
6. according to claim 1 preparation method, it is characterized in that, the Rhodida plant described in the steps A is selected from Radix Rhodiolae, Rhodiola kirilowii (Regel) Maxim., Radix Rhodiolae, rose-red Herba hylotelephii erythrosticti, Rhodiola atuntsuensis, Rhodiola himalensis, Rhodiola phariensis, Rhodiola henryi, Rhodiola quadrifida, Rhodiola yunnanensis, Rhodiola wallichiana var. cholaensis, Rhodiola fastigiata, Rhodiola Crassulaceae, Radix Rhodiolae, Rhodiola dumulosa, Rhodiola sachalinensis.
7. the Radix Rhodiolae effective site that each preparation method obtains among the claim 1-6.
8. pharmaceutical composition, it comprises Radix Rhodiolae effective site claimed in claim 7 and pharmaceutically acceptable carrier.
9. pharmaceutical composition according to claim 9 is characterized in that, the dosage form of described pharmaceutical composition can be tablet, capsule, pill, granule, oral liquid or suspensoid.
Radix Rhodiolae effective site preparation treat and/or prevent fatty liver medicine and or health product in application.
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Cited By (7)
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| CN104116774A (en) * | 2014-08-04 | 2014-10-29 | 毕宏生 | Rhodiola rosea extract, extraction method as well as application and product thereof in treatment/prevention of eye disease |
| CN105193898A (en) * | 2015-10-30 | 2015-12-30 | 中国科学院昆明植物研究所 | Rhodiola kirilowii(Regel)Maxim extract and application thereof in cosmetics |
| CN106074667A (en) * | 2016-07-30 | 2016-11-09 | 成都薇诺娜生物科技有限公司 | A kind of method using Radix Rhodiolae effective site preparation treatment fatty liver medicament |
| CN106333985A (en) * | 2016-07-30 | 2017-01-18 | 成都薇诺娜生物科技有限公司 | Method for preparing Rhodiola rosea effective part |
| CN106511437A (en) * | 2016-12-27 | 2017-03-22 | 青海大学 | Preparation method and pharmaceutical application of effective part of rhodiola rosea |
| CN109663123A (en) * | 2018-11-29 | 2019-04-23 | 广州白云山明兴制药有限公司 | Capsule for improving liver injury and body fatigue and preparation method thereof |
| TWI839286B (en) * | 2023-08-08 | 2024-04-11 | 永芙生醫研究有限公司 | Rhodiola rosea nanoemulsion and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104116774A (en) * | 2014-08-04 | 2014-10-29 | 毕宏生 | Rhodiola rosea extract, extraction method as well as application and product thereof in treatment/prevention of eye disease |
| CN105193898A (en) * | 2015-10-30 | 2015-12-30 | 中国科学院昆明植物研究所 | Rhodiola kirilowii(Regel)Maxim extract and application thereof in cosmetics |
| CN106074667A (en) * | 2016-07-30 | 2016-11-09 | 成都薇诺娜生物科技有限公司 | A kind of method using Radix Rhodiolae effective site preparation treatment fatty liver medicament |
| CN106333985A (en) * | 2016-07-30 | 2017-01-18 | 成都薇诺娜生物科技有限公司 | Method for preparing Rhodiola rosea effective part |
| CN106511437A (en) * | 2016-12-27 | 2017-03-22 | 青海大学 | Preparation method and pharmaceutical application of effective part of rhodiola rosea |
| CN109663123A (en) * | 2018-11-29 | 2019-04-23 | 广州白云山明兴制药有限公司 | Capsule for improving liver injury and body fatigue and preparation method thereof |
| CN109663123B (en) * | 2018-11-29 | 2022-05-13 | 广州白云山明兴制药有限公司 | Capsule for improving liver injury and body fatigue and preparation method thereof |
| TWI839286B (en) * | 2023-08-08 | 2024-04-11 | 永芙生醫研究有限公司 | Rhodiola rosea nanoemulsion and preparation method thereof |
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